BIOSSENSOR AND METHOD TO PRODUCE A BIOSSENSOR

专利摘要:
BIOSSENSORS FOR CHEMICAL OR BIOLOGICAL ANALYSIS AND PRODUCTION PROCESSES OF THE SAME A bioseptor, which includes a device base having a set of sensors and a guide arrangement of light guides. The light guides have input regions, which are configured to receive excitation light and light emissions generated by biological or chemical substances. The light guides extend towards the base of the device in the direction of corresponding light sensors and have a filtering material. The base of the device includes a set of circuits, electrically coupled to the light sensors and configured to transmit data signals. The biosensor also includes a shield layer having openings, which are positioned relative to the entry regions of the corresponding light guides, so that light emissions propagate through the openings to the corresponding entry regions. The shield layer extends between the adjacent openings and is configured to block the excitation light and light emissions incident on the shield layer between the adjacent openings.
公开号:BR112016013353B1
申请号:R112016013353-6
申请日:2014-12-09
公开日:2020-11-24
发明作者:Cheng Frank Zhong；Hod Finkelstein；Boyan Boyanov；Dietrich Dehlinger；Darren Segale
申请人:Illumina, Inc；
IPC主号:

专利说明:

RECIPROCAL REMISSION TO RELATED PATENT APPLICATIONS
[001] This patent application claims the benefit and priority of provisional patent application No. 61 / 914.275, filed on December 10, 2013, and bearing the same title, which is incorporated by reference in its entirety in the present descriptive report. BACKGROUND
[002] Embodiments of the present invention generally refer to biological or chemical analysis, and, more particularly, to systems and methods of using detector devices for chemical or biological analysis.
[003] Various protocols in biological or chemical research involve the execution of a large number of controlled reactions on local support surfaces or within predefined reaction chambers. The desired reactions can then be observed or detected, and subsequent analysis can help to identify or reveal the properties of chemicals involved in the reaction. For example, in some multiplex tests, an unknown analyte having an identifiable marker (for example, a fluorescent marker) can be exposed to thousands of known probes under controlled conditions. Each known probe can be deposited in a corresponding receptacle on a microplate. Observation of any chemical reactions, which occur between known probes and the unknown analyte inside the receptacles, can help to identify or reveal analyte properties. Other examples of such protocols include DNA sequencing processes, such as synthesis sequencing (SBS) or cyclic pool sequencing.
[004] In some conventional fluorescence detection protocols, an optical system is used to direct an excitation light in the fluorescence-labeled analyzed, and also to detect the fluorescent signals that can be emitted from the analyzed. However, these optical systems can be relatively expensive and require a larger bench top print. For example, the optical system may include an array of lenses, filters and light sources. In other proposed detection systems, the controlled reactions take place immediately in an image-forming element in the solid state (for example, a charge-coupled device - CCD or a complementary metal oxide semiconductor - CMOS detector), which does not need a large optical array to detect fluorescent emissions.
[005] However, the proposed solid-state imaging systems may have some limitations. For example, it can be challenging to distinguish fluorescent emissions from excitation light when the excitation light is also directed towards the light sensors of the solid-forming image-forming element. In addition, the reagents released in fluid form to those analyzed, which are located on an electronic device, and in a controlled manner, can present additional challenges. As another example, fluorescent emissions are substantially isotropic. As the density of the analyzed in the image-forming element in the solid state increases, it becomes increasingly challenging to control or consider the undesirable light emissions of adjacent analytes (for example, interference). BRIEF DESCRIPTION
[006] In one embodiment, a biosensor is provided, which includes a flow cell and a detector device having the flow cell coupled to it. The flow cell and the detector device form a flow channel, which is configured to contain biological or chemical substances in it, which generate light emissions in response to an excitation light. The detector device includes a device base, having a set of light sensor sensors and a guide arrangement of light guides. The light guides have entrance regions, which are configurations to receive the excitation light and the light emissions from the flow channel. The light guides extend towards the device base, from the input regions towards the light sensors, and have a filtering material, which is configured to filter the excitation light and allow light emissions to propagate towards the light sensors. The device base includes a set of device circuits, electrically coupled to the light sensors and configured to transmit data signals based on photon detectors by the light sensors. The detector device also includes a shield layer, which extends between the flow channel and the device base. The shielding layer has openings, which are positioned relative to the entry regions of corresponding light guides, so that the light emissions propagate through the openings in the corresponding entry regions. The shield layer extends between the adjacent openings, and is configured to block the excitation light and light emissions incident on the shield layer between the adjacent openings.
[007] In one embodiment, a biosensor is provided, which includes a flow cell and a detector device having the flow cell coupled to it. The flow cell and the detector device form a flow channel, which is configured to have biological or chemical substances in it, which generate light emissions in response to an excitation light. The detector device can include a device base, having a set of light sensor sensors and a guide arrangement of light guides. The light guides are configured to receive the excitation light and the light emissions from the flow channel. Each light guide extends to the device base along a central longitudinal axis, from a light guide entry region towards a corresponding light sensor from the sensor set. The light guides include a filter material, which is configured to filter the excitation light and allow light emissions to propagate through it towards the corresponding light sensors. The device base includes a set of device circuits, which are electrically coupled to the light sensors and configured to transmit data signals based on photons detected by the light sensors. The device base includes peripheral interference shields on it, which surround the corresponding light guides of the guide arrangement. The interference shields, at least partially, surround the corresponding light guides around the respective longitudinal axis, to reduce the optical interference between the adjacent light sensors.
[008] In one embodiment, a manufacturing process for a biosensor is provided. The process includes: providing a device base, having a set of light sensor sensors, and a set of device circuits, which are electrically coupled to the light sensors and configured to transmit photon-based data signals detected by the light sensors. light. The device base has an outer surface. The process also includes the application of a shield layer on the outer surface of the device base and the formation of openings by the shield layer. The process also includes the formation of guide cavities, which extend from the corresponding openings towards a corresponding light sensor of the sensor set, and the deposition of filtering material within the guide cavities. A part of the filtering material extends along the shield layer. The process also includes curing the filter material and removing it from the shield layer. The filter material inside the guide cavities forms light guides. The process also includes applying a passivation layer to the shield layer, so that the passivation layer extends directly along the shield layer and through the openings.
[009] In one embodiment, a biosensor is provided, which includes a device base having a set of light sensor sensors and a guide arrangement of light guides. The device base has an outer surface. The light guides have entrance regions, which are configured to receive excitation light and light emissions generated by biological or chemical substances close to the external surface. The light guides extend to the device base, from the input regions towards the corresponding light sensors, and have a filter material, which is configured to filter excitation light and allow light emissions to propagate through it towards the corresponding light sensors. The device base includes a circuitry of the device, which is electrically coupled to the light sensors and configured to transmit data signals based on photons detected by the light sensors. The biosensor also includes a shielding layer, which extends along the outer surface of the device base. The shield layer has openings, which are positioned relative to the entrance regions of the corresponding light guides, so that light emissions propagate through the openings in the corresponding entrance regions. The shield layer extends between the adjacent openings and is configured to block the excitation light and light emissions incident on the shield layer between the adjacent openings.
[010] In one embodiment, a biosensor is provided, which includes a device base having a set of light sensor sensors and a guide arrangement of light guides. The device base has an outer surface. The light guides are configured to receive excitation light and the light emissions generated by biological or chemical substances close to the external surface. Each light guide extends to the device base along a central longitudinal axis, from a light guide entry region towards a corresponding light sensor from the sensor set. The light guide includes a filter material, which is configured to filter the excitation light and allow light emissions to propagate through it towards the corresponding light sensors. The device base includes a circuitry of the device, which is electrically coupled to the light sensors and configured to transmit data signals based on photons detected by the light sensors. The device base includes peripheral interference shields on it, which surround the corresponding light guides of the guide arrangement. The interference shields surround, at least partially, the corresponding light guides around the respective longitudinal axis, to at least block or reflect the errant light rays, to reduce the optical interference between the adjacent light sensors.
[011] Although multiple embodiments are described, still other embodiments of the described object will be evident to those skilled in the technique of detailed description and drawings presented below, which show and describe the illustrative embodiments of the described inventive object. As will be perceived, the inventive object is capable of modifications in several aspects, all of which do not deviate from the spirit and scope of the described object. Consequently, the drawings and the detailed description should be considered as illustrative and not restrictive in nature. BRIEF DESCRIPTION OF THE DRAWINGS
[012] Figure 1 is a block diagram of an exemplary system for biological or chemical analysis, formed according to one embodiment.
[013] Figure 2 is a block diagram of an example system controller, which can be used in the system in Figure 1.
[014] Figure 3 is a block diagram of an exemplary workstation for biological or chemical analysis, according to one embodiment.
[015] Figure 4 is a perspective view of an exemplary workstation and an exemplary cartridge, according to one embodiment.
[016] Figure 5 is a front view of an exemplary shelf set, which includes several workstations in Figure 4.
[017] Figure 6 illustrates the internal components of an exemplary cartridge.
[018] Figure 7 illustrates a cross section of a biosensor, formed according to one embodiment.
[019] Figure 8 is an enlarged part of the cross section of Figure 7, illustrating the biosensor in more detail.
[020] Figure 9 is another enlarged part of the cross section of Figure 7, illustrating the biosensor in more detail.
[021] Figure 10 is a schematic cross section of a detector device, formed according to another embodiment.
[022] Figure 11 is a flow chart illustrating a process for producing a biosensor, according to one embodiment.
[023] Figures 12A and 12B illustrate different stages of manufacture of the biosensor in Figure 11. DETAILED DESCRIPTION
[024] The embodiments described in this specification can be used in various biological or chemical processes and systems for academic or commercial analysis. More specifically, the embodiments described in this specification can be used in various processes and systems in which it is desired to detect an event, a property, a quality or a characteristic, which is indicative of a desired reaction. For example, the embodiments described in this specification include cartridges, biosensors and their components, as well as biological determination systems, which operate with cartridges and biosensors. In particular, the embodiments, cartridges and biosensors include a flow cell and one or more light sensors, which are coupled together in a substantially unitary structure.
[025] Biological determination systems can be configured to perform various desired reactions, which can be detected individually or collectively. Biosensors and biological determination systems can be configured to run several cycles in which the various desired reactions occur in parallel. For example, biological determination systems can be used to sequence a dense set of DNA items through iterative cycles of enzymatic manipulation and image acquisition. In this way, cartridges and biosensors can include one or more fluid microchannels, which transfer reagents or other reaction components to a reaction site. In some embodiments, the reaction sites are randomly distributed over a substantially flat surface. For example, reaction sites may have a non-uniform distribution, in which some reaction sites are located closer together than in other reaction sites. In other embodiments, the reaction sites are patterned over a substantially flat surface in a predetermined manner. Each reaction site can be associated with one or more light sensors, which detect light from the associated reaction site. In still other embodiments, the reaction sites are located in reaction chambers, which compartmentalize the desired reactions in them.
[026] The detailed description presented below of certain embodiments will be better understood when read in conjunction with the attached drawings. Since the figures illustrate diagrams of the functional blocks of various embodiments, the functional blocks are not necessarily indicative of the division between the hardware circuitry. In this way, for example, one or more of the functional blocks (for example, processors or memories) can be implemented in a single piece of hardware (for example, a generic signal processor or a random access memory, a hard disk or the like. ). Similarly, programs can be stand-alone programs, they can be incorporated as subroutines in an operating system, they can be functions in an installed software package, and the like. It should be understood that the various embodiments are not limited to the provisions and instrumentality shown in the drawings.
[027] As used in this specification, an element or step indicated in the singular and preceded with the word "one" or "one" should be understood as not excluding the plural of said elements or steps, unless exclusion is desired explicitly. Furthermore, references to "one embodiment" are not intended to be interpreted as excluding other embodiments, which also incorporate the items indicated. Furthermore, unless explicitly stated otherwise, embodiments "comprising" or "having" an element or elements, having a particular property, may include other elements having or not having that property.
[028] As used in this specification, a "desired reaction" includes a change in at least one of a chemical, electrical, physical or optical property (or quality) of an analysand of interest. In particular embodiments, the desired reaction is a positive binding event (for example, incorporation of a fluorescently labeled biomolecule with the analyte of interest). More generally, the desired reaction can be a chemical transformation, a chemical change or a chemical interaction. The desired reaction can also be a change in electrical properties. For example, the desired reaction may be a change in ionic concentration within a solution. Exemplary reactions include, but are not limited to, chemical reactions, such as reduction, oxidation, addition, elimination, redisposition, esterification, amidation, etherification, cyclization or substitution; binding interactions in which a first chemical binds to a second chemical; dissociation reactions in which two or more chemicals separate from each other; fluorescence; luminescence; bioluminescence; chemiluminescence; and biological reactions, such as nucleic acid replication, nucleic acid amplification, nucleic acid hybridization, nucleic acid binding, phosphorylation, enzymatic catalysis, receptor binding or ligand binding. The desired reaction may be the addition or elimination of a proton, for example, detectable as a change in pH of a solution or surrounding physical medium. An additional desired reaction may be the detection of ion flow through a membrane (for example, natural or synthetic bilayer membrane), for example, as the ions flow through a membrane, the current is interrupted and the interruption can be detected .
