• 专利附录
  • 专利说明
  • 权利要求
  • 类似技术
  • 同族专利
  • 引用文献
  • 法律状态
  • 优先权
    • 专利摘要:
    manufacture of nanopores using high electric fields. a method is provided for making a nanopore in a membrane. the method includes: applying an electrical potential across the membrane, where the value of the electrical potential is selected to induce an electric field that causes a leakage current through the membrane; the flow of monitoring current through the membrane while the electrical potential is being applied; detecting an abrupt increase in the leakage current through the membrane; and removing the electrical potential across the membrane in response to detecting an abrupt increase in the leakage current.
    公开号:BR112014027829B1
    申请号:R112014027829-6
    申请日:2013-05-07
    公开日:2021-03-23
    发明作者:Wing Hei Kwok；Vincent Tabard-Cossa；Kyle Alexander Zarkel Briggs
    申请人:The University Of Ottawa；
    IPC主号:
    专利说明:

    [0001] [001] This application claims the benefit of Provisional Application No. US 61 / 643,651, filed on May 7, 2012 and Provisional Application No. US 61 / 781,081, filed on March 14, 2013. The complete description of each of the above orders is hereby incorporated by reference. FIELD
    [0002] [002] The present disclosure refers to a nanopore production technique using high electric fields. HISTORIC
    [0003] [003] Nanotechnology is based on our ability to manipulate matter and produce the structure of devices on a nanoscale. Current solid-state production methods that achieve reproducibility of dimensional control at the nanoscale are often complex and involve the use of expensive infrastructure, operated by highly qualified personnel. For example, the problem of producing a hole on a molecular scale, or nanopore, in a fine insulating solid-state membrane requires the use of focused high-energy particles, whether produced by a dedicated ion beam machine (ion beam sculpture) ) or perforation by transmission electron microscopy (MET). Although these advances in nanoproduction have put the production of nanoscale devices with subnanometric scale control within the reach of academic laboratories, they are unsuitable for the mass production of holes in a membrane, to create nanopores. This represents a major barrier to the commercialization of any solid state nanopore technologies for health science applications, including rapid DNA sequencing.
    [0004] [004] This section provides basic information related to the present description, which is not necessarily state of the art. ABSTRACT
    [0005] [005] This section presents a general summary of the disclosure, and is not a comprehensive disclosure of its entire scope or all of its characteristics.
    [0006] [006] A method of producing a nanopore in a dielectric membrane immersed in a solution containing ions is provided. The method includes: applying an electrical potential across the membrane, where the value of the electrical potential is selected to induce an electric field, which causes a leakage current through the different insulation membrane; monitor the flow of current through the membrane while the electrical potential is being applied; detecting a sudden irreversible increase in the leakage current through the membrane; and removing the electrical potential across the membrane in response to the detection of a sudden increase in the leakage current, to stop the production of the nanopores.
    [0007] [007] In one aspect, an abrupt increase in leakage current is detected by comparing the value of the monitored current to a threshold and then the electrical potential is no longer applied when the value of the monitored current exceeds the threshold.
    [0008] [008] In another aspect, the membrane is arranged between two reservoirs filled with a fluid and, thus, it prevents the fluid from passing between the two reservoirs.
    [0009] [009] Equipment is also provided for the production of a nanopore in the membrane. The apparatus includes: two reservoirs fluidly coupled through a passage between them; a pair of electrodes electrically connected to a voltage source, such that an electrode is disposed in each of the two reservoirs and the pair of electrodes generates an electrical potential across the membrane; a current sensor electrically coupled to one of the electrodes and which can work to measure the current that passes between the two reservoirs; and a controller in interface with the current sensor, where the controller detects an abrupt increase in the measured current and, in response to the detection of an abrupt increase in the measured current, removes the voltage between the electrodes.
    [0010] [0010] Other fields of application will be evident from the description provided here. The description and specific examples in this summary are for illustrative purposes only and are not intended to limit the scope of this disclosure. DRAWINGS
    [0011] [0011] Figure 1 is a flow chart describing a method of producing a single nanopore according to the present invention;
    [0012] [0012] Figure 2A is a diagram that describes an example of a configuration for the production of a nanopore;
    [0013] [0013] Figure 2B is a diagram of an example fluid cell;
    [0014] [0014] Figure 3 is a schematic representation of an example of a current amplifier circuit that can be used to produce a nanopore;
    [0015] [0015] Figures 4A to 4D are diagrams that illustrate the production of a nanopore in a thin membrane by high electric fields;
    [0016] [0016] Figure 5 is a graph showing the leakage current in the cell in relation to the applied electric field;
    [0017] [0017] Figures 6 and 7 are graphs that illustrate a pore creation event in a membrane that has a thickness of 10 nm and 30 nm, respectively;
    [0018] [0018] Figure 8 is a graph that represents current-voltage curves for three independent nanopores produced in different membranes;
    [0019] [0019] Figures 9A, 9B and 9C are graphs that illustrate the time-pore creation as a function of the transmembrane potential for membranes with a thickness of 30 nm, the time-pore creation as a pH function for membranes with a thickness of 30 nm, and time-pore creation as a function of transmembrane potential for membranes with a thickness of 10 nm;
    [0020] [0020] Figure 10 is a scatter plot of the average current block normalized in relation to the total translocation time of multiple single molecule events;
    [0021] [0021] Figure 11 is a diagram that illustrates a technique for locating the formation of a nanopore; and
    [0022] [0022] Figure 12 is a diagram that illustrates another technique for locating the formation of a nanopore.
