• 专利附录
  • 专利说明
  • 权利要求
  • 类似技术
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    • 专利摘要:
    The present invention relates to a system for transferring a fluid sample comprising, first and second fluid circuits connected respectively to a first, to a second input-output (1, 2) of a fluidic cell (10), first injection means (80) adapted to inject in series into the first fluid circuit (11, 12, 13): a buffer solution, a first volume of separation fluid followed by a fluid sample and then a second volume of fluid separation and another buffer solution. According to the invention, the system comprises first discharge means disposed on the first fluid circuit near the first input-output (1), second injection means (90, 91) disposed on the second fluid circuit ( 21, 22, 23) near a second inlet-outlet (2), and bidirectional circulation means adapted to circulate the fluid sample in two opposite directions in the fluidic cell (10) without passage of separation fluid in said fluidic cell (10).
    公开号:FR3017956A1
    申请号:FR1451411
    申请日:2014-02-21
    公开日:2015-08-28
    发明作者:Emmanuel Maillart；Cecile Lerondeau；Geraldine Melizzi；Didier-Luc Brunet；Denis Cattelan
    申请人:Horiba Jobin Yvon SAS；
    IPC主号:
    专利说明:

    [0001] TECHNICAL FIELD TO WHICH THE INVENTION RELATES The present invention relates to a system for transferring a fluid sample to a fluidic cell for the purpose of analyzing this sample and for transferring this fluid sample out of the fluidic cell, after analysis. .
    [0002] The invention applies in particular to apparatus for measuring biological interactions and biosensors. BACKGROUND ART In general, the most widely used fluid sample analysis technique is liquid chromatography (HPLC) in which fluid samples are sent to a column. Other fluid sample analysis systems are provided with a fluidic cell configured for analyzing interactions between the fluid sample and molecules. In these systems, a fluid sample is taken, usually a small amount, the fluid sample is transferred to a fluid cell where an analysis is performed, and then the fluid sample is extracted from the fluid cell. In the present document, the term sample is understood to mean a fluid sample originating from a liquid source (for example a physiological liquid) or from a preparation in solution, for example by dissolving a component in a solvent. Preferably, a fluid cell is configured to allow analysis of the sample by an optical analytical technique. More particularly, the interaction between a sample and a surface of the fluidic cell is analyzed by optical means implementing evanescent optical phenomena such as, for example: surface plasmon resonance or SPR (Surface Plasmon Resonance), but also other types of plasmons such as Localized Surface Plasmon Resonance (LSPR) or Long Range Surface Plasmon Resonance (LRSPR), resonant mirrors, waveguides, or surface Bloch waves ( BSW). Advantageously, the fluidic cell comprises a biochip or a biosensor comprising a functionalized surface to allow interaction and simultaneous analysis of a sample on a matrix of interaction sites, the different interaction sites being dedicated to the interaction with different molecules or particular reagents. In a biosensor, the signal that measures the interaction between an analyte and the biosensor is related to the concentration of the sample to be analyzed and the duration of the interaction. Thus, in the simple case of an interaction between a ligand L immobilized on a biochip and an analyte A in solution that can interact according to a stoichiometry of one to one, the chemical reaction is written: L + A, Aral '` The 'dss OÙ k' represents the association rate constant of the reaction in M-1s-1 and kdi 'the dissociation rate constant of the reaction in s-1.
    [0003] The response R (t) of the biosensor to this reaction can be expressed according to the following equation: R (t) [A]. R. e_ (k ,,,] A] + kd,) t) [A] + KD where Rmax represents the saturation response of the biosensor, KD (= kdissikass) the dissociation constant of the reaction in M, [A] the analyte concentration of the sample and t the time. The higher the concentration of the sample, the higher the measured signal is, as long as the recognition elements of the biochip are not saturated. The longer the passage of the sample in the fluidic cell, the greater the duration of the interaction and the greater the measured signal, as long as the recognition reaction has not reached its equilibrium. It is sought to improve the sensitivity of a fluid analysis system, for example for the analysis of samples in low concentration and / or in small quantities. There are solutions for increasing the passage time of a sample in a fluid cell without increasing the consumption of said sample.
    [0004] One solution is to implement a closed circuit recirculation system, the output of the fluidic cell being connected to the input of the fluidic cell. The advantage of a recirculation in closed circuit, is to avoid the diffusion of the sample regardless of the duration of passage of the sample in the fluid cell. However, the sample volume in recirculation is equal to the volume of the closed circuit. However, for some samples, the available volume can be very small and less than the volume of the closed circuit. For another sample, it may be necessary to circulate in a loop a larger volume of a low concentration sample so that the number of molecules that interacted on the biochip remains negligible compared to the total amount of this molecule in the sample. Thus the concentration of the sample remains constant as the passage time in the fluidic cell, by eliminating the depletion phenomenon. Another solution uses a back-and-forth system, the flow direction of the sample in the fluidic cell being reversed periodically: the output of the fluidic cell periodically becomes the input and vice versa. The back and forth is compatible with different sample volumes, and different sample concentrations.