[029] In particular embodiments, the desired reaction includes incorporating a fluorescently labeled molecule into an analyte. The analyzed can be an oligonucleotide and the fluorescently labeled molecule can be a nucleotide. The desired reaction can be detected when an excitation light is directed towards an oligonucleotide having the nucleotide labeled, and the fluorophor emits a detectable fluorescent signal. In alternative embodiments, the detected fluorescence is a result of chemiluminescence or bioluminescence. A desired reaction can also increase energy transfer by fluorescence resonance (or Forster) (FRET), for example, by placing a donor fluorophore close to an acceptor fluorophore, decreasing FRET by separating donor fluorophores and acceptor, increase fluorescence by separating a quenching agent from a fluorophore, or decrease fluorescence by colocalizing a quenching agent and a fluorophore.
[030] As used in this specification, a "reaction component" or a "reagent" includes any substance, which can be used to obtain a desired reaction. For example, the reaction components include reagents, enzymes, samples, other biomolecules and buffer solutions. Reaction components are typically transferred to a reaction site in a solution and / or immobilized at a reaction site. The reaction components can interact, directly or indirectly, with another substance, such as the analyzed one of interest.
[031] As used in this specification, the term "reaction site" is a localized region in which a desired reaction can occur. A reaction site can include substrate support surfaces, on which a substance can be immobilized. For example, a reaction site may include a substantially flat surface in a flow cell channel, which has a colony of nucleic acids in it. Typically, but not always, nucleic acids in the colony have the same sequence, being, for example, clonal species of a single or double stranded model. However, in some embodiments, a reaction site may contain only a single nucleic acid molecule, for example, in a single or double stranded form. In addition, several reaction sites can be randomly distributed along the support surface or arranged in a predetermined manner (for example, side by side in a matrix, such as in microwells). A reaction site can also include a reaction chamber, which defines, at least partially, a region or a spatial volume, configured to compartmentalize the desired reaction. As used in this specification, the term "reaction chamber" includes a spatial region, which is in fluid communication with a flow channel. The reaction chamber can be at least partially separated from the physical medium or other surrounding space regions. For example, several reaction chambers can be separated from each other by shared walls. As a more specific example, the reaction chamber can include a cavity, defined by the internal surfaces of a receptacle and have an opening or passage, so that the cavity can be in fluid communication with a flow channel. Biosensors including these reaction chambers are described in more detail in the international patent application No. PCT / US2011 / 057111, filed on October 20, 2011, which is incorporated by reference in its entirety in this specification.
[032] In some embodiments, the reaction chambers are dimensioned and formed relative to the solids (including semi-solids), so that the solids can be inserted, entirely or partially, in them. For example, the reaction chamber can be sized and formed to accommodate only one capture account. The capture account may have clonally amplified DNA or other substances in it. Alternatively, the reaction chamber can be sized and formed to receive an approximate number of beads or solid substrates. As another example, the reaction chambers can also be filled with a gel or a porous substance, which is configured to control diffusion or filter fluids that can flow into the reaction chamber.
[033] In some embodiments, light sensors (for example, photodiodes) are associated with the corresponding reaction sites. A light sensor, which is associated with a reaction site, is configured to detect light emissions from the associated reaction site, when a desired reaction has occurred at the associated reaction site. In some cases, multiple light sensors (for example, multiple pixels from a camera device) can be associated with a single reaction site. In other cases, a single light sensor (for example, a single pixel) can be associated with a single reaction site or with a group of reaction sites. The light sensor, the reaction site and other items of the biosensor can be configured so that at least part of the light is detected directly by the light sensor, without being reflected.
[034] As used in this specification, the term "adjacent", when used in relation to two reaction sites, means that no other reaction site is located between the two reaction sites. The term "adjacent" can have a similar meaning when used with respect to adjacent detection routes and adjacent light sensors (for example, adjacent light sensors that have no other light sensors between them). In some cases, a reaction site may not be adjacent to another reaction site, but it may still be within an immediate vicinity of the other reaction site. A first reaction site may be in the immediate vicinity of a second reaction site, when fluorescent emission signals from the first reaction site are detected by the light sensor associated with the second reaction site. More specifically, a first excitation light can be in the immediate vicinity of a second excitation light, when the light sensor, associated with the second reaction site, detects, for example, interference from the first reaction site. Adjacent reaction sites may be contiguous so that they come in contact with each other, or adjacent sites may not be contiguous having an intermediate space between them.
[035] As used in this specification, a "substance" includes items or solids, such as branch capture accounts, as well as biological or chemical substances. As used in this specification, a "biological or chemical substance" includes biomolecules, samples of interest, analytes of interest and other or other chemical compounds. A biological or chemical substance can be used to detect, identify or analyze another or other chemical compounds, or it works as intermediates to study or analyze another or other chemical compounds. In particular embodiments, chemical or biological substances include a biomolecule. As used in this specification, a "biomolecule" includes at least one biopolymer, a nucleotide, a nucleic acid, a polynucleotide, an oligonucleotide, a protein, an enzyme, a polypeptide, an antibody, an antigen, an ligand, a receptor, a polysaccharide, a carbohydrate, a polyphosphate, a cell, a tissue, an organism or a fragment thereof, or any other or any other biologically active chemical compounds, such as analogues or replicates of the species mentioned above.
[036] In another example, a biological or chemical substance or a biomolecule includes an enzyme or a reagent, used in a coupled reaction to detect the product of another reaction, such as an enzyme or a reagent, used to detect pyrophosphate in a pyro-sequencing reaction. Enzymes and reagents, useful for the detection of pyrophosphate, are described, for example, in the publication of U.S. patent application No. 2005/0244870 A1, which is incorporated in the present specification in its entirety.
[037] Biomolecules, samples and biological or chemical substances can be natural or synthetic, and can be suspended in a solution or mixture within a space region. Biomolecules, samples or chemicals can also be attached to a solid phase or a gel material. Biomolecules, samples and biological or chemical substances can also include a pharmaceutical composition. In some cases, biomolecules, samples and biological or chemical substances of interest can be referred to as targets, probes or analyzed.
[038] As used in this specification, a "biosensor" includes a structure having multiple reaction sites, which is configured to detect desired reactions, occurring at or near the reaction sites. A biosensor can include a solid state image forming device (e.g., CCD or CMOS image forming device) and, optionally, a flow cell mounted thereon. The flow cell can include at least one flow channel, which is in fluid communication with the reaction sites. As a specific example, the biosensor is configured to fluidly and electrically couple with a bioassay system. The bioassay system can distribute the reagents to the reaction sites, according to a predetermined protocol (for example, sequencing by synthesis), and perform various imaging events. For example, the bioassay system can direct solutions to flow through the reaction sites. At least one of the solutions can include four types of nucleotides, having the same or different fluorescent markers. Nucleotides can bind to the corresponding oligonucleotides, located at the reaction sites. The bioassay system can then illuminate the flow channels using an excitation light source (for example, solid-state light sources, such as light-emitting diodes or LEDs). The excitation light can have one or more predetermined wavelengths, including a range of wavelengths. The excited fluorescent markers provide emission signals, which can be detected by light sensors.
[039] In alternative embodiments, the biosensor may include electrodes or other types of sensors, configured to detect other identifiable properties. For example, sensors can be configured to detect a change in ionic concentration. In another example, sensors can be configured to detect the flow of ionic current through a membrane.
[040] As used in this specification, a "cartridge" includes a structure, which is configured to retain a biosensor. In some embodiments, the cartridge may include other items, such as the light source (e.g., LEDs), that are configured to provide excitation light to the reaction sites of the biosensor. The cartridge can also include a fluid storage system (eg, storage for reagents, sample and buffer) and a control system for fluid (eg, pumps, valves and the like) to fluidly transport the reaction components, the sample and similar to the reaction sites. For example, after the biosensor is prepared or produced, the biosensor can be coupled to a housing or container on the cartridge. In some embodiments, biosensors and cartridges can be self-contained, disposable units. However, other embodiments may include a set with removable parts, which allow a user to have access to an internal part of the biosensor or cartridge, for maintenance or replacement of components or samples. The biosensor and the cartridge can be attached or removed removably to systems of larger biological determinations, such as a sequencing system, which conduct the controlled reactions in it.
[041] As used in this specification, when the terms "removably" and "coupled" (or "stuck") are used together to describe a relationship between the biosensor (or cartridge) and a receptacle in a system or a interface of a bioassay system, the term is intended to mean that a connection between the biosensor (or the cartridge) and the system receptacle is easily separable, without destroying or damaging the system receptacle and / or the biosensor (or cartridge) . Components are easily separable when components can be separated from each other without undue effort or a significant amount of time spent separating the components. For example, the biosensor (or the cartridge) can be attached or removably attached to the system receptacle electrically, so that the joint, which comes in contact with the bioassay system, is not destroyed or damaged. The biosensor (or the cartridge) can also be mechanically attached or removably attached, so that items that retain the biosensor (or the cartridge) are not destroyed or damaged. The bi-sensor (or the cartridge) can also be attached or removably attached to the system receptacle fluidly, so that the holes in the system receptacle are not destroyed or damaged. The receptacle or a component of the system is not considered to be destroyed or damaged if, for example, only a simple adjustment to the component (for example, realignment) or a simple replacement (for example, replacement of a nozzle) is necessary.
[042] As used in this specification, the term "fluid communication" or "fluidly coupled" refers to two spatial regions being connected together, so that a liquid or gas can flow between the two spatial regions. For example, a fluid microchannel can be in fluid communication with a reaction chamber, so that a fluid can flow freely into the reaction chamber of the fluid microchannel. The term "in fluid communication" or "fluidly coupled" also indicates that two spatial regions, in fluid communication by one or more valves, limiters or other components for fluids, are configured to control or regulate a flow of fluid through a system.
[043] As used in this specification, the term "immobilized", when used in connection with a biomolecule or biological or chemical substance, substantially includes the attachment of the biomolecule or biological or chemical substance to a molecular level on a surface. For example, a biomolecule or a biological or chemical substance can be immobilized on a surface of the substrate material, using absorption techniques including non-covalent interactions (for example, electrostatic forces, van der Waals forces and dehydration of interfaces hydrophobic), and covalent bonding techniques, in which functional groups or bonding agents facilitate the attachment of biomolecules on the surface. The immobilization of biomolecules or biological or chemical substances on a surface of a substrate material can be based on the surface properties of the substrate, in the liquid medium leading to biomolecule or biological or chemical substance, and on the properties of the biomolecules or biological substances themselves or chemical. In some cases, a surface of the substrate can be functionalized (for example, chemically or physically modified), to facilitate the immobilization of biomolecules (or biological or chemical substances) on the surface of the substrate. The substrate surface can first be modified to have functional groups attached to the surface. Functional groups can then attach to biomolecules or biological or chemical substances, to immobilize them in them. A substance can be immobilized on a surface by means of a gel, for example, as described in the publication of U.S. patent application No. 2011/0059865 A1, which is incorporated into this specification by reference.
[044] In some embodiments, nucleic acids can be trapped on a surface and amplified using bridge amplification. Useful bridge amplification processes are described, for example, in US patent 5,641,658, international patent application WO 07/010251, US patent 6,090,592, publication of US patent application No. 2002/005510 A1, patent US 7,115,400, publication of US patent application No. 2004/0096853 A1, publication of US patent application No. 2004/0002090 A1, publication of US patent application No. 2007/0128624 A1 and publication of application US Patent No. 2008/0009420 A1, all of which are incorporated in their entirety in this specification. Another useful process for amplifying nucleic acids on a surface is rolling circle amplification (RCA), for example, using the processes presented in more detail below. In some embodiments, nucleic acids can be trapped on a surface and amplified using one or more pairs of primers. For example, one of the primers can stay in solution and the other primer can be immobilized on the surface (for example, stuck in position 5 '). By way of example, a nucleic acid molecule can hybridize one of the primers on the surface, followed by the extension of the immobilized primer, to produce a first copy of the nucleic acid. The solution primer then hybridizes to the first copy of the nucleic acid, which can be extended by using the first copy of the nucleic acid, as a template. Optionally, after the first copy of the nucleic acid is produced, the original nucleic acid molecule can hybridize to a second primer immobilized on the surface and can be extended at the same time or after extension of the primer in solution. In any embodiment, repeated rounds of extension (e.g., amplification), using the immobilized primer and the solution primer, provide multiple copies of the nucleic acid.
[045] In particular embodiments, the determination protocols, performed by the systems, and the processes, described in this specification, include the use of natural nucleotides and also enzymes, which are configured to interact with the natural nucleotides. Natural nucleotides include, for example, ribonucleotides or deoxyribonucleotides. The natural nucleotides can be in the form of mono-, di- or triphosphate and can have a selected base of adenine (A), thymine (T), uracil (U), guanine (G) or cytosine (C). It should be understood, however, that unnatural nucleotides, modified nucleotides or nucleotide analogues mentioned above can be used. Some examples of useful unnatural nucleotides are presented below in relation to sequencing based on reversible terminator by the synthesis processes.