    [0023] [0023] The drawings described here are for illustrative purposes only of selected embodiments and not of all possible implementations, and are not intended to limit the scope of this disclosure. Corresponding reference numbers indicate corresponding parts throughout the various views of the drawings. DETAILED DESCRIPTION
    [0024] [0024] Figure 1 shows a simple and low cost method for the production of a single nanopore with subnanometric resolution (for example, from 1 to 100 nanometers) on a thin membrane. The method is based on applying a voltage across the thin membrane at 12 to generate an electric field high enough to induce a leakage current through the membrane. In some embodiments, the membrane is arranged between two reservoirs filled with fluid, such that the membrane separates the two reservoirs and prevents the flow of liquid between the two reservoirs. The current flow through the membrane is monitored at 14, while the electric field is being applied. The creation of a single nanopore (ie the fluid channel that covers the membrane) is indicated by an abrupt, irreversible increase in the leakage current. To detect the creation of the nanopore, the monitored current can be compared at 16 to a predetermined threshold. When the monitored current exceeds the threshold, the applied voltage is terminated at 18. Thus, the initial size of the nanopore can be defined mainly based on the value of the current threshold, although other factors that influence the size of the nanopore are additionally described below . Although reference is made to the formation of a nanopore, the techniques described here are more generally applicable to orifices of varying sizes.
    [0025] [0025] The schematic of an example configuration for the production of a nanopore is shown in Figures 2A and 2B. The configuration is generally composed of a fluidic cell 22; a pair of electrodes 24 electrically coupled to a current amplifier circuit 25; and a controller 26 in interface with the current amplifier circuit 25. The fluid cell 22 is further defined by two reservoirs 33 fluidly coupled through a passage 34 to each other, as can be seen better in figure 2B. The current amplifier circuit 25 works to create a potential between the electrodes and measure the current flow between the two reservoirs 33. In some embodiments, the controller 26 can be implemented by a data acquisition circuit 28 coupled to a computer personal computer 27 or other type of computing device. Thus, the configuration is similar to that which is commonly used for biomolecular detection in the nanopore detection field. The fluidic cell 22 and / or the entire system can be arranged in a Faraday cage 23 connected to earth to isolate electrical noise. Other production configurations of a nanopore are contemplated by this disclosure.
    [0026] [0026] In the configuration example, a silicon chip 31 is used to house a membrane 30. The silicon chip 31 is placed between two silicone joints 32 and then positioned between the two reservoirs 33 of the fluid cell 22. In some embodiments, fluid cell 22 is composed of polytetrafluoroethylene (PTFE), although other materials are contemplated. A small pressure is applied to the two joints 32 by the fluid cell 22, to tightly seal the contact. The two reservoirs 33 are filled with a fluid that contains ions, and the two electrodes 24 are inserted into the respective reservoirs of the fluid cells 22. Example configurations may include, but are not limited to, chloride-based saline solutions with Ag electrodes / AgCl or copper sulfate solutions with copper electrodes. The fluid can also be a non-aqueous solvent, such as 1M LiCl in ethanol. The fluid can be the same in both reservoirs and does not need to have an active corrosion action against the membrane material. Other types of fluids and the means of positioning the membrane between the two reservoirs are also considered, such as micro and nanofluid encapsulation.
    [0027] [0027] In some embodiments, one of the electrodes can be placed in direct contact with the surface of the membrane. In other embodiments, one of the electrodes can be a nanoelectrode, which is positioned to locate the electric field in the membrane and, thus, to locate the formation of pores in the membrane. It is also understood that more than two electrodes can be used. For example, an electrode can be placed in each reservoir with a third electrode placed in direct contact with the membrane surface. Other electrode arrangements are also contemplated by this disclosure.
    [0028] [0028] In some implementations, membrane 30 consists of a dielectric material such as silicon nitride (SiNx). The membranes are preferably thin, with a thickness of 10 nm or 30 nm, although membranes with different thickness are contemplated by this disclosure. Membranes made up of other dielectric materials, such as other oxides and nitrides, which are commonly used as dielectric materials for transistors, also fall within the scope of this disclosure. Likewise, atomically thin membranes can be made up of other materials such as graphene, boron nitride and the like. It is also contemplated that the membranes can be made up of several layers of materials, including dielectric materials and / or conductive materials.
    [0029] [0029] In an exemplary embodiment, the current amplifier circuit 25 is based on a simple operation amplifier circuit for reading and controlling voltage and current, as shown in Figure 3. Operational amplifiers are powered by a voltage source of ± 20 volts. In operation, the circuit admits a command voltage (Vcommand) between ± 10 volts from a computer-controlled data acquisition card, which is amplified at ± 20 volts, and defines the potential across the membrane. The applied potential (Vout) can also be measured by the current amplifier circuit 25. The current flow between the two electrodes is measured on one or both electrodes with a sensitivity of pA. More specifically, the current is measured with a transimpedance amplifier topology. The measured current signal (Iout) is digitized by the data acquisition circuit and fed continuously to the controller. In this way, the current is monitored in real time by the controller, for example at a frequency of 10 Hz, although a faster sampling rate can be used for the faster response time, once the current reaches the threshold. Other circuit arrangements for applying a potential and measuring a current fall within the scope of this disclosure.