    [0005] Moreover, the sample to be analyzed generally comprises a solution and target molecules that are to be detected via a biosensor. In general, the sample to be analyzed is injected as a result of a reference liquid of the buffer solution type, such as PBS (Phosphate Buffer Saline) or HBS (HEPES Buffer Solution, with HEPES for 4- (2- hydroxyethyl) -1-piperazine ethane sulfonic acid), not containing target molecules. During the transfer of the sample to the fluid cell, the sample tends to dilute by diffusion in contact with the buffer solution, which decreases its average concentration and therefore the response of the biosensor. Techniques are commonly used to limit the diffusion of a sample during its transfer to a fluid cell. In particular, it is known to place on either side of the sample a bubble of fluid, generally air, immiscible neither with the sample nor with the circulating solution before and after the sample. The sample is thus interposed or framed between an upstream air bubble and an air bubble downstream of the fluidic cell. Each air bubble separates the sample from a reference liquid that circulates in the fluid cell before and after the passage of the sample. Thus, JP2006-242912 discloses a fluid transfer system capable of providing a long reaction time even for a small amount of sample. In particular, JP2006-242912 describes a fluidic system configured for recirculation of the sample in closed circuit, the output of the fluidic cell being connected to the inlet by means of a valve. In another variant, JP2006-242912 describes a fluidic system configured to generate a back-and-forth of the sample in the fluidic cell. In these two configurations, it is also planned to separate the sample to be analyzed from a buffer solution using an air bubble, in order to avoid any mixing between the sample and the buffer solution. In these devices, the air bubbles circulate inside a channel-shaped passage and serve for example to trigger the reversal of the direction of movement in a movement back and forth. In the fluid cell, the sample is analyzed by means of an optical analytical technique based, non-exhaustively, on evanescent optical phenomena such as SPR, but also other types of plasmons such as localized plasmons (LSPRs). ) or long-distance plasmons (LRSPRs), resonant mirrors, or surface Bloch waves (BSW). In JP2006-242912, which refers to patent document JP3294605 for the structure of the SPR measuring equipment, the fluidic analysis cell has a channel shape, which has a shape ratio, defined as the ratio between width and height, close to 1. This channel can be likened to a tube. But a channel is not adapted to contain a biochip. In this document, the fluidic system is used both for the functionalization of the channel, that is to say the immobilization of the recognition probe molecules, and for the injection of the sample intended to be possibly recognized by the molecules. probes. The arrival of an air bubble in a channel is not a problem because this bubble fills the entire section of the channel and the reference liquid pushes the bubble towards the exit. In addition, the functionalization of the channel being carried out via the fluidic system, allows a homogeneous functionalization of the whole channel and therefore a constant hydrophobicity and / or hydrophilicity. There is therefore no area likely to trap a bubble in this channel. However, a fluidic cell is configured to immobilize several or even several tens or even hundreds of different ligands or recognition elements or probes. For this purpose, the fluidic analysis cell generally has an enlarged shape and has a form factor much greater than 1, with for example a width of several millimeters and a height of several tens of microns. This type of fluidic analysis cell of enlarged section is commonly used in devices for imaging a biochip composed of a matrix of pads (microarray), each pad may contain a different probe molecule immobilized on said chip. In the remainder of this document, we denote by channel a fluidic analysis cell having a form factor -1 and fluidic cell only the cells of fluidic analysis of form factor "1. In the case of the use of a fluidic cell, the fluid (liquid or gas) in circulation will pass over the biochip and thus be in contact with, depending on the locations of the biochip, the surface chemistry or the one or other of the different immobilized probe molecules. These different chemical and / or biological compounds immobilized on the surface have different hydrophobicities. Thus, a preferred direction of passage of an air bubble in the fluidic cell is formed. Areas likely to trap a portion of the air bubble in the fluid cell may also be formed. For example, the patent WO2012 / 045325 describes an SPR (Surface Plasmon Resonance) measurement system and more particularly the introduction of a back and forth in the fluidic system to increase the reaction time without increasing the consumption of reagents. W02012 / 045325 mentions that to do this back and forth, it is essential to separate the buffer solution and the sample by a fluid of very different refractive index such as an immiscible liquid or a gas such as air. This separation prevents buffer and sample from mixing by diffusion. According to this document WO2012 / 045325, the passage of an air bubble in the fluidic cell is used to trigger the beginning and the end of the measurement of the sample by detecting a sudden resonance shift induced by the sudden jump. of refractive index between the liquid medium and the air bubble. However, an air bubble is likely to adhere to the sensitive surface of a sensor in a fluidic cell and to modify its surface properties, in particular when a hydrophobic layer is deposited on the sensor (see Handbook of Surface Plasmon Resonance, chapter 3.3.1 page 46). It is known to use an in-line degasser to prevent the formation of bubbles by degassing the sample or buffer solution in the fluid cell. It therefore seems equally important to prevent an air bubble used to separate the sample from the buffer solution from being trapped inside a fluid cell. Thus, WO00 / 025547 discloses a system and method for analyzing a surface plasmon resonance sample in a microfluidic cell, and provides capillaries and zones for trapping an air bubble separating the sample. buffer solution before the air bubble enters the fluid cell. More specifically, the device comprises a fluid circuit for bringing the sample in solution to at least one reaction chamber to interact with one or more reagents. The fluid circuit comprises one or more vents for removing air bubbles from the fluidic cell or the fluid circuit upstream of the fluidic cell.
    [0006] However, this device allows to circulate the sample in one direction, and does not make it possible to go back and forth from the sample in the analysis chamber of the fluidic cell. There is therefore a need for a system and a method for increasing the interaction time of a fluid sample in a fluidic cell, while avoiding diffusion between the fluid sample and a circulating fluid, such as than a buffer solution, and while avoiding modifying the interaction surface of the fluidic cell due to the passage and / or trapping of an air bubble in the fluid cell. The present invention therefore proposes to solve the problem of increasing the passage time of a sample in a fluidic cell, in particular having a form factor much greater than 1, compatible with different sample volumes, without increasing the consumption, and while limiting as much as possible the diffusion of the sample towards other liquids, such as a buffer solution, during the passage of this sample in the fluidic circuit, and while avoiding modifying a surface of interaction of the fluidic cell during the passage of a separation fluid, for example an air bubble, in the fluid cell. OBJECT OF THE INVENTION The object of the present invention is to overcome the drawbacks of prior systems and more specifically proposes a system for transferring a fluid sample into a fluidic cell, the fluidic system comprising a fluidic cell having a first input-output arranged. upstream of said fluidic cell and a second input-output disposed downstream of said fluidic cell, a first fluidic circuit connected to said first input-output and a second fluidic circuit connected to said second input-output, first injection means connected to the first fluid circuit, said first injection means being adapted to inject in series into the first fluid circuit: a buffer solution, a first volume of separation fluid followed by a fluid sample and then a second volume of fluid; separation and another buffer solution, and circulation means adapted to make circulating said fluid sample interposed between the first volume of separation fluid and the second volume of separation fluid in the first fluid circuit towards the first input-output of the fluid cell. According to the invention, the system comprises first evacuation means disposed on the first fluid circuit near the first input-output, said first evacuation means being adapted to extract, upstream of the fluidic cell, said first volume separating fluid and, respectively, said second volume of separation fluid, second injection means disposed on the second fluid circuit in the vicinity of the second input-output, said second injection means being adapted to inject, downstream of the fluidic cell, a third volume of separation fluid between said buffer solution and the fluid sample and, respectively, a fourth volume of separation fluid between the fluid sample and said other buffer solution, and suitable bidirectional circulation means. to circulate the fluidic sample in a first direction of circulation from lad said first input-output to said second input-output or a second direction of flow from said second input-output to said first input-output without passing the first, second, third or fourth volume of separation fluid into said fluid cell . The system of the invention, integrated on both sides of the fluidic cell, makes it possible to transfer a fluid sample, to evacuate the first separation bubble preceding the sample before it enters the fluidic cell and to reinject a new one. separation bubble just after the exit of the sample downstream of the fluidic cell.