[046] In embodiments that include the reaction chambers, items or solid substances (including semi-solid substances) can be disposed within the reaction chambers. When disposed, the item or solid can be physically retained or immobilized within the reaction chamber through an interference, adhesion or retention fit. Exemplary items or solids, which can be disposed within the reaction chambers, include polymer beads, pellets, agarose gel, powders, quantum dots or other solids, which can be compressed and / or retained within the reaction chamber. In particular embodiments, a nucleic acid superstructure, such as a DNA sphere, can be arranged in or in a reaction chamber, for example, by attachment to an internal surface of the reaction chamber, or by residence in a liquid within the reaction chamber. reaction chamber. One sphere of DNA or the other nucleic acid superstructure can be preformed and then disposed in or in a reaction chamber. Alternatively, a sphere of DNA can be synthesized in situ in the reaction chamber. A DNA sphere can be synthesized by rolling circle amplification to produce a concatamer of a particular nucleic acid sequence, and the concatamer can be treated under conditions of forming a relatively compact sphere. The DNA spheres and the processes for their synthesis are described, for example, in U.S. patent publications numbers 2008/0242560 A1 and 2008/0234136 A1, both of which are incorporated in the present specification in their entirety. A substance, which is retained or disposed of in a reaction chamber, can be in a solid, liquid or gaseous state.
[047] Figure 1 is a block diagram of an exemplary bioassay system 100, for biological or chemical analysis, formed according to one embodiment. The term "bioassay system" is not intended to be limiting, as the bioassay system 100 can operate any information or data, which refers to at least one of a biological or chemical substance. In some embodiments, the bioassay system 100 is a workstation, which can be similar to a bench top device or a desktop computer. For example, a majority (or all) of the systems and components, to conduct the desired reactions, can be contained within a common housing.
[048] In particular embodiments, the bioassay system 100 is a nucleic acid sequencing (or sequencing) system, configured for a variety of applications, including, but not limited to, new sequencing, a resequencing of all genomes or genomic regions of interest, and meta-genomics. The sequencer can also be used for DNA or RNA analysis. In some embodiments, the bioassay system 100 can also be configured to generate reaction sites on a biosensor. For example, the bioassay system 100 can be configured to receive a sample and generate clusters trapped on the surface of clonal amplified nucleic acids derived from the sample. Each cluster can constitute or be part of a reaction site in the biosensor.
[049] The exemplary bioassay system 100 can include a receptacle or an interface of system 112, which is configured to interact with a biosensor 102, to perform the desired reactions within biosensor 102. In the description presented below with respect to Figure 1 , biosensor 102 is placed in the system 112 receptacle. However, it should be understood that a cartridge, which includes biosensor 102, can be inserted into the system 112 receptacle, and, in some states, the cartridge can be removed temporarily or permanently. As described above, the cartridge can include, among other things, fluid control and fluid storage components.
[050] In particular embodiments, the bioassay system 100 is configured to perform a large number of parallel reactions within biosensor 102. Biosensor 102 includes one or more reaction sites, in which the desired reactions can occur. The reaction sites can be, for example, immobilized on a solid surface of the biosensor or immobilized on beads (or other mobile substrates), which are located inside the corresponding reaction chambers of the biosensor. Reaction sites can include, for example, clonally amplified nucleic acid clusters. Biosensor 102 can include a solid state image forming device (e.g., a CCD or CMOS image forming device) and a flow cell mounted thereon. The flow cell can include one or more flow channels, which receive a solution from the bioassay system 100 and direct the solution towards the reaction sites. Optionally, biosensor 102 can be configured to couple a thermal element, to transfer thermal energy to and from the flow channel.
[051] The bioassay system 100 may include several components, assemblies and systems (or subsystems), which interact to conduct a predetermined test process or protocol for biological or chemical analysis. For example, the bioassay system 100 includes a system controller 104, which can communicate with the various components, assemblies and subsystems of the bioassay system 100 and also of biosensor 102. For example, in addition to the system receptacle 112, the system Bioassay 100 may also include: a fluid control system 106, for controlling fluid flow through a fluid network of the bioassay system 100 and biosensor 102; a fluid storage system 108, which is configured to hold all fluids (for example, gases or liquids), which can be used by the bioassay system; a temperature control system 110, which can regulate the fluid temperature in the fluid network, in the fluid storage system 108 and / or in the biosensor 102; and a lighting system 111, which is configured to illuminate biosensor 102. As described above, if a cartridge, having biosensor 102, is placed in the 112 system receptacle, the cartridge can also include fluid control and storage components of fluid.
[052] Also shown is the bioassay system 100, which may include a user interface 114, which interacts with the user. For example, user interface 114 may include a monitor 113, for displaying or requesting information from a user, and a user input device 115, for receiving user input. For example, user interface 114 may include a touch screen, configured to detect the presence of a person's touch and also identify a touch location on the screen. However, other 115 user input devices can be used, such as a mouse, a touch table, a keyboard, a compact keyboard, a hand scanner, a voice recognition system, a motion recognition system and similar. As will be discussed in more detail below, the bioassay system 100 can communicate with various components, including biosensor 102 (for example, in the form of a cartridge), to perform the desired reactions. The bioassay system 100 can also be configured to analyze the data obtained from the biosensor, to provide a user with desired information.
[053] System controller 104 can include any processor or microprocessor based system, including systems using microcontrollers, reduced instruction set computers (RISC), application specific integrated circuits (ASICs), a set of programmable ports on the field (FGPAs), logic circuits, and any other circuit or processor capable of performing the functions described in this specification. The examples presented above are exemplary only, and are therefore not intended to limit the definition and / or the meaning of the term system controller in any way. In the exemplary embodiment, the system controller 104 executes a set of instructions, which is stored in one or more storage elements, memories or modules, for at least one of obtaining and analyzing the detection data. The storage elements can be in the form of information sources or physical memory elements within the bioassay system 100.
[054] The instruction set can include several commands, which instruct the bioassay system 100 or the biosensor 102, to perform specific operations, such as the processes and methods of the various embodiments described in this specification. The set of instructions can be in the form of a software program, which can form part of a non-transitory, tangible, computer-readable medium or medium. As used in this specification, the terms "software" and "hardware programming" are interchangeable, and include any computer program stored in memory, for execution by a computer, including RAM, ROM, EPROM, an EEPROM memory and a non-volatile RAM memory (NVRAM). The types of memories mentioned above are only exemplary, and are thus not limiting the types of memory useful for storing a computer program.
[055] The software can be in various forms, such as system software or application software. Furthermore, the software can be in the form of a set of separate programs, or a program module within a larger program or a part of a program module. The software can also include modular programming in the form of object-oriented programming. After obtaining the detection data, the detection data can be automatically processed by the bioassay system 100, processed in response to user input, or processed in response to a request made by another processing machine (for example, a remote request by communication link).
[056] System controller 104 can be connected to biosensor 102 and other components of the bioassay system 100 via communication links. System controller 104 can also be communicatively connected to off-site systems or servers. Communication links can be connected or wireless. System controller 104 can receive user inputs or commands from user interface 114 and user input device 115.
[057] Fluid control system 106 includes a fluid network and is configured to direct and regulate the flow of one or more fluids through the fluid network. The fluid network can be in fluid communication with the biosensor 102 and the fluid storage system 108. For example, the selected fluids can be taken from the fluid storage system 108 and directed in a controlled manner to the biosensor 102, or the fluids can be removed from biosensor 102 and directed, for example, towards a disposal reservoir in fluid storage system 108. Although not shown, fluid control system 106 may include flow sensors, which detect a flow or pressure of fluids within the fluid network. The sensors can communicate with the system controller 104.
[058] The temperature control system 110 is configured to regulate the temperature of fluids in different regions of the fluid network, the fluid storage system and / or the biosensor 102. For example, the temperature control system 110 can include a thermal cycling device, which interfaces with biosensor 102 and controls the temperature of the fluid, which flows along the reaction sites in biosensor 102. Temperature control system 110 can also regulate the temperature of solid elements or components of the bioassay system 100 or biosensor 102. Although not shown, the temperature control system 110 may include sensors, to detect the temperature of the fluid or other components. The sensors can communicate with the system controller 104.
[059] Fluid storage system 108 is in fluid communication with biosensor 102 and can store various reaction components or reagents, which are used to conduct the desired reactions in it. Fluid storage system 108 can also store fluids for flushing or cleaning the fluid network and biosensor 102 and for diluting reagents. For example, fluid storage system 108 may include various reservoirs for storing samples, reagents, enzymes, other biomolecules, buffer solutions, aqueous and non-polar solutions, and the like. In addition, fluid storage system 108 can also include disposal reservoirs to receive disposal products from biosensor 102. In embodiments that include a cartridge, the cartridge can include one or more of a fluid storage system, a fluid control system or a temperature control system. Consequently, one or more of the components presented in this specification as associated with these systems can be contained within a cartridge housing. For example, a cartridge can have several reservoirs to store samples, reagents, enzymes, other biomolecules, buffer solutions, aqueous and non-polar solutions, disposal, and the like. As such, one or more of a fluid storage system, a fluid control system or a temperature control system can be removably attached to a bioassay system by means of a cartridge or other biosensor.
[060] Lighting system 111 may include a light source (for example, one or more LEDs) and several optical components, to illuminate the biosensor. Examples of light sources can include lasers, arc lamps, LEDs, or laser diodes. Optical components can be, for example, reflectors, dichroic components, beam splitters, lenses, filters, wedges, prisms, mirrors, detectors and the like. In embodiments using a lighting system, the lighting system 111 can be configured to direct an excitation light to the reaction sites. As an example, fluorophores can be excited by wavelengths of green light, as this wavelength of the excitation light can be approximately 532 nm.
[061] The receptacle or system interface 112 is configured to hold biosensor 102 in at least one mechanical, electrical and fluid manner. The system receptacle 112 can retain biosensor 102 in a desired orientation, to facilitate fluid flow through biosensor 102. The system receptacle 112 can also include electrical contacts, which are configured for attachment to biosensor 102, so that the system of bioassay 100 can communicate with biosensor 102 and / or provide energy to biosensor 102. In addition, system receptacle 112 may include fluid holes (for example, nozzles), which are configured to attach to biosensor 102. In in some embodiments, biosensor 102 is removably coupled to the system 112 receptacle mechanically, electrically, and also fluidly.
[062] In addition, the bioassay 100 system can communicate remotely with other systems or networks, or with other bioassay 100 systems. The detection data, obtained by the 100 bioassay systems or systems can be stored in a database remote.
[063] Figure 2 is a block diagram of system controller 104 in the exemplary embodiment. In one embodiment, system controller 104 includes one or more processors or modules, which can communicate with each other. All processors or modules can include an algorithm (for example, instructions stored in a non-transitory and / or tangible computer-readable storage medium) or sub-algorithms to execute particular processes. System controller 104 is conceptually illustrated as a set of modules, but can be implemented using any combination of dedicated hardware cards, DSPs, processors, etc. Alternatively, system controller 104 can be implemented using a commercial PC with a single processor or multiple processors, with the functional operations distributed among the processors. As another option, the modules described below can be implemented using a hybrid configuration, in which some modular functions are performed using dedicated hardware, while the remaining modular functions are performed using a commercial PC and the like. The modules can also be implemented as software modules within a processing unit.
[064] During operation, a communication link 120 can transmit information (for example, commands) to, or receive information (for example, data) from, biosensor 102 (Figure 1) and / or subsystems 106, 108, 110 (Figure 1). A communication link 122 can receive user input from user interface 114 (Figure 1) and transmit data or information to user interface 114. Data from biosensor 102 or from subsystems 106, 108, 110 can be processed by the system controller 104 in real time, during a bioassay session. Additionally or alternatively, the data can be temporarily stored in a system memory, during a bioassay session, and processed in a slower operation than in real time or offline.
[065] As shown in Figure 2, system controller 104 can include several modules 131 - 139, which communicate with a main control module 130. Main control module 130 can communicate with user interface 114 (Figure 1). Although modules 131-139 are shown as in direct communication with main control module 130, modules 131-139 can also communicate directly with each other, with the sulfur interface 114 and with biosensor 102. Also, the modules 131-139 can communicate with the main control module 130 through the other modules.
[066] The various modules 131-139 include system modules 131-133, 139, which communicate with subsystems 106, 108, 110 and 111, respectively. The fluid control module 131 can communicate with the fluid control system 106, to control the valves and flow sensors of the fluid network, to control the flow of one or more fluids through the fluid network. The fluid storage module 132 can notify the user when the fluid level is low, or when the discharge reservoir is at or near its capacity. The fluid storage module 132 can also communicate with the temperature control module 133, so that fluids can be stored at a desired temperature. The lighting module 139 can communicate with the lighting system 109, to illuminate the reaction sites at the desired times, during a protocol, such as after the desired reactions have occurred (e.g., switching events).
[067] The various modules 131-139 may also include a device module 134, which communicates with biosensor 102, and an identification module 135, which determines identification information relating to biosensor 102. Device module 134 can , for example, communicating with the system receptacle 112, to confirm that the biosensor has established an electrical and fluid connection with the bioassay system 100. Identification module 135 can receive signals, which identify biosensor 102. The identification module 135 can use the identity of biosensor 102 to provide other information to the user. For example, identification module 135 can determine and then display a batch number, a production date, or a protocol that is recommended to be run with biosensor 102.
[068] The various modules 131-139 can also include a detection data analysis module 138, which receives and analyzes the signal data (for example, image data) from biosensor 102. The signal data can be stored for subsequent analysis, or can be transmitted to user interface 114, to display the desired information to the user. In some embodiments, the signal data can be processed by the solid state image forming device (e.g., the CMOS image sensor), before the detection data analysis module 138 receives the signal data.