    [0030] [0030] In the exemplary embodiment, the current threshold is adjusted to coincide with the sudden increase in current, in order to define the minimum size of the nanopore in the order of 1 nm. In other embodiments, the nanopore size can be defined as being larger, however, while continuing to apply a potential across the membrane. That is, the size of the nanopore continues to increase as the monitored current continues to increase. Instead of setting the current threshold to coincide with the sudden increase in the leakage current, the current threshold value can be adjusted to different values to achieve a nanopore of varying sizes. An exemplary technique for fine-tuning the size of a nanopore is further described by Beamish et al. in "Precise Control of the Size and Noise of Solid-state Nanopores using High Electric Fields” (Precise control of the size and noise of solid-state nanopores using high electric fields) Nanotechnology 23 405301 (2012), which is incorporated here in its entirety by reference. Other techniques and arrangements for adjusting nanopores are also contemplated by this disclosure.
    [0031] [0031] In some embodiments, the electrical potential is removed from the membrane before the leakage current rises sharply (ie, before the formation of pores). For example, the electrical potential is removed after the monitored current exceeds a predefined threshold or after a certain period of time, but before the sudden increase in the leakage current. In this way, the pore can be partially perforated or formed in the membrane. The same or different processes can then be used at a later time to complete the formation of pores.
    [0032] [0032] Figures 4A to 4D further illustrate the technique of producing a nanopore in membrane 41. For purposes of illustration, membrane 41 is composed of silicon nitride which has, for example, about 10 nm or 30 nm of thickness, t. More specifically, low stress SiNx (<250 MPa) can be deposited on 200 μm thick silicon substrate 42 by deposition of chemical vapor at low pressure (LPCVD). A 50 μm x 50 μm window on the back of a substrate is opened by an anisotropic chemical attack by KOH. The absence of pre-existing structural damage (for example, holes, nanofissure, etc.) is inferred from the fact that no current (ie, <10 pA) is measured through a membrane in low electric fields (for example, < 0.1 V / nm) before the nanopore production.
    [0033] [0033] A single nanopore can be produced by applying a constant potential, AV, through a 41 membrane. The value of the electrical potential (for example, four volts) is selected to induce an electric field, which causes a current of escape through the membrane. In this example, the electric potential produces an electric field, E, of the order of 0.5 V / nm, in the membrane, where the electric field is defined as E = ΔV / t, and approximates the breaking strength of the dielectric material . For other situations, it is predicted that the electric field will be greater than 0.1 V / nm and probably within the range of 0.2 to 1 V / nm. In these high field forces, there is a sustainable leakage current, Ifuga, through the membrane, which remains otherwise insulating in low fields. Ifuga increases rapidly with the intensity of the electric field, but it is usually in the range of dozens of nanoamperes for the operating conditions, as shown in Figure 5. It is understood that the magnitude of the electrical potential necessary for the creation of pores can vary depending on the material of the membrane, the thickness of the membrane, as well as other factors.
    [0034] [0034] With reference to Figure 4B, the dominant conduction mechanism in the dielectric membrane is attributed to a form of encapsulation assisted by electron trap provided by ions in solution, since the membrane is too thick for significant direct encapsulation of quantum mechanics and migration of impurities that cannot produce lasting currents. Direct migration of electrolyte ions is also unlikely, as for a given intensity of the electric field, higher leakage currents are observed for the thicker membranes. Free charges (electrons or holes) can be produced by redox reactions on the surface or by the ionization field of embedded ions. The number of loaded traps available (structural defects) defines the magnitude of the observed leakage current. The accumulation of charge traps produced by an electric field induced by broken connections or energetic encapsulation charge carriers leads to a highly localized conductive path and a discrete dielectric failure event, as shown in Figure 4C.
    [0035] [0035] In Figure 4D, a nanopore is formed by removing material from the membrane at the vanishing point. The creation of a single nanopore (ie, fluidic channel that covers the membrane) is indicated by a sudden irreversible increase in the leakage current, which is attributed to the appearance of ionic current. A feedback control mechanism is used to quickly terminate the applied potential when the current exceeds a predetermined limit, Cut. In an example of an embodiment, the threshold may be a fixed value (for example, 110 nA). In some embodiments, the threshold may vary, for example depending on the magnitude of the leakage current. For example, the threshold can be defined as a multiplier (for example, 1.5) of the leakage current before a pore is formed. In other embodiments, the threshold can be defined as a rate at which the monitored current is varying (for example, ~ 10 nA / s, for SiNx). It is readily understood that the threshold value can vary depending on the membrane material, the thickness of the membrane, as well as other factors.
    [0036] [0036] Figures 6 and 7 show such an event of creating a pore in a SiNx membrane with a thickness of 10 nm and 30 nm, respectively. These results indicate that the Icorte value helps to limit the initial size of the nanopore. A tight threshold can thus produce nanopores on the order of 2 nm in diameter or smaller. After the nanopore production event, the nanopore size can be enlarged with subnanometric precision through the application of moderate square pulses of alternating current electric field in the range of ± 0.2 to 0.3 V / nm. This allows the nanopore size to be precisely adjusted for a particular detection application.