    [0007] This system can be used in the same way to remove and reinject the second air bubble following the sample. Thus, each end of the sample is in contact with the buffer solution only during the duration of a passage through the fluid circuit between the first discharge means and the second injection means of a separation bubble, and regardless of the length of the fluid circuit upstream of the fluid cell and regardless of the total sample passage time in the fluid cell. The duration of contact between the sample and the buffer solution and the induced diffusion are shorter as the first evacuation means and the second injection means are located closer to the fluidic cell. Nevertheless, this time remains slightly greater than the transit time of the sample through the fluidic cell.
    [0008] The system and method of the invention allow a reciprocating flow of the sample in the fluidic cell during the time required for interaction with a sensitive surface of a sensor, while introducing a diffusion and therefore a dilution of the minimum sample. Advantageously, the system comprises an optical analysis device coupled to the fluidic cell and configured to allow an optical analysis, for example of the SPR or LSPR ... type, of the sample in the fluidic cell, or of an interaction between the sample and the sensitive surface of a sensor. According to a first embodiment, the first evacuation means comprise a first valve having at least a first input-output port fluidly connected to the first input-output of the fluidic cell, a second connected input-output port. fluidically to the first injection means, and a third input-output port, said first valve having at least one first state, wherein the first input-output port of the first valve is connected to the second input port output of the first valve and a second state, wherein the second input-output port of the first valve is connected to the third input-output port of the first valve. According to a particular aspect of the first embodiment, the second separation fluid injection means comprise a second valve having at least a first input-output port fluidly connected to the second input-output of the fluidic cell, a second an input-output port and a third input-output port fluidly connected to a source of separation fluid, said second valve having at least one first state, wherein the first input-output port of the second valve is connected to the second input-output port of the second valve and a second state, wherein the second input-output port of the second valve is connected to the third input-output port of the second valve. According to a second embodiment, the first means for evacuating the separation fluid and the second separation fluid injection means comprise a valve having at least a first input-output port fluidly connected to the first input-output of the fluidic cell, a second input-output port fluidically connected to the second input-output of the fluidic cell, a third input-output port connected to the first injection means, a fourth input-output port fluidly connected to a source of separation fluid, a fifth input-output port and a sixth input-output port, said fifth and sixth input-output ports preferably being fluidly connected to an evacuation device or recovery device, said valve having at least a first state in which the first input-output port is connected to the third input-output port and in which the second input-output port is output is connected to the sixth input-output port, said valve having at least one second state in which the third input-output port is connected to the fifth input-output port and / or in which the fourth port of input-output is connected to the sixth input-output port.
    [0009] According to a first variant of the system of the invention, the system comprises a detector disposed on the first fluid circuit between the first injection means and the first means for discharging the separation fluid, the detector being adapted to detect the first volume of separation fluid and / or the second volume of separating fluid. According to a particular aspect of the invention, the third volume of separation fluid being a volume of air, the second separation fluid injection means comprise an air pump preferably connected to an air filter, the third and the fourth volume of separating fluid being a volume of air. According to a particular and advantageous aspect of the invention, the fluidic cell has a shape ratio greater than the aspect ratio of the first and second fluid circuits respectively. In a particularly advantageous manner, the fluidic cell comprises a biochip adapted for the analysis of the fluid sample in at least one analysis site, and preferably in a matrix of analysis sites, the analysis preferably being an analysis. such as surface plasmon resonance (SPR) resonance, localized surface plasmon resonance (LSPR), resonant mirror, surface Bloch wave (BSW), integrated waveguides or resonant microcavities (WGM). According to a particular and advantageous aspect of the invention, the system comprises synchronization means adapted to synchronize the operation of the first separation fluid evacuation means, the second separation fluid injection means and bidirectional circulation means. . Advantageously, the fluidic system further comprises a reservoir adapted to receive the fluid or fluids from the second input-output of the fluidic cell.
    [0010] The invention also relates to a method for transferring a fluid sample in a fluidic system, the method comprising the following steps: series injection into a first fluid circuit of a buffer solution followed by a first volume of separation fluid, of a fluid sample, then a second volume of separation fluid and another buffer solution, circulation of the fluid sample interposed between the first volume of separation fluid and the second volume of separation fluid in the first fluid circuit in the direction of the first input-output of the fluidic cell, evacuation, near the first input-output, of the first volume of separation fluid, upstream of the fluidic cell, passage of the buffer solution followed by fluid sample in the fluid cell; injecting, near the second input-output, a third volume of separation fluid between said buffer solution and the fluid sample downstream of the fluidic cell, and bi-directional circulation of the fluid sample inserted between the second separating fluid volume and the third separation fluid volume, in a first direction of flow from said first input-output to said second input-output and / or a second direction of flow from said second input-output to said first input-output without passing the second or third volume of separation fluid into said fluid cell; evacuation, near the first inlet-outlet, of the second volume of separation fluid, upstream of the fluidic cell, passage of the fluid sample in the fluidic cell followed by the other buffer solution through the fluid cell of the first input-output to the second input-output; injecting, near the second input-output, a fourth volume of separation fluid between the fluid sample and the other buffer solution downstream of the fluidic cell. The present invention also relates to the features which will emerge in the course of the description which follows and which will have to be considered individually or in all their technically possible combinations.