[069] Protocol modules 136 and 137 communicate with main control module 130, to control the operation of subsystems 106, 108 and 110, when conducting predetermined test protocols. Protocol modules 136 and 137 can include sets of instructions to instruct the bioassay system 100 to perform specific operations associated with predetermined protocols. As shown, the protocol module can be a synthesis sequencing module (SBS) 136, which is configured to issue various commands for executing synthesis sequencing processes. In SBS, the extent of a nucleic acid primer across a nucleic acid model is monitored to determine the nucleotide sequence in the model. The associated chemical process can be polymerization (for example, catalyzed by a polymerase enzyme) or ligation (for example, catalyzed by a ligase enzyme). In an embodiment of polymerase-based SBS, fluorescently labeled nucleotides are added to a primer (thereby extending the primer) in a model-dependent mode, so that detection of the order and type of nucleotides added to the primer can be used to determine the model sequence. For example, to initiate a first cycle of SBS, commands can be provided to transfer one or more labeled nucleotides, DNA polymerase, etc. to attach a flow cell, which houses a set of nucleic acid models. The nucleic acid models can be located at corresponding reaction sites. These reaction sites, in which the extension of the primer causes a labeled nucleotide to be incorporated, can be detected by an image formation event. During an imaging event, the lighting system 111 can provide an excitation light to the reaction sites. Optionally, the nucleotides can further include a reversible termination property, which terminates another extension of primer, once the nucleotide has been added to a primer. For example, a nucleotide analog, having a reversible terminator part, can be added to a primer, so that subsequent extension cannot occur until an unlocking agent is released to remove the part. Thus, for embodiments using reversible termination, a command can be provided to release an unlocking reagent to the flow cell (before or after detection has occurred). One or more commands can be provided to promote one or more washes between the various release steps. The cycle can then be repeated n times, to extend the primer by n nucleotides, thereby detecting a sequence of length n. Exemplary sequencing techniques are described, for example, by Bentley et al., Nature 456: 53 - 59 (2008), WO 04/018497, US 7,057,026, WO 91/06678, WO 07/123744, US 7,329. 492, US 7,211,414, US 7,315,019, US 7,405,281 and US 2008/0108082, all of which are incorporated by reference in this specification.
[070] For the nucleotide release step of an SBS cycle, a single type of nucleotide can be released at a time, or multiple different types of nucleotides (for example, A, C, T and G together) can be released. For a nucleotide release configuration, in which only a single type of nucleotide is present at a time, the different nucleotides do not need to have distinct markers, since they can be distinguished based on a temporal separation, inherent in individualized release. Consequently, a sequencing process or apparatus can use a single color detection. For example, an excitation source need only provide excitation at a single wavelength or in a single range of wavelengths. For a nucleotide release configuration, in which the release results in multiple different nucleotides being present in the flow cell at a time, the sites, which incorporate different types of nucleotides, can be distinguished based on different fluorescent markers, which are trapped in the respective types of nucleotides in the mixture. For example, four different nucleotides can be used, each having one of four different fluorophores. In one embodiment, the four different fluorophores can be distinguished using excitation in four different regions of the spectrum. Alternatively, less than four different excitation sources can be used, but optical filtration of excitation radiation, from a single source, can be used to produce different ranges of excitation radiation in the flow cell.
[071] In some embodiments, less than four different colors can be detected in a mixture having four different nucleotides. For example, pairs of nucleotides can be detected at the same wavelength, but distinguished based on a difference in intensity for one element of the pair compared to the other, or based on a change to an element of the pair (e.g. through chemical modification, photochemical modification or physical modification), which causes the clear signal to appear or disappear, compared to the signal detected for the other element of the pair. Exemplary apparatus and processes for distinguishing four different nucleotides, using the detection of less than four colors, are described, for example, in US patent applications 61 / 538,294 and 61 / 619,878, which are incorporated in this report descriptive by reference in its entirety. U.S. patent application No. 13 / 624,200, which was filed on September 21, 2012, is also incorporated by reference in its entirety.
[072] The various protocol modules can also include a sample preparation (or generation) module 137, which is configured to issue commands to the fluid control system 106 and temperature control system 110, to amplify a product within of biosensor 102. For example, biosensor 102 can be coupled to the bioassay system 100. Amplification module 137 can issue instructions to fluid control system 106 to release the necessary amplification components to the reaction chambers inside biosensor 102 In other embodiments, the reaction sites may already contain some components for amplification, such as model DNA and / or primers. After releasing the amplification components to the reaction chambers, the amplification module 137 can instruct the temperature control system 110, to alternate through different temperature stages, according to the known amplification protocols. In some embodiments, the amplification and / or incorporation of the nucleotide is or is done isothermally.
[073] The SBS 136 module can issue commands to perform bridged PCR when clusters of clonal amplicons are formed in areas located within a flow cell channel. After generation of the amplicons by bridged PCR, the amplicons can be "linearized" to produce single stranded model DNA, or sstDNA, and a sequencing primer can be hybridized to a universal sequence, which borders a region of interest . For example, sequencing based on a reversible terminator by a synthesis process can be used as shown above or as follows.
[074] Each sequencing cycle can extend an sstDNA by a single base, which can be done, for example, by using a modified DNA polymerase and a mixture of four types of nucleotides. The different types of nucleotides can have unique fluorescent markers, and each nucleotide can also have a reversible terminator, which allows a single base incorporation to occur in each cycle. After a single base is added to the sstDNA, the excitation light can be incident at the reaction sites and fluorescent emissions can be detected. Upon detection, the fluorescent marker and terminator can be chemically decomposed from the sstDNA. Another similar sequencing cycle can be done next. In this sequencing protocol, module SBS 136 can instruct fluid control system 106 to direct a flow of reagent and enzyme solutions through biosensor 102. Exemplary reversible terminator-based SBS processes that can be used with devices and processes, presented in this specification, are described in the publication of US patent application No. 2007/0166705 A1, US patent 7,057,026, publication of US patent application No. 2006/0240439 A1, publication of US patent application No. 2006/0281109 A1, PCT publication of WO 05/065814, publication of US patent application No. 2005/0100900 A1, PCT publication of WO 06/064199 and PCT publication of WO 07/010251, all of which are incorporated in this specification by reference in their entirety. Exemplary reagents for reversible terminator-based SBS are described in US patents 7,541,444, US 7,057,026, US 7,414,116, US 7,427,673, US 7,566,537, US 7,592,435 and in international patent application WO 07/135368, all of which are incorporated in this specification by reference in their entirety.
[075] In some embodiments, the amplification and SBS modules can operate on a single assay protocol, in which, for example, the model's nucleic acid is amplified and subsequently sequenced within the same cartridge.
[076] The bioassay system 100 can also allow the user to reconfigure a test protocol. For example, the bioassay system 100 may offer options to the user through user interface 114, to modify the given protocol. For example, if it is determined that biosensor 102 is going to be used for amplification, the bioassay system 100 may request a temperature for the annealing cycle. Furthermore, the bioassay system 100 can issue warnings to a user, if that user has provided user inputs, which are not generally acceptable for the selected test protocol.
[077] Figure 3 is a block diagram of an exemplary workstation 200 for biological or chemical analysis, according to one embodiment. Workstation 200 may have items, systems and assemblies similar to the bioassay system 100, described above. For example, workstation 200 may have a fluid control system, such as fluid control system 106 (Figure 1), which is coupled in fluid communication to a biosensor (or cartridge) 235 via a network fluid 238. Fluid network 238 may include a reagent cartridge 240, a valve block 242, a main pump 244, a bubble remover 246, a three-way valve 248, a flow restrictor 250, a flow system discharge removal 252 and a purge pump 254. In particular embodiments, most or all of the components, described above, are housed in a common workstation housing (not shown). Although not shown, workstation 200 may also include an illumination system, such as illumination system 111, which is configured to provide excitation light to the reaction sites.
[078] Fluid flow is indicated by arrows along fluid network 238. For example, reagent solutions can be removed from reagent cartridge 240 and flow through valve block 242. Valve block 242 can facilitate creation of zero dead volume of fluid flowing to cartridge 235 of reagent cartridge 240. Valve block 242 can select or allow one or more liquids within reagent cartridge 240 to flow through fluid network 238. For example, the block valve valve 242 may include solenoid valve, which have a compact arrangement. Each solenoid valve can control the flow of fluid from a single reservoir bag. In some embodiments, valve block 242 may allow two or more different liquids to flow into fluid network 238 at the same time, thereby mixing the two or more different liquids. After leaving the valve block 242, fluid can flow through the main pump 244 and the bubble eliminator 246. The bubble eliminator 246 is configured to remove unwanted gases that have entered or been generated within the fluid network 238.
[079] From the bubble eliminator 246, the fluid can flow to the three-way valve 248, in which the fluid is directed to cartridge 235 or diverted to the disposal system 252. A flow of fluid into cartridge 235 it can be, at least partially, controlled by the flow restrictor 250, located downstream of the cartridge 235. Furthermore, the flow restrictor 250 and the main pump 244 can be coordinated with each other, to control the flow of fluid through the reaction and / or control pressure within fluid network 238. Fluid can flow through cartridge 235 and into the waste removal system 252. Optionally, fluid can flow through purge pump 254 and, for example, a bag disposal tank inside reagent cartridge 240.
[080] Workstation 200 is also shown in Figure 3, which can include a temperature control system, such as the temperature control system 110, which is configured to regulate or control a thermal environment of the different components and workstation 200 subsystems. Temperature control system 110 may include a reagent cooler 264, which is configured to control the temperature requirements of the various fluids used by workstation 200, and a thermal cycling device 266, which is configured to control the temperature of a cartridge 235. The thermal cycling device 266 may include a thermal element (not shown), which interfaces with the cartridge.
[081] Furthermore, workstation 200 may include a system controller or an SBS 260 card, which may be items similar to those on system controller 104 described above. The SBS 260 card can communicate with the various components and subsystems of workstation 200, as well as with the 235 cartridge. In addition, the SBS 260 card can communicate with remote systems, for example, to store data or receive commands from remote systems. Workstation 200 may also include a touch screen user interface 262, which is operationally coupled to card 260 by a single board computer (SBC) 272. Workstation 200 may also include one or more ports and / or accessible data communication drives. For example, a workstation 200 may include one or more universal serial bus (USB) connections to computer peripherals, such as an instantaneous or jump drive, a compact instantaneous (CF) drive and / or a 270 hard drive, to store user data in addition to other software.
[082] Figure 4 is a perspective view of a workstation 300 and a cartridge 302, which may include one or more biosensors (not shown), as described in this specification. Workstation 300 may include components similar to those described with respect to the bioassay system 100 and workstation 200, and may operate in a similar manner. For example, workstation 300 may include workstation housing 304, and a system receptacle 306, which is configured to receive and couple cartridge 302. The system receptacle may be coupled either fluidly or electrically to cartridge 302. Workstation housing 304 can hold, for example, a system controller, a fluid storage system, a fluid control system, and a temperature control system, as described above. In Figure 4, workstation 300 does not include a user interface or screen, which is coupled to workstation housing 304. However, a user interface can be communicatively coupled to housing 304 (and components / systems in it) through a communication link. In this way, the user interface and workstation 300 can be located remotely relative to each other. Together, the user interface and workstation 300 (or several of them) can constitute a bioassay system.
[083] As shown, cartridge 302 includes a cartridge housing 308, having at least one orifice 310, which provides access to an inner portion of the cartridge housing 308. For example, a solution, which is configured to be used in the cartridge 302, during controlled reactions, can be inserted through hole 310 by a technician or through workstation 300. System receptacle 306 and cartridge 302 can be sized and formed relatively to each other, so that cartridge 302 can be inserted in a receptacle cavity (not shown) of the 306 system receptacle.
[084] Figure 5 is a front view of a shelf assembly 312, with a cabinet or cart 314 with several workstations 300 placed on it. Cabinet 314 may include one or more shelves 316, which define one or more reception spaces 318, configured to receive one or more workstations 300. Although not shown, workstations 300 can be coupled in communication to a network of communication, which allows a user to control the operation of 300 workstations. In some embodiments, a bioassay system includes multiple workstations, such as workstations 300, and a single user interface configured to control operation multiple workstations.
[085] Figure 6 illustrates various items from cartridge 302 (Figure 4), according to one embodiment. As shown, cartridge 302 can include a sample set 320, and the receptacle of system 306 can include a light set 322. The decking 346, shown in Figure 6, represents the spatial relationship between the first and second subsets 320 and 322 , when they are separated from each other. At stage 348, the first and second subsets 320 and 322 are joined together. The cartridge housing 308 (Figure 4) can wrap the first and second subsets 320 and 322 joined together.
[086] In the illustrated embodiment, the first subset 320 includes a base 326 and a reaction component body 324, which is mounted on base 326. Although not shown, one or more biosensors can be mounted on base 326 in a recess 328, which is defined, at least in part, by the reaction component body 324 and base 326. For example, at least four biosensors can be mounted on base 326. In some embodiments, base 326 is a printed circuit board having a circuitry, which allows communication between the different components of the cartridge and the workstation 300 (Figure 4). For example, reaction component body 324 may include a rotary valve 330 and reagent reservoirs 332, which are fluidly coupled to rotary valve 330. Reaction component body 324 may also include additional reservoirs 334.