    [0037] [0037] To infer the nanopore size after production, its ionic conductance, G, can be measured and related to an effective diameter, d, assuming a cylindrical geometry and responsible for access resistance using
    [0038] [0038] I-V curves are performed in a window of ± 1 V, where the leakage current can be safely ignored. Figure 8 reveals an ohmic electrical response. Most nanopores have linear I-V curves and low 1 / f noise under production in high saline (for example, 1M KCI). The remaining nanopores that show signs of greater noise or self-barrier can be conditioned through the application of moderate electric field pulses, to amplify them a little, until the ohmic behavior is reached in high saline and neutral pHs. The I-V characteristics shown in Figure 8 imply a relatively symmetrical profile of the internal electrical potential of the pore, which supports the symmetric geometry with a uniform charge distribution of the surface assumed by the pore conductance model, also confirmed by MET images.
    [0039] [0039] The method described above can also be extended to produce rectifying nanopores with varying degrees of rectification (that is, behaving like diodes that pass current in one polarity, but not in another). For example, small nanopores (<3 nm) produced in highly acidic solutions (for example, 1M KCl with pH 2) can rectify. If the pore is noisy, the application of moderate square pulses of AC electric field (for example, low frequency pulses in the range of 0.2 to 0.3 V / nm for low stress silicon nitride) to condition the pore as well it must be made in an acidic solution. This preserves the grinding properties, even when the pore is enlarged. The degree to which a pore grinds can be increased by reducing the conductivity of the solution while maintaining the acidic pH, since the lower conductivity reduces sorting and makes surface effects more important. Other techniques for obtaining the rectification are also contemplated.
    [0040] [0040] On the other hand, the increase in conductivity or the advance towards neutral pH reduces the effects of rectification. In addition, the pore will lead in the opposite polarization to the one in which it was created, as long as the solution in the pore shares the same pH as the one in which it was created. Inverting the solution from acid to alkaline or vice versa, has been shown to invert the direction of rectification. Taking these observations into account, the pore geometry and surface load characteristics can be controlled by adjusting the pH of the solutions and the polarity of the tension across the membrane.
    [0041] [0041] To better characterize the nanopores, the noise in the ionic current is examined through measurements of power spectral density. Notably, the production method is capable of consistently producing nanopores with low 1 / f noise levels, comparable to completely wet nanopores drilled by MET. This can be attributed to the fact that nanopores are created directly in liquid instead of vacuum, and are therefore never exposed to air. So far, hundreds of individual nanopores have been produced (for example, ranging from 1 to 100 nm in size), with comparable electrical characteristics that are stable for several days. The pore stability can be maintained by storing the pores in high salt concentration (for example, LiCl at> 3M in water or LiCl at> 1M in ethanol).
    [0042] [0042] It is intriguing that the procedure described above triggers the production of a single nanopore, in particular in an aqueous KCl solution, which is not known to chemically etch SiNx at neutral pH, and given that the anodic oxidation of semiconductors or metals it is known for the production of nanopore matrices. In order to elucidate the mechanisms that lead to the formation of a pore in a dielectric membrane, the production process is further investigated as a function of applied voltage, membrane thickness, electrolyte composition, concentration and pH.
    [0043] [0043] Figures 9A and 9B show the time-pore creation as a function of the transmembrane potential for 30 nm thick membranes, in 1M KCl buffered at various pHs. Interestingly, the production time for a single nanopore increases exponentially with the applied voltage, and can be as short as a few seconds. For example, by increasing the voltage from 11 volts to 17 volts through a 30 nm thick SiNx membrane, the production time can be reduced 100 times. By increasing the voltage from four volts to 10 volts over a 10 nm thick membrane, the production time can be reduced up to 1,000 times. Thus, the increase in the applied electrical potential reduces the production time. The same is true for the applied electric field.
    [0044] [0044] The electrolyte composition also has a drastic effect on the production time. In 1M KCl in water, the production time for a 30 nm thick SiNx membrane decreased 10 times when going from pH 7 to pH 13.5 and decreased 1,000 times when going from pH 7 to pH 2, for a certain applied voltage. In a 10 nm thick SiNx membrane, the pH effect is less pronounced, with a maximum of 10-fold variation. Asymmetric pH conditions between the two sides of the membrane also greatly affect the production time, according to the voltage polarity (ie very fast production times are obtained for: cathode / negative side at high pH and anode / positive side at low pH, and very slow production times are obtained for cathode / negative side at low pH and anode / positive side at high pH).
    [0045] [0045] Figure 9C shows the time-pore creation as a function of the transmembrane potential for 10 nm thick SiNx membranes, buffered at pH 10, in various 1M Cl-based aqueous solutions. Production time again relates exponentially to the transmembrane potential, but the potential required for the formation of a nanopore will now be reduced by ~ 1/3, compared to membranes 30 nm thick, regardless of the different cations (K +, Na +, Li +) tested. The concentration levels of the electrolyte composition also had an effect on the production time. For example, on a 30 nm thick SiNx membrane, the production time at low concentrations (~ 10 mM KCl in water) was significantly increased (ie> 100 times) compared to high salt concentrations (KCl at ~ 10 mM) 1M and 3M in water).