    [0011] This description, given by way of nonlimiting example, will make it easier to understand how the invention may be made with reference to the appended drawings, in which: FIGS. 1A-1 F schematically represent the main steps of an example of a transfer method according to FIG. invention; FIG. 2 illustrates an example of a numerical simulation of interaction kinetics of a sample with a recognition element immobilized on the chip and directed against one of the constituents of the sample; FIGS. 3A-3C show schematically a transfer system of a fluid sample according to a first embodiment of the invention; FIGS. 4A-4B schematically represent a system for transferring a fluid sample according to a second embodiment of the invention; FIG. 5 diagrammatically represents a first variant of a fluid sample transfer system according to the first embodiment of the invention; FIG. 6 schematically represents a first variant of a fluid sample transfer system according to the second embodiment of the invention; FIG. 7 schematically represents a second variant of a fluid sample transfer system according to the first embodiment of the invention; FIG. 8 schematically represents a second variant of a transfer system of a fluidic sample according to the second embodiment of the invention; FIG. 9 schematically represents a third variant of a fluid sample transfer system according to the first embodiment of the invention; FIG. 10 diagrammatically represents a third variant of a fluid sample transfer system according to the second embodiment of the invention; - Figure 11 schematically shows a complete system for transferring a fluid sample according to one embodiment of the invention. FIGS. 1A-1F schematically illustrate the principle of a system according to one embodiment of the invention and the main steps of the transfer method. Systems and Methods In FIGS. 1A-1F, there is shown a fluidic cell 10 for receiving a sample in liquid or disperse solution form which it is desired to analyze. Figures 1- A to 1-F schematically show a fluid cell 10 in plan view. In the plane of Figures 1-A to 1-F, this fluidic cell 10 has a hexagonal general shape, or square, or rectangular, or other. In a plane transverse to the plane of FIGS. 1-A to 1-F, the fluidic cell 10 has a section of rectangular and elongated shape. The fluidic cell 10 comprises a first input-output 1 disposed upstream of the fluidic cell 10 and a second input-output 2 disposed downstream of said fluidic cell 10. The two input-outputs 1,2 are for example disposed at the opposite vertices. of a hexagon. The first input-output 1 is fluidly connected to a first fluid circuit, of which two branches 12 and 13 have been represented. Similarly, the second input-output 2 is fluidly connected to a second fluid circuit, of which two branches 22 have been represented. and 23. The branches 12 and 13 are for example made of metal tubes or polymer material. The fluidic cell 10 is generally coupled to an analysis system, for example an optical analysis system, of the SPR, LSPR, LRSPR or BSW type. Advantageously, the fluidic cell is in the form of a trough. , adapted to receive a fluid sample. By way of example, the fluidic cell can be closed in a sealed manner by an optical coupling interface on which an optical beam is incident, so as to measure the interactions between this optical beam and the fluid sample. Particularly advantageously, the optical coupling interface comprises a biosensor or a biochip on the surface in contact with the fluid sample. A biochip generally comprises different molecules immobilized on a surface and arranged in a matrix. In order to make it possible to immobilize a large quantity of ligands, or different recognition elements or probes, the fluidic cell 10 preferably has a form factor "1, typically with a width of several millimeters and a height of a few tens of microns. . In general, an injection and circulation system, such as an auto-injector, described in detail in connection with FIG. 11, draws a fluid sample 30 and injects it into a branch 13 of the first fluid circuit. The injection and circulation system advances the fluid sample into the fluid cell where it interacts with a surface for analysis. The fluid sample 30 is then transferred to the second fluid circuit for disposal or collection. The flow direction in the fluidic circuit is indicated by an arrow in FIGS. 1A, 1-B, 1-C, 1-E and 1-F. To illustrate a reciprocating circulation, two opposite arrows in FIG. 1-D are shown. In general, the auto-injector first circulates a reference liquid 35, or buffer solution, generally containing a solvent, such as water, or water and salts, for example PBS or HBS, through the fluidic circuit 13, 23 and the fluidic cell 10. Unlike the fluid sample 30 that is to be analyzed via the biosensor, the buffer solution 35 does not contain target molecules. The buffer solution 35 makes it possible to keep the fluid circuit 13, 23 and the fluidic cell 10 clean, and avoids any contamination of the biosensor before the fluid sample 30 enters the fluidic cell 10. The signal measured during the passage of the Buffer solution 35, before the sample, serves as a reference signal. The signal measured during the passage of the buffer solution 36 after the sample is used to measure the dissociation, that is to say the speed of separation between the probes and the targets. To avoid dilution of the fluid sample 30 in the buffer solution 35 when it is transferred to the fluidic cell 10, the auto-injector inserts, just before and immediately after the fluid sample 30, a separation fluid bubble. The separation fluid is selected to be immiscible with neither the fluid sample nor the buffer solution. The separation fluid is generally air or an inert gas such as nitrogen or argon.