[087] The second subset 322 includes a light set 336, which includes several channels for directing light 338. Each channel for directing light 338 is optically coupled to a light source (not shown), such as a light emitting diode. led light). The light source (s) are configured to provide an excitation light, which is directed through the light directing channels 338 in the biosensors. In alternative embodiments, the cartridge may not include one or more light sources. In these embodiments, one or more light sources can be located on workstation 300. When the cartridge is inserted into the 306 system receptacle (Figure 4), cartridge 302 can align with one or more light sources , so that the biosensors can be illuminated.
[088] Also shown in Figure 6 is the second subset 322, which includes a cartridge pump 340, which is coupled by fluid communication with the orifices 342 and 344. When the first and second subsets 320 and 322 are joined together, orifice 342 is coupled to rotary valve 330 and orifice 344 is coupled to other reservoirs 334. Cartridge pump 340 can be activated to direct reaction components from reservoirs 332 and / or 334 to biosensors, according to an established protocol .
[089] Figure 7 illustrates a cross section of part of an exemplary biosensor 400, formed according to one embodiment. The biosensor 400 can include items similar to those in biosensor 102 (Figure 1), described above, and can be used, for example, in cartridge 302 (Figure 4). As shown, biosensor 400 can include a flow cell 402, which is coupled, directly or indirectly, to a detector device 404. The flow cell 402 can be mounted on the detector device 404. In the illustrated embodiment, the flow cell 402 it is attached directly to the detector device 404 by one or more securing mechanisms (for example, adhesives, connectors, fasteners and the like). In some embodiments, flow cell 402 can be removably coupled to detector device 404.
[090] In the illustrated embodiment, the detector device 404 includes a device base 425. In particular embodiments, the device base 425 includes several stacked layers (for example, a silicon layer, a di-electric layer, metallic dielectric layers, etc.). The device base 425 may include a set of sensors 424 of light sensors 440, a guide arrangement 426 of light guides 462, and a reaction arrangement 428 of reaction recesses 408, which have the corresponding reaction sites 414. In certain embodiments, the components are arranged so that each light sensor 440 aligns with a single light guide 462 and a single reaction site 414. However, in other embodiments, a single light sensor 440 can receive photons per more than one light guide 462 and / or more than one reaction site 414. As used in this specification, a single light sensor can include one pixel or more than one pixel.
[091] Furthermore, it should be noted that the term "set" or "subset" does not necessarily include all items of a certain type that the detector device may have. For example, sensor array 424 may not include all light sensors in detector device 404. Instead, detector array 404 may include other light sensors (for example, one or more other light sensor sets). As another example, the guide arrangement 426 may not include all of the light guides of the detector device. Instead, there may be other light guides, which are configured differently from the 462 light guides, or which have different relationships to other elements of the detector device 404. As such, unless explicitly stated otherwise, the term "set "may or may not include all of these items from the detector device.
[092] In the illustrated embodiment, flow cell 402 includes a side wall 406, and a flow cover 410, which is supported by side wall 406 and other side walls (not shown). The side walls are coupled to the detection surface 412 and extend between the flow cover 410 and the detection surface 412. In some embodiments, the side walls are formed from a curable adhesive layer, which connects the flow cover 410 to the device detector 404.
[093] Flow cell 402 is dimensioned and formed so that a flow channel 418 exists between the flow cover 410 and the detector device 404. As shown, the flow channel 418 can include a height Hi. Only by for example, the height Hi can be between about 50 - 400 gm (microns), or, more particularly, about 80 - 200 gm. In the illustrated embodiment, the height Hi is about 100 gm. The flow cover 410 may include a material, which is transparent to the excitation light 401, which propagates from an external part of the biosensor 400 to the flow channel 418. As shown in Figure 7, the excitation light 401 approaches the flow cover 410 at a non-orthogonal angle. However, this is for illustrative purposes only, as the excitation light 401 can approach the flow cover 410 from different angles.
[094] Flow cover 410 is also shown, which may include inlet and outlet holes 420, 422, which are configured to couple other holes in fluid communication (not shown). For example, the other holes can be for cartridge 302 (Figure 4) or workstation 300 (Figure 4). The flow channel 418 is sized and formed to direct a fluid along the detection surface 412. The height Hi and the other dimensions of the flow channel 418 can be configured to maintain a substantially uniform flow of a fluid across the surface of the flow. detection 412. The dimensions of the flow channel 418 can also be configured to control the formation of bubbles.
[095] The side walls 406 and the drain cover 410 can be separate components, which are coupled together. In other embodiments, the side walls 406 and the flow cover 410 can be integrally formed so that the side walls 406 and the flow cover 410 are formed of a continuous piece of material. For example, the flow cover 410 (or the flow cell 402) can comprise a transparent material, such as glass or plastic. The flow cover 410 can constitute a substantially rectangular block, having a flat outer surface and a flat inner surface, which defines the flow channel 418. The block can be mounted on the side walls 406. Alternatively, the flow cell 402 can be engraved to define the flow cover 410 and the side walls 406. For example, a recess can be engraved on the transparent material. When the recorded material is mounted on the detector device 404, the recess can become the flow channel 418.
[096] Detector device 404 has a detection surface 412, which can be functionalized (for example, chemically or physically modified in a manner suitable to conduct the desired reactions). For example, the detection surface 412 can be functionalized and can include several reaction sites 414, having one or more biomolecules immobilized in them. The detection surface 412 has a set of reaction recesses or open-sided reaction chambers 408. Each reaction recess 408 can be defined by, for example, a notch or a change in depth along the detection surface 412. In other embodiments, the detection surface 412 can be substantially flat.
[097] As shown in Figure 7, reaction sites 414 can be distributed in a model along the detection surface 412. For example, reaction sites 414 can be located in rows and columns along the detection surface 412 in a way that is similar to a microcontroller. However, it should be understood that several models of reaction sites can be used. Reaction sites can include biological or chemical substances that emit light signals. For example, chemical or biological substances at reaction sites can generate light emissions in response to excitation light 401. In particular embodiments, reaction sites 414 include clusters or colonies of biomolecules (eg, oligonucleotides), which are immobilized on the detection surface 412.
[098] Figure 8 is an enlarged cross section of the detector device 404, showing several items in more detail. More specifically, Figure 8 shows a single light sensor 440, a single light guide 462 for directing light emissions towards the light sensor 440, and an associated 446 circuitry for transmitting signals based on light emissions ( for example, photons), detected by the light sensor 440. It should be understood that other light sensors 440 from the sensor set 424 (Figure 4) and associated components can be configured in an identical or similar way. It should also be understood, however, that the detector device 404 does not need to be produced in a completely identical or uniform manner. Instead, one or more 440 light sensors and / or associated components can be produced differently or have different relationships with each other.
[099] Circuitry 446 may include conductive elements (for example, conductors, lines, routes, interconnections, etc.), which are capable of conducting electrical current, such as the transmission of data signals, which are based on photons detected. For example, in some embodiments, circuitry 446 may be similar or include an array of microcircuits, such as the array of microcircuits described in U.S. Patent 7,595,883, which is incorporated in this specification by reference in its entirety. The detector device 404 and / or the device base 425 can or can comprise an integrated circuit, having a flat arrangement of the light sensors 440. The circuitry 446, formed within the detector device 425, can be configured for at least one amplification, digitization, storage and signal processing. The circuitry can collect and analyze the detected light emissions and generate data signals, to communicate the detection data to a bioassay system. Circuitry 446 can also perform digital and / or analog signal processing on detector device 404.
[0100] The device base 425 can be produced by using integrated circuit production processes, such as the processes used for the production of complementary metal oxide semiconductors (CMOSs). For example, the device base 425 may include several stacked layers 431 - 437, including a layer or a sensor base 431, which is a layer or silicon wafer in the illustrated embodiment. The sensor layer 431 can include the light sensor 440 and ports 441 - 443, which are formed with the sensor layer 431. Ports 441 - 443 are electrically coupled to the light sensor 440. When the detector device 404 is formed entirely as shown in Figures 7 and 8, light sensor 440 can be electrically coupled to circuitry 446 through ports 441 - 443.
[0101] As used in this specification, the term "layer" is not limited to a single continuous body of material, unless otherwise indicated. For example, the sensor layer 431 can include multiple sublayers, which are of different materials and / or can include coatings, adhesives and the like. In addition, one or more of the layers (or sublayers) can be modified (for example, engraved, deposited with material, etc.), to provide the items described in this specification.
[0102] In some embodiments, each light sensor 440 has a detection area, which is less than about 50 pm2. In particular embodiments, the detection area is less than about 10 pm2. In more particular embodiments, the detection area is about 2 pm2. In such cases, the light sensor 440 may constitute a single pixel. An average reading noise of each pixel in a 440 light sensor can be, for example, less than about 150 electrons. In more particular embodiments, the reading noise can be less than about 5 electrons. The resolution of the light sensor set 440 can be greater than about 0.5 megapixel (Mpixel). In more specific embodiments, the resolution can be greater than about 5 Mpixels and, more particularly, greater than about 10 Mpixels.
[0103] The device layers also include several metallic dielectric layers 432 - 437, which are referred to below as substrate layers. In the illustrated embodiment, each substrate layer 432 - 437 includes metallic elements (for example, W - tungsten, Cu - copper or Al - aluminum) and dielectric material (for example, SiO2). Various metallic elements and a dielectric material can be used, such as those suitable for integrated circuit production. However, in other embodiments, one or more of the layers of sb, one or more of the layers of substrate 432 - 437 can include only dielectric material, such as one or more layers of SiO2.
[0104] With respect to the specific embodiment shown in Figure 8, the first layer of substrate 432 may include metallic elements, referred to as M1, that are embedded within the dielectric material (for example, SiO2). The metallic elements M1 comprise, for example, W (tungsten). The metallic elements M1 extend over the substrate layer 432 in the illustrated embodiment. The second layer of substrate 433 includes metallic elements M2 and dielectric material, as well as metallic interconnections (M2 / M3). The third layer of substrate 434 includes the metallic elements M3 and the metallic interconnections (M2 / M4). The fourth layer of substrate 435 also includes the metal elements M4. The device base 425 also includes a fifth and a sixth layer of substrate 436, 437, which are described in more detail below.
[0105] As shown, the metallic elements and the interconnections are connected together, to form at least part of the 446 circuitry. In the illustrated embodiment, the metallic elements M1, M2, M3, M4 include W (tungsten), Cu (copper) and / or Al (aluminum), and the metallic interconnections M2 / M3 and M3 / M4 include W (tungsten), but it should be understood that other materials and configurations can be used. It should also be noted that device base 425 and detector device 404, shown in Figures 7 and 8, are for illustrative purposes only. For example, other embodiments may include more or less layers than those shown in Figures 7 and 8 and / or different configurations of metallic elements.
[0106] In some embodiments, the detector device 404 includes a shield layer 450, which extends along an outer surface 464 of the device base 425. In the illustrated embodiment, the shield layer 450 is deposited directly across the surface outer layer 464 of substrate layer 437. However, an intermediate layer may be arranged between substrate layer 437 and shield layer 450, in other embodiments. The shield layer 450 can include a material, which is configured to block, reflect and / or significantly attenuate the light signals, which propagate from the flow channel 418. The light signals can be the excitation light 401 and / or light emissions 466 (shown in Figure 9). For example only, the shield layer 450 may comprise tungsten (W).
[0107] As shown in Figure 8, the shield layer 450 includes an opening or passage 452 through it. The shield layer 450 may include an arrangement of these openings 452. In some embodiments, the shield layer 450 may extend continuously between adjacent openings 452. As such, the light signals from flow channel 418 can be blocked, reflected and / or significantly attenuated, to prevent the detection of these light signals by light sensors 440. However, in other embodiments, the shield layer 450 does not extend continuously between adjacent openings 452, so that one or more openings different from those openings 452 exist in the shield layer 450.
[0108] The detector device 404 may also include a passivation layer 454, which extends over the shield layer 450 and through the openings 452. The shield layer 450 can extend through the openings 452, thereby covering, directly or indirectly, the openings 452. The shield layer 450 can be located between the passivation layer 454 and the device base 425. An adhesive or promoter layer 458 can be located between them to facilitate the coupling of the passive and shield layers. 454, 450. Passivation layer 454 can be configured to protect device base 425 and shield layer 450 from the fluid environment of flow channel 418.
[0109] In some cases, the passivation layer 454 can also be configured to provide a solid surface (i.e., the detection surface 412), which allows biomolecules or others of interest to be immobilized on it. For example, each reaction site 414 may include a cluster of biomolecules, which are immobilized on the detection surface 412 of the passivation layer 454. In this way, the passivation layer 454 can be formed of a material, which allows the reaction 414 are immobilized on it. The passivation layer 454 can also comprise a material, which is at least transparent to a desired fluorescent light. By way of example, the passivation layer 454 may include silicon nitride (SisN4) and / or silica (SiO2). However, another or other suitable materials can be used. In addition, the passivation layer 454 can be modified physically or chemically, to facilitate the immobilization of biomolecules and / or facilitate the detection of light emissions.