    [0046] [0046] However, these observations indicate that the applied electric field, E = ΔV / t, through the membrane is the main driving force to start the production of a single nanopore. Fields in the range of 0.4 to 1 V / nm are close to the dielectric breaking strength of low stress SiNx films, and are fundamental to intensify the leakage current that is ultimately thought to cause collapse in the layers thin insulators. The exponential dependence on potential time-pore creation implies the same field dependency, which is reminiscent of the time-dielectric failure in door dielectrics. Thus, the mechanisms of dielectric rupture follow: (i) accumulation of charge traps (that is, structural defects) by rupture of the connection induced by the electric field or generated by the injection of charge from the anode or cathode, (ii) increase until a critical density forming a highly localized conductive path and (iii) causing physical damage due to substantial energy dissipation and the resulting heating. The process by which a single nanopore is produced in solution is similar, although damage to the nanoscale is controlled by limiting the localized leakage current at the beginning of the first discrete rupture event. Ultimately, a single nanopore is created since, for a fixed transmembrane potential, the nanopore formation path experiences an increase in the intensity of the electric field during growth, which locally reinforces the rate of defect generation. The process by which the material is removed from the membrane remains unclear, but broken bonds can be chemically dissolved by the electrolyte, or after conversion to oxides or hydrides. The dependence of pH on production time can be explained by the fact that the rupture at a low pH is amplified by impact ionization, producing an avalanche, due to the increased probability of injections of orifices or incorporation of H + from the anode, which increases the rate of structural defect formation.
    [0047] [0047] DNA translocation experiments are carried out to demonstrate that these nanopores can be used to benefit the detection of a single molecule. The electrophoretically driven passage of a DNA molecule through a membrane is designed to temporarily block the flow of ions in a way that reflects the length, size, charge and shape of the molecule. The results using a 5 nm diameter pore on a 10 nm thick SiNx membrane are shown in Figure 10. The scatter plot shows the event duration and the average current block of more than 2,400 single molecule translocation events of 5kb dsDNA. The current drop is indicative of the diameter of the molecule (~ 2.2 nm), while the duration represents the time it takes for a molecule to translocate completely through the pore. The characteristic shape of the events is indistinguishable from data obtained in nanopores perforated by MET. The observed quantized current blockages strongly support the presence of a single nanopore that covers the membrane. Using dsDNA (~ 2.2 nm in diameter) as a molecular size ruler, the value of single level blocking events ΔG = 7.4 ± 0.4nS, provides an effective pore diameter, consistent with the size extracted at from the pore conductance model.
    [0048] [0048] The production of nanopores by controlled dielectric breakage in solution represents a major step forward compared to current production methods, and could provide a path to commercialization of nanopore technology, allowing for mass production of devices at low cost . While the nanopore creation process is suspected to be an intrinsic property of the dielectric membrane, so that the nanopore can form anywhere on the membrane surface, current understanding strongly suggests that the position of the pore can be controlled by control location of the electric field strength or material dielectric strength. For example, this can be achieved by local thinning of the membrane, as indicated in 110, in Figure 11. The electric field across the membrane can be calculated as E = V / L, where V is the applied voltage and L is the thickness of the membrane. The voltage threshold of the dielectric failure is therefore three times lower for the diluted region - ensuring that the nanopore is more likely to be created in this region, since the production time is exponentially related to the field strength.
    [0049] [0049] In another example, the electric field can be confined to specific zones on the membrane, for example, using encapsulation by nano or microfluidic channel. Referring to Figure 12, nano or microfluidic channels 120 are defined on the upper side of membrane 41. Each channel is independently addressable, both fluidically and electrically. Four electrodes would be used on the upper side; while only one electrode is needed on the bottom side. A nanopore can be produced in each channel independently and as needed, because the electric field is confined to the regions within the channel. This approach also allows for the simple integration of independently addressable nanopores in a matrix format on a single chip. It is envisaged that other techniques can be employed to concentrate the electric field in specific areas, such as placing electrodes in direct contact with the surface of the membrane.
    权利要求:
    Claims (24)
    [0001]
    Method for making a single nanopore on a membrane (30, 41) and that includes providing the membrane on which the nanopore is manufactured, comprised of a dielectric material; characterized by the fact of understanding in addition: select an electric potential that induces an electric field in the membrane, where the electric field has a value greater than 0.1 volts per nanometer; apply the electrical potential across the membrane (12); monitor the leakage current through the membrane while the electrical potential is being applied through the membrane (14); detecting an abrupt increase in the leakage current through the membrane while the electrical potential is being applied through the membrane (16); and removing the electrical potential across the membrane in response to detecting a sudden increase in the leakage current to stop pore manufacturing (18).
    [0002]
    Method according to claim 1, characterized in that it also comprises selecting an electrical potential that approximates the dielectric strength of the membrane material.
    [0003]
    Method according to claim 1, characterized in that it detects an abrupt increase in the leakage current, also comprising determining a rate of change of the monitored current and comparing the rate of change to a limit.
    [0004]
    Method, according to claim 3, characterized in that it also includes removing the electrical potential when the rate of change of the monitored current exceeds the limit, thereby stopping manufacturing.
    [0005]
    Method according to claim 1, characterized by detecting an abrupt increase in the leakage current comprising also comparing a value of the monitoring current to a limit and removing the electrical potential when the value of the monitoring current exceeds a limit, thus stopping manufacturing.