    [0012] Thus, FIG. 1-A shows the passage of a buffer solution 35 through the fluidic cell 10, from the first input-output 1 to the second input-output 2, in the direction of the branch 23 of the second fluid circuit. . A first air bubble 31 separates the buffer solution 35 from the sample 30. The first air bubble 31 and the fluid sample 30 circulate in the branch 13 of the first fluid circuit towards the first inlet-outlet of the fluidic cell 10. A second air bubble 32 separates the sample 30 from the injected buffer solution 36 following the sample in the first fluid circuit (see Fig. 1-C and 1-D). Thus, the sample 30 flows in the branch 13 of the first fluid circuit upstream of the fluidic cell 10, the sample 30 being interposed between the first air bubble 31 and the second air bubble 32. In FIG. -A, the branch 12 of the first fluid circuit and, respectively the branch 22, of the second fluid circuit, are hermetically closed by a valve 41, respectively 42. The sample 30 preceded by the first air bubble 31 is injected up to near the first input-output 1 of the fluidic cell 10. In FIG. 1-B, the valve 41 on the branch 12 of the first fluid circuit is open and a valve 43 hermetically closes the circuit between the branch 13 and the first input-output 1 of the fluidic cell 10. The auto-injector induces a circulation of the first air bubble 31 to the branch 12 of the first fluid circuit. In the fluidic cell 10, the circulation of the buffer solution 35 is stopped. Thus, the first air bubble 31 is evacuated before entering the fluidic cell 10. Indeed, an air bubble entering the fluidic cell 10 is likely to be fixed and modify the contact interface between the sample to be analyzed 30 and the interaction surface, for example a biosensor. In addition, it is generally difficult to evacuate an air bubble trapped in the fluidic cell 10. As soon as the first air bubble 31 is extracted via the branch 12 of the first fluid circuit, valve 41 is closed (see FIG. Fig. 1-C) and the valve 43 is reopened to limit a loss of volume of the fluid sample 30 to be analyzed. The auto-injector then induces a flow of the fluid sample 30 towards the fluidic cell. During the injection of the fluid sample 30 from the first inlet-outlet 1 to the second inlet-outlet, no separation fluid separates the fluid sample 30 from the buffer solution 35 which has already passed through the fluidic cell. However, this step without separating fluid is limited to the duration of a passage of the sample in the cell 10 and the resulting dilution of the sample is generally very small. In FIG. 1-C, the buffer solution 35 is taken out of the fluidic cell 10 and the sample 30 reaches the second input-output 2 of the fluidic cell 10. A valve 44 is closed near the second input-output 2 and opening a valve 42 on a branch 22 of the second fluid circuit. In the fluidic cell 10, the circulation of the sample 30 is stopped. A new air bubble 33 is inserted at the interface between the sample 30 and the buffer solution 35. The new air bubble 33 is inserted outside the fluidic cell 10.
    [0013] The injection system induces a circulation of the new air bubble 33 and the buffer solution 35 to the branch 23 of the second fluid circuit. A second air bubble 32 separates the sample 30 from another separation fluid 36, which is generally of the same kind as the first separation fluid 35. The second air bubble 32 and the other separation fluid 36 are located on the branch 13 of the first fluid circuit upstream of the first inlet-outlet 1 of the fluidic cell 10. Thus, in FIG. 1-D, a fluidic sample 30 is obtained which fills the fluidic cell 10, the fluid sample 30 being interposed between two air bubbles 33, 32 separating it from the buffer solution 35, 36, without any air bubble being injected into the fluidic cell 10. An air bubble 32 is located on the branch 13 of the first fluid circuit upstream of the first input-output 1 of the fluidic cell 10. And another air bubble 33 is located on the branch 23 of the second fluid circuit downstream of the first input-output 1 of the fluidic cell 10. Advantageously, the autoinjector induces an alternating bidirectional circulation, in other words a reciprocating circulation, of the sample 30 in the fluidic cell 10, the two air bubbles 32, 33 which surround the sample remaining outside the fluidic cell 10. Preferably, the fluid sample 30 has a volume greater than the internal volume of the fluidic cell 10. The volume of the fluid sample is determined so that the displacement of the sample induced by a movement of back and forth is interrupted before the entry of the air bubble 32 via the first input-output 1 and before the entry of the air bubble 33 via the second input-output 2. The circulation in va- and forth increases the interaction time and the probability of interaction between the fluid sample and the surface of a biosensor in the fluid cell.
    [0014] During the reciprocating circulation, the sample is separated from the buffer solution 35, 36 at both ends, which makes it possible to avoid any dilution of the sample during the back and forth. FIGS. 1 -E and 1-F illustrate the extraction of the sample 30 from the fluidic cell 10. The auto-injector induces a circulation of the sample 30 from the first input-output 1 to the second input- outlet 2 of the fluidic cell 10. When the air bubble 32, which separates the sample from the buffer solution 36 in the first fluid circuit, approaches the first inlet-outlet 1 of the fluidic cell 10, the valve is closed 43 and opens the valve 41 to the branch 12. The auto-injector and pushes the air bubble 32 to the branch 12 of the first fluid circuit. Thus, the air bubble 32 is discharged before entering the fluid cell 10. In the fluid cell 10, the flow of the sample 30 is stopped. As soon as the air bubble 32 is extracted via the branch 12 of the first fluid circuit, the valve 41 is closed again and the valve 43 is opened. The auto-injector again induces a circulation of the fluid sample 30 and the buffer solution 36 through the fluidic cell 10 from the input-output 1 to the input-output 2.
    [0015] During the extraction of the fluid sample from the fluidic cell after evacuation of the air bubble 32, no separating fluid separates the fluid sample 30 from the buffer solution 36. However, this step is limited to the duration of a passage of the sample in the cell 10 and the dilution of the sample resulting therefrom is generally very small. In FIG. 1-F, the sample 30 is extracted from the fluidic cell 10 and the buffer solution 36 reaches the second input-output 2 of the fluidic cell 10. The valve 44 is closed near the second input-output 2 and the valve 42 is opened on the branch 22 of the second fluid circuit. In the fluidic cell 10, the circulation of the buffer solution 36 is stopped. A new air bubble 34 is inserted at the interface between the sample 30 and the buffer solution 36. Thus, an air bubble 34 separates the sample 30 from the separation fluid 36, this new air bubble 34 being inserted outside the fluidic cell 10. The injection system induces a circulation of the new air bubble 34 and the sample 30 to the branch 23 of the second fluid circuit. The air bubble 34 is injected on the branch 23 of the second fluid circuit downstream of the second inlet-outlet 2 of the fluidic cell 10. On the second fluidic circuit, the sample 30 circulates again flanked by two bubbles. The sample 30 can then be collected for further analysis, without having undergone significant dilution in the buffer solution 35 or 36. Thus, during the steps 1-A to 1-F no bubbles separation device 31, 32, 33, 34 has not been injected into the fluidic cell 30. Nevertheless, throughout the reciprocating step, the system and the method allow for a circulation in circulation. from the sample 30 into the fluid cell, the sample remaining separated from the buffer solution 35, 36 by an air bubble 32, 33 respectively disposed at each of its ends, as shown in FIG. 1-D .