[0110] In the illustrated embodiment, a part of the passivation layer 454 extends along the shield layer 450, and a part of the passivation layer 454 extends directly along the filter material 460 of a light guide 462. The recess reaction 408 can be formed directly on the light guide 462. In some cases, before the passivation layer 454 is deposited along the shield layer 450 or the adhesion layer 458, a hole or base cavity 456 can be formed within the device base 452. For example, the device base 425 can be engraved to form an arrangement of the base holes 456. In particular embodiments, the base hole 456 is an elongated space, which extends close to the opening 452 towards the light sensor 440. The base hole can extend longitudinally along a central longitudinal axis 468. A three-dimensional shape of the base hole 456 can be substantially cylindrical or conical in some concretes. etizations, so that a cross section taken along a plane, which extends on the page of Figure 9, is substantially circular. The longitudinal axis 468 can extend through a geometric center of the cross section. However, other geometries can be used in alternative embodiments. For example, the cross section can be substantially square or octagonal.
[0111] The filter material 460 can be deposited inside the base hole 456, after the base hole 456 is formed. The filter material 460 can form (for example, after curing) a light guide 462. The light guide 462 is configured to filter excitation light 401 and allow light emissions 466 to propagate through it towards the sensor. corresponding 440 light. The light guide 462 can be, for example, an organic absorption filter. By way of specific example only, the excitation light can be about 532 nm, and the light emissions can be about 570 nm or more.
[0112] In some cases, the organic filter material may be incompatible with other materials in the biosensor. For example, the organic filter material may have a thermal expansion coefficient, which causes the filter material to expand significantly. Alternatively or in addition, the filter material may be unable to adhere sufficiently to certain layers, such as the shield layer (or other metallic layers). The expansion of the filter material can cause mechanical stress in the layers, which are adjacent to the filter material or structurally linked to the filter material. In some cases, the expansion can cause cracks or other undesirable aspects in the structure of the biosensor. As such, the embodiments presented in this specification can limit the degree to which the filter material expands and / or the degree to which the filter material is in contact with other layers. For example, filter materials from different light guides can be isolated from each other by the passivation layer. In these embodiments, the filter material may not be in contact with the metal layer (s). In addition, the passivation layer can resist expansion and / or allow some expansion, while reducing the generation of undesirable structural aspects (eg cracks).
[0113] The light guide 462 can be configured relative to the material surrounding the device base 425 (for example, the dielectric material), to form a light guide structure. For example, the 462 light guide can have a refractive index of about 2.0, so that light emissions are substantially reflected at an interface between the 462 light guide and the device base material 425 In certain embodiments, the light guide 462 is configured so that the optical density (OD) or the absorbance of the excitation light is at least about 4 OD. More specifically, the filter material can be selected and the light guide 462 can be sized to obtain at least 4 OD. In more particular embodiments, the light guide 462 can be configured to obtain at least about 5 OD or at least about 6 OD. Other items of the biosensor 400 can be configured to reduce electrical and optical interference.
[0114] Figure 9 illustrates an enlarged view of the detection surface 412 and parts of the detector device 404 (Figure 7), which are located close to the detection surface 412. More specifically, the passivation layer 454, the adhesion layer 458, the shield layer 450 and the light guide 462 are shown in Figure 9. Each layer can have an outer surface (top) or an inner surface (bottom), and can extend along an adjacent layer in an interface. In some embodiments, the detection surface 412 is configured to form the reaction recess 408, near the opening 452. The reaction recess 408 can be, for example, a notch, a hole, a well, a groove, or an open-sided chamber or channel. Alternatively, the detection surface 412 can be flat, without the recesses shown in Figures 7-9. As shown, opening 452 is defined by an opening or layer edge 504. The layer edge 504 faces radially inward in the direction of longitudinal axis 468.
[0115] The detection surface 412 can include a raised portion 502, and the reaction recess 408 can include a base surface 490. The base surface 490 can extend substantially parallel to the shield layer 450. The detection surface 412 it may also include a side surface 492, which extends substantially orthogonal to the base surface 490 and to the elevated part 502 of the detection surface 412. The side surface 492 can define a periphery of the reaction recess 408. Although the elevated part 502, the base surface 490 and side surface 492 are referred to as separate surfaces, it should be understood that the surfaces may be part of the detection surface 412. Furthermore, it should be understood that, due to production tolerances, the surfaces may not be easily distinguished. For example, in other embodiments, the base surface 490 and the side surface 492 can be substantially a single surface with a concave shape.
[0116] The base surface 490 may represent (or include a point representing) a deeper part of the passivation layer 454, along the detection surface 412 within the reaction recess 408. For example, the raised part 502 may extend along a surface plane Pi, and the base surface 490 may extend along a surface plane P2. As shown, the surface planes Pi and P2 are displaced relative to each other by a depth or distance D 1. The superficial plane P2 is closer to the light guide 462 or the light sensor 440 (Figure 7) than the superficial plane Pi. In the illustrated embodiment, the depth D 1 of the base surface 490 is substantially continuous, because the base surface 490 is substantially flat. In another embodiment, however, the depth D 1 may vary. For example, the base surface 490 may be concave in shape, with the depth increasing as the base surface 490 extends towards a center or intermediate part of it.
[0117] Reaction recess 408 may extend towards, or be located within, opening 452. For example, at least a part of the base surface 490 may reside within opening 452. Shield layer 450 may have a outer surface 506, which faces passivation layer 454, and inner surface 508, which faces device base 425. Outer surface 506 may extend along a surface plane P3, and inner surface 508 can extend along a superficial plane P4. The distance between the surface planes P3 and P4 can represent the thickness of the shield layer 450. As shown, the surface plane P3 can be located between the surface planes Pi, P2. As such, the base surface 490 extends into opening 452, as defined by layer edge 504. In other embodiments, however, the surface plane P2 can be located above the surface plane P3, so that the base surface 490 does not reside within aperture 452. Furthermore, in some embodiments, the surface plane P2 may be located below the surface plane P4, so that the base surface 490 is located below aperture 452.
[0118] The passivation layer 454 includes the detection surface 412, and an inner surface 510, which extends along the outer surface 506 of the shield layer 450 on an interface 512. In some embodiments, the adhesion layer 458 can extend along, and define, interface 512, between the shield layer 450 and the passivation layer 454.
[0119] In the illustrated embodiment, the passivation layer 454 extends directly along the light guide 462. More specifically, the inner surface 510 of the passivation layer 454 can directly engage with a material surface 514 of the light guide 462 As used in this specification, the term "coupling directly" and the like may include the two layers directly in contact with each other, or the two layers being bonded together using one or more adhesion promoting materials. The light guide 462 has an entrance region 472, which includes the material surface 514. The entrance region 472 can represent a part of the light guide 462, which initially receives light emissions.
[0120] The inner surface 510 can be directly coupled to the material surface 514 on an interface 516. The interface 516 can represent a material level of the filter material 460, which is deposited inside the guide cavity 456 (Figure 7). In the illustrated embodiment, interface 516 is substantially flat, so that interface 516 extends along an interface plane Ps. The interface plane Ps can extend substantially parallel to one or more of the surface planes Pi, P2, P3, P4. In other embodiments, however, interface 516 may be concave in shape, so that interface 516 is arched in the direction of light sensor 440 (Figure 8), or in an opposite direction away from light sensor 440.
[0121] The passivation layer 454 can fill a void generated when opening 452 is formed. Thus, in some embodiments, the passivation layer 454 can be located within or reside in aperture 452. In particular embodiments, interface 516 can be located at a depth D2 on the device base 425. In particular embodiments, the depth D2 can be configured so that the interface 516 is located below the opening 452, as shown in Figure 8. In these embodiments, the passivation layer 454 can isolate (for example, separate) the filter material 460 and the shield layer 450. These embodiments they can be suitable when the filter material 460 and the shield layer 450 are incompatible, so that cracks, or other undesirable aspects, can develop during manufacture of use of the biosensor 400 (Figure 7). In other embodiments, at least a part of the interface 516 can be located within the opening 452.
[0122] It is also shown in Figure 9 that the passivation layer 454 can form a joint or corner region 519. The joint region 519 can include the side surface 492 and extend around the longitudinal axis 468. The region of joint 519 may include a relatively thicker part of the passivation layer 454, which extends from the raised part 502 to the inner surface 510 at the material interface 516 (or between the surface plane Pi and the interfacial plane Ps). The dimensions of the joint region 519 can withstand the mechanical stresses caused by expansion of the filter material 460, during the manufacture of the biosensor 400 and / or during the thermal cycling, which can occur during the desired protocols (for example, SBS sequencing). As shown, the thickness between the surface plane Pi and the interfacial plane P5 is more than twice the thickness between the raised portion 502 of the detection surface 412 and the interface 512.
[0123] Reaction site 414 can include biological or chemical substances, which are generically represented as points 520 in Figure 9. Biological or chemical substances can be immobilized on detection surface 412, or, more specifically, on base surfaces and sides 490, 492. In particular embodiments, the reaction site 414 is located near opening 452, so that light emissions propagate through the passivation layer 454, through opening 452 and to the entrance region 472 of the light guide 462.
[0124] In some embodiments, reaction sites 414 or biological or chemical substances 520 in them can be standardized, so that reaction sites 414 or substances 520 have predetermined locations. For example, after passivation layer 454 is applied, reaction sites 414, or parts of them, can be standardized on passivation layer 454. In the illustrated embodiment, each opening 452 is associated with a single reaction site 414, so that the light emissions from the reaction site 414 are directed towards the corresponding light sensor 440. Biological or chemical substances 520, in a single reaction site 414, can be similar or identical (for example, an oligonucleotide colony that has a common sequence). However, in other embodiments, more than one reaction site 414 can correspond to one of the openings 452.
[0125] In particular embodiments, reaction sites 414 may include pads or metal regions, which are described in provisional patent application US 61 / 495,266, filed on June 9, 2011 and in provisional patent application US 61 / 552,712, filed on October 28, 2011. Both provisional patent applications US 61 / 495,266 (patent application 266) and 61 / 552,712 (patent application 712) are incorporated into this specification by reference in their entirety. In some embodiments, reaction sites 414 can be manufactured after flow cell 402 (Figure 7) is manufactured in detector device 404.
[0126] In the illustrated embodiment, reaction site 414 includes a colony of oligonucleotides 520, in which the oligonucleotides have an effectively common sequence. In these embodiments, each of the oligonucleotides can generate common light emissions, when excitation light 410 is absorbed by the fluorophores incorporated within the oligonucleotides. As shown, 466 light emissions can emit in all directions (for example, isotropically), so that, for example, a part of the light is directed to the 462 light guide, a part of the light is directed to reflect from the layer shield 450, and a part of the light is directed to the flow channel 418 or to the passivation layer 454. For the part which is directed to the light guide 462, the embodiments described in this specification can be configured to facilitate the detection of photons.
[0127] Also shown in Figure 9 is the device base 425, which may include the peripheral interference shields 522, 524, located within the device base 425. The interference shields 522, 524 can be positioned relative to the light 462 and configured so that the interference shields 522, 524 block or reflect the light signals, which propagate outside the light guide 462. The light signals may include the excitation light 401, which has been reflected or refracted and / or light emissions 466 generated on or near the detection surface 412. In some embodiments, interference shields 522, 524 can also directly block excitation light 401 from flow channel 418. As such, the shields interferences 522, 524 can reduce the detection of unwanted light signals. For example, interference shields 522, 524 can reduce optical interference between adjacent light sensors 440 and / or can improve the collection efficiency of the corresponding light sensor 440. The interference shields 522, 524 can be, for example, metallic elements, which are manufactured during the manufacture of the device base 425. In some embodiments, the processes used to manufacture the elements M1, M2, M3, M2 / M3 and M3 / M4 of circuit set 446 (Figure 8) can be the same or similar to the processes that manufacture interference shields 522, 524. For example, interference shields 522, 524 can be located within the dielectric material (for example, layers dielectric) of the device base 425, and comprise the same material that is used to manufacture circuitry 446 (for example, one or more of the materials used to manufacture the elements M1, M2, M3, M2 / M3 and M3 / M4 ). Although not shown, in some cases, the different stages of manufacturing CMOS may include the formation of metallic elements, which will transmit data signals, during the formation of the interference shields.
[0128] Although interference shields 522, 524 can be manufactured in a similar manner as circuitry 446, interference shields 522, 524 can be separated electrically from circuitry 446. In other words, for some embodiments, interference shields 522, 524 may not transmit data signals. In other embodiments, however, the interference shields 522, 524 can be traces or other metallic elements, which are configured to transmit data signals. As also shown in Figure 9, interference shields 522, 524 can have different cross-sectional dimensions (for example, width, height or thickness) and shapes, and can also be made of different materials.
[0129] In the illustrated embodiment, the interference shields 522, 524 are coupled together, to form a single larger interference shield. However, the interference shields 522, 524 can be spaced apart in other configurations. For example, the interference shields 522, 524 can be spaced apart along the longitudinal axis 468. In the illustrated embodiment, the interference shields 522, 524 surround, at least partially, the entrance region 472 and part of the passivation 454. The interference shield 522 directly couples the shield layer 450. In some embodiments, the interference shields 522, 524 may only partially surround the light guide 462. In other embodiments, the interference shields 522, 524 may constitute interference rings, which circumferentially surround the entire 462 light guide. These embodiments are described in more detail below with respect to Figures 10 and 11.