    [0006]
    Method according to claim 1, characterized in that it also comprises arranging the membrane between the two reservoirs filled with a fluid containing ions, so that the membrane separates the two reservoirs and prevents the fluid from passing between the two reservoirs; place an electrode (24) in each of the two reservoirs; and generate the electrical potential using the electrodes.
    [0007]
    Method according to claim 6, characterized in that it also comprises placing one of the two electrodes in direct contact with the membrane.
    [0008]
    Method according to claim 1, characterized in that it also comprises increasing the electrical potential applied to reduce the manufacturing time.
    [0009]
    Method according to claim 1, characterized in that it also comprises increasing the electric field in the membrane to reduce the manufacturing time.
    [0010]
    Method according to claim 6, characterized in that it also comprises increasing the concentration of ions in the fluid to reduce the time of manufacture.
    [0011]
    Method according to claim 6, characterized in that it also comprises increasing the acidity of the fluid to reduce the time of manufacture.
    [0012]
    Method according to claim 6, characterized in that it also comprises increasing the alkalinity of the fluid to reduce the manufacturing time.
    [0013]
    Method according to claim 6, characterized in that it also comprises changing the acidity of the fluid in one reservoir with respect to the fluid in the other reservoir to change the manufacturing time.
    [0014]
    Method according to claim 6, characterized in that it also comprises adjusting the acidity of the fluid on each side of the membrane to control the geometry and surface charge characteristic of the pore.
    [0015]
    Method according to claim 6, characterized in that it also comprises adjusting the polarity of the electrical potential in the membrane to control the geometry and surface charge characteristic of the pore.
    [0016]
    Method according to claim 1, characterized in that it also comprises selecting an electric potential that induces an electric field having a value in the range of 0.1 to 1.0 volts per nanometer.
    [0017]
    Method according to claim 1, characterized in that the fluid is defined as ions dissolved in an aqueous solvent.
    [0018]
    Method according to claim 1, characterized in that the fluid is defined as ions dissolved in an organic solvent.
    [0019]
    Method according to claim 1, characterized in that the fluid does not need to have a specific chemical notch action towards the membrane.
    [0020]
    Method according to claim 1, characterized in that it also comprises controlling the location of the pore formed in the membrane by locating the resistance of the electric field or dielectric resistance near a desired location for the pore.
    [0021]
    Method according to claim 1, characterized in that it also comprises controlling the location of the pore formed in the membrane by reducing the thickness of the membrane in the area of a desired location for the pore.
    [0022]
    Method according to claim 1, characterized in that it also comprises controlling the rectification of the pore formed in the membrane based on the acidity and conductivity of the fluid.
    [0023]
    Mechanism to manufacture a single nanopore on a membrane (30, 41), characterized by comprising: two reservoirs (33) fluidly coupled via a passage (34) to each other, characterized in that the passage is configured to receive a membrane that prevents the liquid from passing between the two reservoirs, where the membrane is comprised of a dielectric material; at least two electrodes (24) electrically connected to a voltage source (25) and operable to generate an electrical potential across the membrane, where an electrode of at least two electrodes is disposed in each of the two reservoirs and the electrical potential induces a electric field that has a value greater than 0.1 volt per nanometer; a current sensor electrically coupled to one of the electrodes and operable to measure the current flowing between the two reservoirs; and a controller (26) which interfaces with the current sensor and detects an abrupt increase in the measured current and, in response to detecting an abrupt increase in the measured current, removes the electrical potential across the membrane.
    [0024]
    Mechanism according to claim 23, characterized in that one of the reservoirs is fluidly separated into two or more micro- or nano-channels, each micro- or nano-channel having an electrode disposed there.