    [0016] In an exemplary application, the operator only has 500 μL of a biological sample comprising a target molecule at a concentration of 500 μM. A biosensor is functionalized with a recognition molecule having an affinity Kd = 50nM with the target molecule (ka = 1 -104 M-1s-1 and kd = 5.10-4 s-1). The kinetics of interaction according to a simple 1: 1 stoichiometric model generally follows a curve as indicated in FIG. 2, with an increasing phase as a function of time until reaching equilibrium. The decreasing phase of the curve in FIG. 2 corresponds to the dissociation phase: after the end of the sample has passed into the fluidic cell (here at t = 9000s), the buffer solution 36 circulates again in the fluidic cell . The concentration of analyte therefore goes to 0, which displaces the equilibrium. Recognized molecules will gradually become detached at a more or less sustained rate. In a system where the flow of the sample in the fluidic cell is unidirectional, and for a flow rate of 100g / min, the interaction time of the sample is then limited to 300 s maximum, which does not allow not to reach the equilibrium of the reaction, but only about 10% of the maximum value in FIG. 2. On the contrary, with the same sample and the system and the method of the invention, a bidirectional circulation of the sample for a fortnight back and forth in the fluid cell achieves a total interaction time of 2:30, and the equilibrium of the reaction. This results in a final response 10 times greater than with a fluid system in which the circulation is monodirectional, although the total amount of available sample is extremely limited. During the back and forth, the sample remains separated from the buffer solution, which avoids any dilution of the sample during these many back and forth. The system and method described in connection with FIGS. 1-A to 1-F can be fully automated.
    [0017] FIGS. 3A-3C show a system for transferring a fluid sample according to a first embodiment of the invention. The system of FIGS. 3A-3C comprises an auto-injector 80, a fluidic cell 10, an air pump 91 provided with an air filter 90. The first input-output 1 of the fluidic cell 10 is connected to a first branch 11 of the first fluid circuit. The second input-output 2 of the fluidic cell 10 is connected to a first branch 21 of the second fluid circuit. A first valve 50 is disposed on the first fluid circuit near the first inlet-outlet 1. Advantageously, the first valve 50 comprises at least one inlet 52 and two outlets 51, 53. The inlet 52 of the first valve 50 is connected by a branch 13 to the auto-injector 80. The output 51 of the first valve 50 is connected by the branch 11 to the first input-output 1 of the fluidic cell 10. The output 53 of the first valve 50 is connected by a branch 12 to a container 81, which serves for example as a waste collector. A second valve 60 is disposed on the second fluid circuit near the second inlet-outlet 2 of the fluidic cell. Advantageously, the second valve 60 comprises at least two inputs 61, 63 and an output 62. The input 61 of the second valve 60 is connected by a branch 21 to the second input-output 2 of the fluidic cell 10. The input 63 of the second valve 60 is connected by the branch 22 to the air pump 91. The outlet 62 of the second valve 60 is connected by a branch 23 to a container 93, which serves for example as a collector for recovering the sample after analysis or trash. In FIG. 3-A, the inlet 52 and the outlet 51 of the first valve 50 are fluidly connected. The outlet 53 of the first valve 50 is not connected to the inlet 52, so that the branch 12 is isolated from the first fluid circuit. On the second fluid circuit, the inlet 61 and the outlet 62 of the second valve 60 are fluidly connected. The inlet 63 of the second valve 60 is not connected to the outlet 62, so that the branch 22 is isolated from the second fluid circuit. The autoinjector 80 is used to inject automatically in the branch 13 of the first fluid circuit a framed sample of two air bubbles. The autoinjector 80 can then circulate the sample between two air bubbles in the direction from the autoinjector 80 to the container 93 or in the opposite direction.
    [0018] FIG. 3-A makes it possible to illustrate the step of injecting the sample into the fluidic cell, or the step of going back and forth of the sample in the fluidic cell, or else the step of extracting the sample out of the fluid cell after analysis. In FIG. 3-B, the first valve 50 has been switched, the second valve being in the same position as in FIG. 3-A. On the first fluid circuit, the inlet 52 and the outlet 53 of the first valve 50 are fluidly connected. The outlet 51 of the first valve 50 is not connected to the inlet 52, so that the branch 11 is isolated from the first fluid circuit. The autoinjector 80 directs the fluid via the branches 13 and 12 to the container 81. Figure 3-B shows the configuration of a system for extracting an air bubble just before the bubble enters the fluid cell through the first input-output 1. Figure 3- B illustrates the step of extracting an air bubble 31 which precedes the sample to be analyzed just before the sample enters the fluid cell. FIG. 3-B also illustrates the step of extracting an air bubble 32 which follows the sample, during the step of extracting the sample from the fluidic cell.