[0130] As shown, the guide cavity 456 is defined by one or more internal surfaces 526 of the device base 425. In particular embodiments, the internal surfaces 526 can be one or more surfaces of the dielectric material (for example, SiO2) of the substrate layers 432 - 437. The interference shields 522, 524 can come directly into contact with the light guide, so that part of the metal elements are exposed and directly coupled to the filter material 460 of the light guide 462. In other embodiments, however, the interference shields 522, 524 are not exposed to the light guide 462, and, differently, can be positioned immediately adjacent to the light guide 462, so that a part of the dielectric material is located between the shields interference 522, 524 and the light guide 462. For example, in the illustrated embodiment, the dielectric material 528, 530 is located between the light guide 462 and the interference shields 522, 52 4, respectively. Each dielectric material 528, 530 may include a portion of the inner surface 526. Dielectric material 528, 530 may separate the light guide 462 from the respective interference shields 522, 524 by an SD separation distance. For example only, the separation distance SD is at least about 100 nm. The SD separation distance can be less than 100 nm.
[0131] Figure 10 is a schematic cross section of a detector device 602, formed according to another embodiment. Detector device 602 can include items similar to those of detector device 404 (Figure 7), and can be used in biosensors, such as biosensor 400 (Figure 7) or biosensor 102 (Figure 1). Detector device 602 can also be manufactured using integrated circuit manufacturing technologies. Detector device 602 is described and illustrated to demonstrate other items that detector and biosensor devices can have. In some embodiments, only detector device 602 can constitute a biosensor. In other embodiments, the detector device 602 can be coupled to a flow cell to form a biosensor. For example, the detector device 602 can be coupled to the flow cell 402 and form a shield layer between the detector device 602 and the flow cell 402.
[0132] As shown, the detector device 602 includes a device base 604, a shield layer 640 and multiple sublayers 652, 654, which collectively form a passivation layer 650 of the detector device 602. The device base 604 includes a set of sensors 606 of light sensors 608 and an array of guides 610 of light guides 612. Light sensors 608 may be similar or identical to light sensors 440, and light guides 612 may be similar or identical to guides of light 462. For example, light guides 612 are configured to receive excitation light 614 and light emissions 616. As shown, light emissions 616 are illustrated as light being emitted from a single point. It should be understood that light emissions can be generated from multiple points along the passivation layer 650. Each light guide 612 extends to the device base 604 along a central longitudinal axis 618, from an entry region 620 of the light guide 612 towards a corresponding light sensor 608 of the sensor set 606.
[0133] Similar to light guides 462, light guides 612 may include a filter material, which is configured to filter excitation light 614 and allows light emissions 616 to propagate through them towards the 608 light sensors corresponding. The device base 604 includes a device circuitry (not shown), which is electrically coupled to the light sensors 608 and configured to transmit photon-based data signals, detected by the light sensors. Although not shown in Figures 10 and 11, the circuitry of the device base 604 can be located between the light guides 612, similar to the circuitry 446 (Figure 8) located between the light guides 462.
[0134] As shown, device base 604 includes peripheral interference shields 631 - 634, which are located within device base 604. More specifically, each light guide 612 is surrounded by multiple interference shields 631 - 634. The interference shields 631 - 634 for each light guide 612 can be spaced apart along the respective longitudinal axis 618, so that the spans 641 - 643 are formed between them. The sizes of the spans 641 - 643 can be substantially the same or they can be different. For example, spans 643 are slightly larger than spans 642.
[0135] In the illustrated embodiment, the interference shields 631 - 634 are configured to circumferentially surround the 612 light guides. As used in this specification, the term "circumferentially surround" is not intended to require that the 612 light guides have a circular cross section and / or the interference shields 631 - 634 have circular shapes. In contrast, an interference shield can circumferentially surround the light guide 612 if the interference shield encircles the corresponding longitudinal axis 618. The interference shield can completely surround the longitudinal axis 618, or only partially surround the longitudinal axis 618. For example, the interference shields 631 - 634 can be extended continuously around the corresponding light guide 612, or, in other cases, interference shields 631 - 634 can include multiple sub-elements, which are individually distributed around the light guide 612, to surround, at least partially, the corresponding light guide.
[0136] Similar to the shield layer 452, the shield layer 640 can form openings 642 through it. The openings 642 are substantially aligned with the light guides 612 and the corresponding light sensors 608, to allow the light signals to propagate to the corresponding input regions 620. The sublayer 654 can be deposited on the shield layer 640, so that the material of the sublayer 654 fills at least a part of the openings. In some embodiments, an additional sublayer 652 is deposited on sublayer 654, to form passivation layer 650. Just by way of example, any of sublayers 652, 654 can include Ta2Os by plasma vapor deposition (PVD) or SixNy by deposition plasma-optimized chemical vapor (PECVD). In another embodiment, an additional sublayer can be stacked on sublayers 652, 654. By way of a specific example, sublayer 654 can be PVD Ta2Os, sublayer 652 can be PECVD SixNy, and an additional layer, which is stacked on sublayer 652, can be PV2 Ta2θs.
[0137] Figure 11 is a flowchart illustrating a process 700 for manufacturing a biosensor, according to one embodiment. Process 700 is illustrated in Figures 12A and 12B. Process 700, for example, can employ structures or aspects of the various embodiments (for example, systems and / or processes) discussed in this specification. In various embodiments, some steps can be omitted or added, some steps can be combined, some steps can be performed simultaneously, some steps can be carried out concurrently, some steps can be divided into multiple steps, some steps can be carried out in a different order , or some steps or series of steps can be carried out again in an iterative mode.
[0138] Process 700 may include providing (at 702) a device base 800, having a set of 802 light sensor sensors. As shown, device base 800 has an outer or outer surface 801. The device base 800 can be manufactured using integrated circuit manufacturing technologies, such as CMOS manufacturing technologies. For example, the device base 800 may include several layers of substrate with different modified items (e.g., metallic elements) embedded in them. In some embodiments, the device base 800 can include the guide regions 804 and the regions of the circuitry 806. The guide regions 804 can correspond to the parts of the device base 800, which will include, after process 700, the light guides. The adjacent guide regions 804 can be separated by the regions of the circuitry 806, which include the circuitry of the device (not shown), which may be similar to the circuitry described in this specification. More specifically, the circuitry of the device can be electrically coupled to the 802 light sensors and configured to transmit photon-based data signals, detected by the 802 light sensors. In some embodiments, the guide regions 804 may include the shields of peripheral interference 808, which surround the substrate material in the guide regions 804.
[0139] Process 700 may also include the application (at 704) of a shield layer 810 to the outer surface 801 of the device base 800 and the formation (at 706) of openings 812 by the shield layer 810. As described above, the shield layer 810 may include a metallic material, which is configured to block light signals. The openings 812 can be formed by applying a mask (not shown) and removing material (for example, by chemical attack) from the shield layer 810, to form the openings 812.
[0140] At 708, the guide cavities 814 can be formed on the device base 800. More specifically, the substrate material within the guide regions 804 can be removed so that the guide cavities 814 extend close to the openings 812 towards the corresponding light sensors 802. As shown in Figure 12A, the inner surfaces 815 of the substrate material can define the guide cavities 814. The guide cavities 814 can be dimensioned and formed so that the inner surfaces 815 are close to the interference shields 808. As described in In this specification, interference shields 808 can be immediately adjacent to internal surfaces 815, or can be exposed in guide cavities 814.
[0141] Process 700 may also include the deposition (at 710) of filter material 820 within the guide cavities 814. The filter material 820 can be, for example, an organic filter material. In some embodiments, a portion of the filter material 820 may extend along the shield layer 810, after the deposition operation. For example, the amount of filter material 820 applied to the device base 800 may exceed the volume available within the guide cavities 814. As such, the filter material 820 may overflow from the guide cavities 814 and extend along the shield layer. 810.
[0142] In some embodiments, the deposition (in 710) of the filter material 820 may include compression (for example, using a component like a juicer) of the filter material 820 in the guide cavities 814. Figure 12A appears to indicate a uniform layer of the filter material 820 along the shield layer 810. In some embodiments, the layer of filter material 820 may not be uniform. For example, only parts of the shield layer 810 can have filter material 820 in them. In alternative embodiments, the deposition operation may include selective filling of each of the guide cavities 814, so that the filter material 820 does not empty or overflow from the guide cavities 814.
[0143] In 712, the filter material 820 can be cured. Optionally, process 700 may also include removing (at 714) filter material 820 from shield layer 810, and, in some cases, parts of filter material 820 from guide cavities 814. Filter material 820 can be removed from within of the guide cavities 814, so that a material level 830 of the filter material 820 is located within the opening 812, or at a depth below the shield layer 810. In embodiments where the material level 830 is below the layer of shield 810, filter material 820 may not come into contact with any material in shield layer 810. Filter material 820 within guide cavities 814 may form light guides. Different processes can be implemented for removing the filter material 820 from the shield layer 810. For example, the removal operation can include at least one chemical attack of the filter material or chemical polishing of the filter material.
[0144] As shown in Figure 12B, process 700 can also include the application (in 716) of a passivation layer 832 in the shield layer 810 and in the filter material 820 of the light guides, so that the passivation layer 832 extends directly across the light sensors 810 and through the openings 812. The passivation layer 832 can extend directly across the light guides at the corresponding material interfaces 834, such as material interfaces 516 (Figure 9). In the illustrated embodiment, the passivation layer 832 has a flat detection surface 836. In other embodiments, the detection surface 836 can form an array of reaction recesses, such as reaction recesses 408 (Figure 7). The reaction recesses may extend towards, or be located in, the corresponding openings 812.
[0145] In some embodiments, the passivation layer 832 includes multiple sublayers 841 - 843. In particular embodiments, at least one of the sublayers 841 - 843 includes tantalum. For example, sublayer 841 can include tantalum pentoxide (Ta2θs), sublayer 842 can include a low temperature film (for example, silicon nitride - SixNiy) and sublayer 843, which can have the detection surface 836, can include tantalum pentoxide (Ta2θs). However, sublayers 841 - 843 are only provided as examples, and other passive layers can include less sublayers, more sublayers, or sublayers with different materials. In some cases, only a single sublayer is used for the passivation layer.
[0146] Optionally, process 700 may include providing (in 718) reaction sites 850 and mounting a flow cell (not shown). The provision of reaction sites 850 can occur before or after the flow cell is coupled to the detector device. Reaction locations 850 can be located at desired addresses, so that reaction locations 850 have a predetermined pattern along the detection surface 836. Reaction locations can match (for example, a location for a light sensor, one location for multiple light sensors, or multiple locations for a light sensor) in a predetermined manner. In other embodiments, reaction sites may be formed randomly along the detection surface 836. As described in this specification, reaction sites 850 may include biological or chemical substances immobilized on the detection surface 836. Biological or chemical substances can be configured to emit light signals in response to the excitation light. In particular embodiments, reaction sites 850 include agglomerates or colonies of biomolecules (e.g., oligonucleotides), which are immobilized on the detection surface 836.
[0147] In one embodiment, a biosensor is provided, which includes a flow cell and a detector device having the flow cell coupled to it. The flow cell and the detector device form a flow channel, which is configured to have biological or chemical substances in it, which generate light emissions in response to an excitation light. The detector device includes a device base, having a set of light sensor sensors and an array of light guide guides. The light guides have input regions, which are configured to receive the excitation light and the runoff light emissions. The light guides extend on the device base of the input regions, in the direction of the corresponding light sensors, and have a filtering material, which is configured to filter the excitation light and allow light emissions to propagate in the direction of the corresponding light sensors. The device base includes the device circuitry electrically coupled to the light sensors, and is configured to transmit data signals based on photons detected by the light sensors. The detector device also includes a shield layer, which extends between the flow channel and the device base. The shield layer has openings, which are positioned relative to the corresponding light guide entry regions, so that light emissions propagate through the openings in the corresponding entry regions. The shield layer extends between the adjacent openings and is configured to block the excitation light and light emissions incident on the shield layer between the adjacent openings.
[0148] In one aspect, the entry regions of the light guides can be located within the corresponding openings of the shield layer, or they can be located at a depth in the device base.
[0149] In another aspect, the detector device may include a passivation layer, which extends along the shield layer, so that the shield layer is between the passivation layer and the device base. The passivation layer can extend through the openings.
[0150] In particular cases, the light guide filter material may be an organic filter material. The passivation layer can extend directly along the entrance regions of the light guides and isolate the organic filter material from the shielding layer. The material interfaces can be located within the corresponding openings in the shield layer, or located at a depth in the device base. In some embodiments, the passivation layer extends into the openings and forms an arrangement of reaction recesses. The reaction recesses can extend in the direction or be located inside the corresponding openings.
[0151] In some embodiments, biological or chemical substances are configured so that they are located within the reaction recesses. In some embodiments, the reaction recesses have corresponding base surfaces. The base surfaces can be located inside the opening or located at a depth in the device base.
[0152] In another aspect, the device base includes peripheral interference shields. Each interference shield can surround one of the corresponding light guides. The interference shields can be configured to reduce optical interference between the adjacent light sensors.
[0153] In another aspect, the biosensor is devoid of a lens, so the biosensor does not include an optical element that focuses light emissions towards a focal point.