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    Pospelov et al.2015|A new method for controlling the quantized growth of dendritic nanoscale point contacts via switchover and shell effects
    Robinson et al.2017|Electrical and electrochemical characterization of proton transfer at the interface between chitosan and PdH x
    CN108996461A|2018-12-14|A kind of diameter is less than glass nano hole, preparation method and its application for detecting DNA of 10nm
    Hu et al.2014|Detection and analysis of DNA recapture through a solid-state nanopore
    TW201939010A|2019-10-01|Method for detecting interface potential between nanopore and nanoparticle does not require the analysis of a detailed spectrum of graph of the current impedance signal
    EP2908128A1|2015-08-19|Molecular sensing device
    同族专利:
    公开号 | 公开日
    WO2013167952A1|2013-11-14|
    JP6420236B2|2018-11-07|
    EP2847367A1|2015-03-18|
    CA2872600C|2021-11-23|
    US9777389B2|2017-10-03|
    EP2846901B1|2017-03-29|
    WO2013167955A1|2013-11-14|
    CN104662209B|2017-03-15|
    BR112014027873B1|2021-05-25|
    KR20150040799A|2015-04-15|
    CN104662209A|2015-05-27|
    MX2014013412A|2016-04-07|
    US9777390B2|2017-10-03|
    BR112014027829A2|2017-06-27|
    AU2013257759A1|2014-11-27|
    AU2013257756B2|2017-05-04|
    US20150109008A1|2015-04-23|
    MX353370B|2018-01-09|
    SG10201606334XA|2016-09-29|
    EP2847367A4|2016-01-13|
    ES2630064T3|2017-08-17|
    SG11201407249XA|2014-12-30|
    US20150108008A1|2015-04-23|
    KR102065754B1|2020-02-11|
    KR20150037748A|2015-04-08|
    CA2872602C|2020-08-25|
    JP2015525114A|2015-09-03|
    SG11201407252UA|2014-12-30|
    CN104411386B|2016-10-12|
    ES2629952T3|2017-08-16|
    EP2846901A4|2016-01-13|
    CA2872600A1|2013-11-14|
    CN104411386A|2015-03-11|
    AU2013257759B2|2017-07-13|
    BR112014027873A2|2017-06-27|
    MX357200B|2018-06-29|
    EP2847367B1|2017-03-29|
    JP6298450B2|2018-03-20|
    EP2846901A1|2015-03-18|
    CA2872602A1|2013-11-14|
    AU2013257756A1|2014-11-27|
    JP2015517401A|2015-06-22|
    KR102065745B1|2020-01-14|
    JP2018187626A|2018-11-29|
    MX2014013410A|2015-12-08|
    引用文献:
    公开号 | 申请日 | 公开日 | 申请人 | 专利标题
    JPH02173278A|1988-12-26|1990-07-04|Hitachi Ltd|Method and device for fine processing|
    JPH09316692A|1996-05-30|1997-12-09|Fine Ceramics Center|Alumina film having fine pore and its production|
    JP3902883B2|1998-03-27|2007-04-11|キヤノン株式会社|Nanostructure and manufacturing method thereof|
    US7258838B2|1999-06-22|2007-08-21|President And Fellows Of Harvard College|Solid state molecular probe device|
    DE10044565B4|2000-09-08|2005-06-30|Gesellschaft für Schwerionenforschung mbH|An electrolytic cell, its use and method of etching a membrane clamped in the cell, and methods of switching an etched cell clamped membrane from pass-to-pass and vice versa|
    JP2003001462A|2000-09-13|2003-01-08|Hamamatsu Photonics Kk|Laser beam machining device|
    US6592742B2|2001-07-13|2003-07-15|Applied Materials Inc.|Electrochemically assisted chemical polish|
    US6706203B2|2001-10-30|2004-03-16|Agilent Technologies, Inc.|Adjustable nanopore, nanotome, and nanotweezer|
    WO2004077503A2|2003-02-03|2004-09-10|President And Fellows Of Harvard College|Controlled fabrication of gaps in electrically conducting structures|
    JP5138229B2|2003-12-24|2013-02-06|ザリージェンツオブザユニバーシティオブカリフォルニア|Tissue ablation by irreversible electroporation|
    EP1721657A1|2005-05-13|2006-11-15|SONY DEUTSCHLAND GmbH|A method of fabricating a polymeric membrane having at least one pore|
    JP4925670B2|2006-01-16|2012-05-09|埼玉県|Method for manufacturing titanium metal products|
    US7849581B2|2006-05-05|2010-12-14|University Of Utah Research Foundation|Nanopore electrode, nanopore membrane, methods of preparation and surface modification, and use thereof|
    US7777505B2|2006-05-05|2010-08-17|University Of Utah Research Foundation|Nanopore platforms for ion channel recordings and single molecule detection and analysis|
    DE102006035072B4|2006-07-28|2009-03-12|Westfälische Wilhelms-Universität Münster|Device and method for detecting particles with pipette and nanopore|
    WO2008124107A1|2007-04-04|2008-10-16|The Regents Of The University Of California|Compositions, devices, systems, and methods for using a nanopore|
    FR2927169B1|2008-02-05|2013-01-11|Commissariat Energie Atomique|METHOD FOR FUNCTIONALIZING THE SURFACE OF A PORE|
    US20100122907A1|2008-05-06|2010-05-20|Government of the United States of America,|Single molecule mass or size spectrometry in solution using a solitary nanopore|
    EP2237027B1|2009-04-03|2011-11-30|Nxp B.V.|Sensor device and a method of manufacturing the same|
    CA2772789C|2009-09-18|2018-10-30|President And Fellows Of Harvard College|Bare single-layer graphene membrane having a nanopore enabling high-sensitivity molecular detection and analysis|
    CN102781558B|2009-11-25|2015-06-24|悉尼大学|Membrane and membrane separation system|
    DE102010025968B4|2010-07-02|2016-06-02|Schott Ag|Generation of microholes|
    US9422154B2|2010-11-02|2016-08-23|International Business Machines Corporation|Feedback control of dimensions in nanopore and nanofluidic devices|