    [0019] In FIG. 3-C, the first valve 50 and the second valve 60 have been switched with respect to FIG. 3-B. The first valve 50 is in the same position as in Figure 3-A. On the second fluid circuit, the inlet 63 and the outlet 62 of the second valve 60 are fluidly connected. The inlet 61 of the second valve 60 is not connected to the outlet 62, so that the branch 21 is isolated from the second fluid circuit. The air pump 91 induces fluid flow via the branches 22 and 23 to the container 93. Figure 3-C shows the configuration of a system for inserting an air bubble just after the second input-output 2 of the cell fluidics. FIG. 3-C illustrates the step of injecting an air bubble 33 which precedes the sample to be analyzed, when the sample 30 is in the fluid cell 10. FIG. 3-C also illustrates the step injection of an air bubble 34 which follows the sample, just after the step of extracting the sample out of the fluidic cell. A method of transferring a fluid sample into a system according to the first embodiment comprises the following steps: a) the autoinjector 80 draws an air bubble 32, b) the autoinjector 80 draws the sample 30, c ) the autoinjector 80 takes another air bubble 31, d) the autoinjector 80 injects into the first fluid circuit the sample 30 interposed between the two air bubbles 31, 32, for example in the order 31- 30-32, f) the autoinjector moves the fluids in the first fluid circuit to the fluidic cell 10 and towards the container 93, e) switching the first valve 50 to discharge the bubbles to the container 81 'just before the arrival of the first bubble 31 at the first valve 50, f) evacuation of the first bubble 31 to the container 81, preferably with a margin of safety, including a little buffer solution 35 located just before the first bubble 31, the first bubble 31st a small volume of sample 30 just after the first bubble 31, g) switching of the first bubble evacuation valve 50 to the fluidic cell 10, h) arrival of the beginning of the sample 30 in the fluid cell 10, i) switching of the second bubble reinjection valve 60, just before the arrival of the beginning of the sample 30 at this second valve 60, which then passes from the fluidic cell inlet 10 to the air pump inlet 91, j) starting the air pump 91 to generate an air bubble 33 and injecting this air bubble 33 into the second fluid circuit between the second valve 60 and the container 93. k) tilting of the second valve 60 for reinjecting the bubbles towards the second input-output of the fluidic cell 10 so that the sample 30 is preceded by the bubble 33; I) inversion of the flow direction of the fluids in the fluid circuit just before the arrival of the second bubble 32 at the level of the a first bubble discharge valve 50, the sample 30 flowing in the direction of the container 93 to the fluid cell 10 and the autoinjector 80; m) inversion of the flow direction of the fluids in the fluid circuit, just before the arrival of the bubble 33 at the second bubble reinjection valve 60, the sample 30 then flows in the direction of the autoinjector 80 to the fluidic cell 10 and the container 93; n) repeating the two preceding steps a sufficient number of times to reach the required interaction time, that is to say the number of passages of the sample in the fluidic cell, o) switching of the first valve 50 d evacuation of bubbles, just before the arrival of the second bubble 32 at the first valve 50 to the trash 81, p) discharge to the trash 81 of the second bubble 32, preferably with a safety margin, including a small volume of sample 30 just before the bubble 32, the bubble 32 and a little buffer solution 36 located just after the bubble 32, q) switching of the first valve 50 to discharge the bubbles to the fluidic cell 10, r ) end of the passage of the sample 30 in the fluidic cell 10, s) switching of the second valve 60 of bubble reinjection, just before the arrival of the end of the sample 30 at the valve 60, which passes then from the second input-output 2 of the fluidic cell 10 at the air pump inlet 91, t) actuating the air pump 91 to generate an air bubble 34 and introducing it into the second fluid circuit between the bubble generation valve 60 and the container 93, u) tilting of the second bubble reinjection valve 60 to the second inlet-outlet 2 of the fluidic cell 10 so that the sample 30 is followed by the bubble 34. As a variant, the step j) is replaced by the following step: j ') starting the air pump 91 for injecting air into the fluid circuit between the second bubble generation valve 60 and the container 93, which is equivalent to the generation of A bubble 33. FIGS. 4A-4B show a system for transferring a fluid sample according to a second embodiment of the invention. The same elements are indicated by the same reference signs as in FIGS. 3A-3C. The first valve 50 and the second valve 60 are here replaced by a single valve 70 having six input-outputs 71, 72, 73, 74, 75, 76. The input-output 71 is fluidly connected by a branch 11 to the first input-output 1 of the fluidic cell 10; the input-output 72 is fluidly connected by a branch 21 to the second input-output 2 of the fluidic cell 10; the input-output 73 is fluidly connected by a branch 13 to the auto-injector 80; the inlet-outlet 75 is fluidly connected by a branch 12 to a collecting container 81; the inlet-outlet 74 is fluidly connected by a branch 23 to another collecting container 93; the inlet-outlet 76 is fluidly connected by a branch 22 to an air pump 91 and an air filter 90.
    [0020] The valve 70 is preferably a valve with two operating states. Figure 4-A illustrates the configuration of the transfer system of a fluid sample, the valve 70 being in a first state. In the first state, the input-output 73 is fluidly connected to the input-output 71. Thus, the fluid from the autoinjector 80, connected to the input-output 73, can be directed to the input 1 of the fluidic cell 10 connected to the input-output 71. On the other hand, in the first state, the input-output 72 is fluidly connected to the input-output 74. Thus, the fluid coming from the fluidic cell 10, connected to the output 2 of the fluidic cell 10, can be directed to the collection container 93 connected to the input-output 74. FIG. 4A represents the fluid circuit in its main state, or first state, during the passage of the sample in the fluidic cell 10. In this first state, the autoinjector 80 can either push the sample to the container 93 along the path formed by: the branch 13, the input-output 73, the input-output 71, the branch 11, the input-output 1, the fluidic cell 10, the input- output 2, the branch 21, the input-output 72, the input-output 74 and the branch 23. In this first state, the autoinjector 80 can also suck the sample in the opposite way. The auto-injector 80 can therefore drive a flow of the fluid sample back and forth, in other words back and forth. The inputs-outputs 75 and 76 of the valve V3 being connected, it is possible to send air to the container 81 along the path from the air filter 90, via the pump 91, the branch 22, then to the input-output 76, the input-output 75, the branch 12, to the container 81. This air supply directly to the container 81 does not constitute a step of the method of the invention. In the first state of the valve 70, it is preferable to turn off the air pump 91. In summary, FIG. 4-A illustrates the operation of the system according to the second embodiment, during the injection step of FIG. the sample in the fluidic cell, during the step of expelling the sample out of the fluidic cell, or during a reciprocating flow of the sample in the fluid cell.