[0154] In one embodiment, a biosensor is provided, which includes a flow cell and a detector device having the flow cell coupled to it. The flow cell and the detector device form a flow channel, which is configured to have biological or chemical substances in it, which generate light emissions in response to an excitation light. The detector device includes a device base, having a set of light sensor sensors and an array of light guide guides. The light guides are configured to receive the excitation light and the light emissions from the flow channel. Each light guide extends to the device base along a central longitudinal axis, from a light guide entry region towards a corresponding light sensor from the sensor set. The light guides include a filter material, which is configured to filter the excitation light and allow light emissions to propagate through it towards the corresponding light sensors. The device base includes a circuitry of the device, which is electrically coupled to the light sensors and configured to transmit data signals based on photons detected by the light sensors. The device base includes peripheral interference shields on it, which surround the corresponding light guides of the guide arrangement. The interference shields surround, at least partially, the corresponding light guides around the respective longitudinal axis, to reduce the optical interference between the adjacent light sensors.
[0155] In one aspect, the interference shields can surround the entry regions of the corresponding light guides.
[0156] In another aspect, the interference shields may include interference rings, which circumferentially surround the corresponding light guide.
[0157] In another aspect, the device base can include a complementary metal oxide semiconductor (CMOS), and the device circuitry. The interference shields include metallic elements, located within dielectric layers of the device base. The interference shields can be separated electrically from the device's circuitry.
[0158] In another aspect, the shield layer may extend between the flow channel and the device base. The flow channel may have openings, which are positioned relative to the light guide entry regions corresponding to the guide arrangement. The openings can allow light emissions to spread through them in the entrance regions. The shield layer can extend between the adjacent openings and is configured to block the excitation light and light emissions incident on the shield layer between the adjacent openings. For example, the entry regions of the light guides can be located within the corresponding openings of the shield layer or are located at a depth in the device base.
[0159] In another aspect, the detector device may also include a passivation layer, which extends along the shield layer, so that the shield layer is between the passivation layer and the device base and through the openings.
[0160] In another aspect, the interference shield comes into contact with, or is immediately adjacent to, the shield layer.
[0161] In another aspect, the interference shields are primary interference shields, and the device base includes secondary interference shields, where each light guide of the guide arrangement is at least partially surrounded by the primary interference shields corresponding secondary and secondary. For example, the primary and secondary interference shields can be spaced apart along the corresponding longitudinal axis. In another embodiment, the primary and secondary interference shields have different dimensions.
[0162] In one embodiment, a manufacturing process for a biosensor is provided. The process includes providing a device base, having a set of light sensor sensors and a set of device circuits that is electrically coupled to the light sensors, and which is configured to transmit data signals based on photons detected by the light sensors. light. The device base has an outer surface. The process also includes applying a shield layer to the outer surface of the device base and forming openings through the shield layer. The process also includes forming guide cavities, which extend from the corresponding openings towards a corresponding light sensor from the sensor set, and depositing filter material within the guide cavities. A part of the filtering material extends along the shield layer. The process also includes curing the filter material and removing the filter material from the shield layer. The filter material inside the guide cavities forms light guides. The process also includes applying a passivation layer to the shield layer, so that the passivation layer extends directly along the shield layer and through the openings.
[0163] In one aspect, removing the filtering material from the shield layer includes removing a portion of the filtering material from within the guide cavities, so that a level of filtering material material is located within the opening or at a depth below the shield layer.
[0164] In another aspect, the passivation layer extends directly along the light guides at the corresponding material interfaces. The material interfaces are located within the corresponding openings or located at a depth in the device base.
[0165] In another aspect, the filter material is an organic filter material. The passivation layer extends directly along the light guide and isolates the organic filter material from the shield layer.
[0166] In another aspect, the passivation layer forms an arrangement of reaction recesses. The reaction recesses extend towards, or are located within, the corresponding openings. For example, the reaction recesses can have corresponding base surfaces. The base surfaces can be located inside the opening or located at a depth in the device base.
[0167] In another aspect, the process includes coupling a flow cell to the device base, to form a flow channel between the passivation layer and the flow cell.
[0168] In another aspect, the removal of the filtering material from the shield layer includes at least one of chemical attack of the filtering material or of chemical polishing of the filtering material.
[0169] In another aspect, the passivation layer includes tantalum pentoxide (Ta2θδ). For example, the passivation layer can include multiple sublayers, where at least one of the sublayers includes tantalum pentoxide (Ta2θδ). In a more specific embodiment, the sublayers can include two layers of tantalum pentoxide with a low temperature film between them.
[0170] In another aspect, the device base has guide regions, which include substrate material, before the formation of the guide cavities, in which the adjacent guide regions are separated by regions of the circuit set, which include the set device circuits. The formation of the guide cavities may include removing the substrate material from the guide regions.
[0171] In another aspect, the device base may include peripheral interference shields, which surround the guide regions before the formation of the guide cavities. The interference shields can at least partially surround the corresponding light guides after the light guides are formed. The interference shields can be configured to reduce optical interference between the adjacent light sensors.
[0172] In one embodiment, a biosensor is provided, which includes a device base having a set of light sensor sensors and a guide arrangement of light guides. The device base has an outer surface. The light guides have input regions, which are configured to receive excitation light and light emissions generated by biological or chemical substances close to the outer surface. The light guides extend on the device base of the input regions, in the direction of the corresponding light sensors, and have a filtering material, which is configured to filter the excitation light and allow light emissions to propagate in the direction of the corresponding light sensors. The device base includes a circuitry of the device, which is electrically coupled to the light sensors and configured to transmit data signals based on photons detected by the light sensors. The biosensor also includes a shielding layer, which extends along the outer surface of the device base. The shield layer has openings, which are positioned relative to the entrance regions of the corresponding light guides, so that the light emissions propagate through the openings in the corresponding entrance regions. The shield layer extends between the adjacent openings and is configured to block the excitation light and light emissions incident on the shield layer between the adjacent openings.
[0173] In one embodiment, a biosensor is provided, which includes a device base having a set of light sensor sensors and a guide arrangement of light guides. The device base has an outer surface. The light guides are configured to receive excitation light and light emissions generated by biological or chemical substances close to the external surface. All light guides extend to the device base along a central longitudinal axis, from a light guide entry region towards a corresponding light sensor from the sensor set. The light guide includes a filter material, which is configured to filter the excitation light and allow light emissions to propagate through it towards the corresponding light sensors. The device base includes a circuitry of the device, which is electrically coupled to the light sensors and configured to transmit data signals based on photons detected by the light sensors. The device base includes peripheral interference shields on it, which surround the corresponding light guides of the guide arrangement. The interference shields, at least partially, surround the corresponding light guides around the respective longitudinal axis, to reduce the optical interference between the adjacent light sensors.
[0174] It should be understood that the object described in this specification is not limited in its application to the details of construction and the layout of the components presented in this specification, or illustrated in its drawings. The object described in this specification is capable of other embodiments, and of being practiced or conducted in various ways. Also, it should be understood that the phraseology and terminology used in this specification are for the purpose of description and should not be considered as limiting. The use of "including", "comprising" or "having", and their variations, is to mean that it covers the items listed below, and their equivalents, as well as additional items.
[0175] Unless otherwise indicated, the terms "assembled", "connected", "supported" and "coupled", and their variations, are used generically to cover both direct and indirect assemblies, connections, supports and couplings. Furthermore, "connected" and "coupled" are not limited to mechanical or physical connections or couplings. Also, it should be understood that the phraseology and terminology used in this specification with reference to the orientation of devices or elements (such as, for example, terms such as "above", "below", "front", "rear" , "distal", "proximal" and the like) are only used to simplify the description of one or more embodiments presented in it, not only indicate or imply that the referred device or element must have a particular orientation. In addition, terms such as "external" and "internal" are used in this specification for descriptive purposes and are not intended to indicate or imply relative importance or significance.
[0176] It should be understood that the description above is intended to be illustrative and not limiting. For example, the embodiments described above (and / or their aspects) can be used in combination with each other. In addition, many modifications can be made to adapt a particular situation or material to the teachings of the object described at the time, without departing from its scope. Although the dimensions, types of materials and coatings, described in this specification, are intended to define the parameters of the object described, they are by no means limiting and are exemplary embodiments. Many other embodiments will be evident to those skilled in the art by reviewing the description presented above. The scope of the inventive object must therefore be determined with reference to the appended claims, together with the full scope of equivalents, with which these claims are associated. In the appended claims, the terms "including" and "in which" are used as common English equivalents of the respective terms "comprising" and "in which". Furthermore, in the claims presented below, the terms "first", "second" and "third", etc. they are used merely as markers, and are not intended to impose numerical requirements on your objects. Furthermore, the limitations of the claims set out below are not written in a more intermediate function format, and should be interpreted based on Article 35 USC Ç 112, sixth paragraph, unless and until the limitations of the claims expressly use the term "means to", followed by an indication of the function void of another structure.
[0177] The claims presented below include aspects of certain embodiments of the inventive object and are considered to be part of the description presented above. These aspects can be combined with each other.

权利要求:
Claims (13)
[0001]
1. Biosensor, CHARACTERIZED by the fact that it comprises: a flow cell; and a detector device having the flow cell coupled to itself, the flow cell and the detector device forming a flow channel that is configured to have biological or chemical substances in it, which generate light emissions in response to an excitation light , the detector device including: a device base having a set of light sensor sensors and a set of light guide guides, the light guides having input regions that are configured to receive excitation light and emissions light from the flow channel, the light guides extending to the device base from the input regions towards the corresponding light sensors and having a filter material that is configured to filter the excitation light and allow the light emissions propagate towards the corresponding light sensors, the device base including a set of device circuits coupled to the light sensors and configured to transmit data signals based on photons detected by light sensors; and a shield layer extending between the flow channel and the device base, the shield layer having openings that are positioned in relation to the corresponding light guide inlet regions, so that light emissions propagate through the openings in the corresponding entry regions, the shield layer extending between adjacent openings and configured to block the excitation light and the light emissions incident on the shield layer between adjacent openings.
[0002]
2. Biosensor, according to claim 1, CHARACTERIZED by the fact that the entrance regions of the light guides are located inside the corresponding openings of the shield layer or are located at a depth in the device base.
[0003]
3. Biosensor, according to claim 1, CHARACTERIZED by the fact that it further comprises a passivation layer that extends along the shield layer so that the shield layer is between the passivation layer and the device base, the passivation layer extending through the openings.
[0004]
4. Biosensor, according to claim 3, CHARACTERIZED by the fact that the light guide filter material is an organic filter material, the passivation layer extending directly along the light guide entrance regions and isolating the material shielding layer organic filter.
[0005]
5. Biosensor, according to claim 3, CHARACTERIZED by the fact that the passivation layer extends directly along the entrance regions of the light guides in corresponding material interfaces, the material interfaces being located inside the corresponding openings of the shield layer or located at a depth in the device base.
[0006]
6. Biosensor, according to claim 3, CHARACTERIZED by the fact that the passivation layer extends into the openings and forms an arrangement of reaction recesses, the reaction recesses extending towards, or being located within, the corresponding openings .
[0007]
7. Biosensor, according to claim 6, CHARACTERIZED by the fact that the reaction recesses have corresponding base surfaces, the base surfaces being located inside the opening or located at a depth in the device base.
[0008]
8. Biosensor, according to claim 1, CHARACTERIZED by the fact that the device base includes peripheral interference shields, each interference shield surrounding a respective light guide among the corresponding light guides, the configured interference shields to reduce optical interference between adjacent light sensors.
[0009]
9. Biosensor, according to claim 3, CHARACTERIZED by the fact that the passivation layer includes tantalum pentoxide (Ta2Os).
[0010]
10. Biosensor, according to claim 9, CHARACTERIZED by the fact that the passivation layer includes multiple stacked sublayers, in which at least one of the sublayers includes tantalum pentoxide (Ta2Os).
[0011]
11. Biosensor, according to claim 10, CHARACTERIZED by the fact that the sublayers include two layers of tantalum pentoxide (Ta2Os) with a low temperature film between them.
[0012]
12. Biosensor, according to claim 10, CHARACTERIZED by the fact that the sublayers include two layers of tantalum pentoxide by plasma vapor deposition (PVD) with a silicon nitride film by chemical vapor deposition optimized with plasma ( PECVD) among them.
[0013]
13. Method for producing a biosensor, CHARACTERIZED by the fact that it comprises: providing a device base having a set of light sensor sensors and a set of device circuits that is electrically coupled to the light sensors and configured to transmit signals from photon-based data detected by light sensors, the device base having an outer surface; apply a layer of armor to the outer surface of the device base; forming openings through the shield layer; forming guide cavities extending from the corresponding openings towards a corresponding light sensor of the sensor assembly; depositing filter material within the guide cavities, in which a part of the filter material extends along the shield layer; cure the filter material; removing the filtering material from the shielding layer, the filtering material within the guide cavities forming light guides; and applying a passivation layer to the shield layer so that the passivation layer extends directly along the shield layer and through the openings.

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法律状态:
2020-03-24| B06U| Preliminary requirement: requests with searches performed by other patent offices: procedure suspended [chapter 6.21 patent gazette]|

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2020-08-11| B09A| Decision: intention to grant [chapter 9.1 patent gazette]|

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2020-11-24| B16A| Patent or certificate of addition of invention granted [chapter 16.1 patent gazette]|Free format text: PRAZO DE VALIDADE: 20 (VINTE) ANOS CONTADOS A PARTIR DE 09/12/2014, OBSERVADAS AS CONDICOES LEGAIS. |

优先权:

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申请号 | 申请日 | 专利标题

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US201361914275P| true| 2013-12-10|2013-12-10|

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US61/914,275|2013-12-10|

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