    KR20120133653A|2011-05-31|2012-12-11|삼성전자주식회사|Nano-sensor, method of manufacturing the same and method of detecting target molecules by using the same|
    JP6437521B2|2013-03-15|2018-12-12|プレジデント アンド フェローズ オブ ハーバード カレッジ|Fabrication of nanopores in atomically thin films by ultrashort electrical pulses|JP6437521B2|2013-03-15|2018-12-12|プレジデント アンド フェローズ オブ ハーバード カレッジ|Fabrication of nanopores in atomically thin films by ultrashort electrical pulses|
    WO2015097765A1|2013-12-25|2015-07-02|株式会社日立製作所|Hole forming method, measuring apparatus, and chip set|
    JP6209122B2|2014-04-02|2017-10-04|株式会社日立ハイテクノロジーズ|Hole forming method and measuring apparatus|
    DE102014111984B3|2014-08-21|2016-01-21|Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V.|Fluidic gigaohm seal for transmembrane protein measurements|
    AU2015361032A1|2014-12-01|2017-06-29|Cornell University|Nanopore-containing substrates with aligned nanoscale electronic elements and methods of making and using same|
    EP3828540A1|2014-12-19|2021-06-02|The University of Ottawa|Integrating nanopore sensors within microfluidic channel arrays using controlled breakdown|
    WO2016135656A1|2015-02-24|2016-09-01|The University Of Ottawa|Localizing nanopore fabrication on a membrane by laser illumination during controlled breakdown|
    CN104694649A|2015-03-10|2015-06-10|北京大学|Nanopore sequencing method for nucleic acid molecules with low perforation rate and special nanopore device thereof|
    EP3268736B1|2015-03-12|2021-08-18|Ecole Polytechnique Fédérale de Lausanne |Nanopore forming method and uses thereof|
    EP3067693A1|2015-03-12|2016-09-14|Ecole Polytechnique Federale de Lausanne |Nanopore forming method and uses thereof|
    WO2017004463A1|2015-07-01|2017-01-05|Abbott Laboratories|Devices and methods for sample analysis|
    JP2018511805A|2015-04-03|2018-04-26|アボット・ラボラトリーズＡｂｂｏｔｔ Ｌａｂｏｒａｔｏｒｉｅｓ|Devices and methods for sample analysis|
    GB201508669D0|2015-05-20|2015-07-01|Oxford Nanopore Tech Ltd|Methods and apparatus for forming apertures in a solid state membrane using dielectric breakdown|
    EP3523640A4|2016-10-05|2020-07-15|Abbott Laboratories|Devices and methods for sample analysis|
    CN109791138B|2016-10-12|2021-01-08|豪夫迈·罗氏有限公司|Nanopore voltage method|
    JP6653767B2|2016-12-09|2020-02-26|株式会社日立ハイテクノロジーズ|Nanopore formation method, nanopore formation device, and biomolecule measurement device|
    WO2018131064A1|2017-01-10|2018-07-19|株式会社日立ハイテクノロジーズ|Current measurement device and current measurement method using nanopore|
    CN110741093A|2017-04-28|2020-01-31|渥太华大学|Controlling translocation of molecules through nanopores|
    US11224842B2|2017-05-17|2022-01-18|The Royal Institution For The Advancement Of Learning / Mcgill University|Method and apparatus for making a nanopore in a membrane using an electric field applied via a conductive tip|
    US10618805B2|2017-09-22|2020-04-14|Applied Materials, Inc.|Method to reduce pore diameter using atomic layer deposition and etching|
    US10830756B2|2017-09-22|2020-11-10|Applied Materials, Inc.|Method to create a free-standing membrane for biological applications|
    US10752496B2|2017-09-22|2020-08-25|Applied Materials, Inc.|Pore formation in a substrate|
    JP6959121B2|2017-12-05|2021-11-02|株式会社日立ハイテク|Hole forming method and hole forming device|
    WO2019109253A1|2017-12-05|2019-06-13|清华大学|Method for adjusting effective size of solid-state nanopore in solid-state nanopore system|
    CN108279312B|2018-03-08|2021-06-01|冯建东|Nanopore-based proteomics analysis device, serum detection method and application|
    JP2020094894A|2018-12-12|2020-06-18|株式会社日立製作所|Nanopore forming method and analysis method|
    WO2021260587A1|2020-06-23|2021-12-30|The University Of Ottawa|Improved techniques for nanopore enlargement and formation|
    WO2022024335A1|2020-07-31|2022-02-03|株式会社日立ハイテク|Biomolecule analysis method, biomolecule analyzing reagent, and biomolecule analysis device|
    法律状态:
    2018-03-06| B06F| Objections, documents and/or translations needed after an examination request according [chapter 6.6 patent gazette]|
    2018-03-13| B06F| Objections, documents and/or translations needed after an examination request according [chapter 6.6 patent gazette]|
    2018-03-20| B06I| Publication of requirement cancelled [chapter 6.9 patent gazette]|Free format text: ANULADA A PUBLICACAO CODIGO 6.6.1 NA RPI NO 2462 DE 13/03/2018 POR TER SIDO INDEVIDA. |
    2019-09-17| B06U| Preliminary requirement: requests with searches performed by other patent offices: procedure suspended [chapter 6.21 patent gazette]|
    2020-09-24| B06A| Patent application procedure suspended [chapter 6.1 patent gazette]|
    2021-02-23| B09A| Decision: intention to grant [chapter 9.1 patent gazette]|
    2021-03-23| 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 07/05/2013, OBSERVADAS AS CONDICOES LEGAIS. |
    优先权:
    申请号 | 申请日 | 专利标题
    US201261643651P| true| 2012-05-07|2012-05-07|
    CA61/643,651|2012-05-07|
    US201361781081P| true| 2013-03-14|2013-03-14|
    CA61/781,081|2013-03-14|
    PCT/IB2013/000891|WO2013167955A1|2012-05-07|2013-05-07|Fabrication of nanopores using high electric fields|
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