    [0021] Figure 4-B illustrates the configuration of the transfer system of a fluid sample, the valve 70 being in a second state. In the second state, the input-output 73 is fluidly connected to the input-output 75. In the second state, the fluid from the autoinjector 80 can be directed to the collection container 81 connected to the input-output. output 75 (Fig. 4-B). Thus, an air bubble 31, or 32, from the autoinjector 80, connected to the input-output 73, can be directed to the collection container 81 connected to the input-output 75. On the other hand in the second state, the input-output 74 is fluidly connected to the input-output 76. Thus, an air bubble 33, or 34, from the air pump 91, connected to the input- 76, can be directed to the branch 23 of the fluid circuit connected to the input-output 74. Figure 4-B illustrates the operation of the system according to the second embodiment, during the step of extracting a air bubble, upstream of the fluidic cell or during the step of injecting an air bubble 33, or 34, downstream of the fluidic cell. FIG. 4B shows the fluidic circuit in a second state which allows the manipulation of the bubbles around the fluidic cell 10. In this second state, if we want to evacuate an air bubble (31 or 32) upstream of the fluidic cell 10 , the autoinjector 80 can push the fluid and in particular an air bubble toward the container 81 along the path from the branch 13, to the input-output 73, the input-output 75, the branch 12 to the container 81, while the air pump 91 is stopped so as not to inject another air bubble undesirably. By cons, if we want to reinject an air bubble (33 or 34) downstream of the fluid cell 10, the air pump 91 can then be turned on. The air pump 91 draws air through the filter 90 and pushes the sucked air to the container 93 along the path from the branch 22, to the input-output 76, the input-output 74, and the branch 23, towards the container 93, while the autoinjector 80 ceases to push the fluids to avoid sending them directly to the container 81 without passing through the fluidic cell 10.
    [0022] The second embodiment has the advantage of using a single valve instead of two valves and therefore to be more compact and less expensive. Thus, in the second embodiment, the valve 70 being in the second state, the input-output 71 is fluidly connected to the input-output 72. In a variant, by inserting an additional pump on the branch 11 or 21 , the sample can be circulated in the closed circuit fluidic cell, from the first inlet-outlet 1 of the fluidic cell to the second inlet-outlet 2 of the fluidic cell, passing through the branches 11 and 12 and the inlet-ports. 71, 72 outlets of the valve 70. Additional fluid flow means are arranged to allow to drive a unidirectional or bidirectional movement to the sample 30 in this recirculation loop.
    [0023] FIG. 5 represents a first variant of the first embodiment of the invention. The same elements are indicated by the same reference signs as in FIG. 3. The system of FIG. 5 further comprises a bubble detector 82 disposed on the branch 13 between the autoinjector 80 and the input-output 52 of the first valve 50 and preferably near the inlet-outlet 52. The autoinjector 80 injects on the branch 13 of the fluid circuit a sample 30 interposed between a first air bubble 31 and a second air bubble 32. When the detector 82 detects the first bubble 31, which arrives at the detector 82 before the sample 30, the detector 82 triggers a discharge signal of the first bubble 31. A control system makes switch the first valve 50 following the signal evacuation. According to one aspect of the invention, the switching time of the first valve 50 is determined by the quotient between, on the one hand, the volume of the fluid circuit comprised between the detector 82 and the inlet-outlet 52 of the valve 50 and the other hand the flow rate of the fluid in the fluid circuit. Those skilled in the art will use a commercially available bubble detector, for example of the ultrasonic detector type or an optical detector. When the detector 82 detects the second bubble 32 located after the sample, the detector 82 triggers a detection signal of a second bubble. This signal can be used to trigger either the evacuation of the second bubble 32, in a similar manner to the first bubble, or a change of direction of circulation during the period of the back and forth.
    [0024] FIG. 6 represents a first variant of the second embodiment of the invention. The same elements are indicated by the same reference signs as in Figure 4A-4B. The system of FIG. 6 further comprises a bubble detector 82 disposed on the branch 13 between the autoinjector 80 and the input-output 73 of the valve 70 and preferably close to the input-output 73.
    [0025] The autoinjector 80 injects on the branch 13 of the fluid circuit a sample 30 interposed between a first air bubble 31 and a second air bubble 32. When the detector 82 detects the first bubble 31, which arrives at the detector 82 before the sample 30, the detector 82 triggers a discharge signal of the first bubble 31. A control system switches the valve 70 from the first state to the second state, following the evacuation signal of the first bubble 31. Advantageously, the switching time of the valve 70 is determined by the quotient between, on the one hand, the volume of the fluid circuit comprised between the detector 82 and the inlet-outlet 73 of the valve 70 and, on the other hand, the flow rate of circulation. fluid in the fluid circuit. When the detector 82 detects the second bubble 32 located after the sample, the detector 82 triggers a detection signal of a second bubble. This signal can be used to trigger either the evacuation of the second bubble 32, similar to the evacuation of the first bubble, or a change of direction of circulation for the duration of the back and forth. FIG. 7 illustrates a second variant of a system for transferring a fluid sample according to the first embodiment of the invention. The same elements are indicated by the same reference signs as in Figure 3A-3C. The system of FIG. 7 further comprises two additional valves 26, 27 disposed respectively on bifurcations 24, 25 of the branch 23 connected to the inlet-outlet 62 of the second valve 60. The additional valves 26, 27 allow selectively orient the analyzed sample either to a container 94, for example a recovery tube, or to another container 95, which serves for example waste collection or trash. In a state where it is desired to eliminate the sample analyzed, the valve 26 is closed and the valve 27 is open, so as to orient the fluid sample from the inlet-outlet 62 of the valve 60 to the branch 25, via the valve 27 and thus to the trash container 95. In the state where it is desired to recover the sample analyzed, the valve 26 is open and the valve 27 closed, so as to orient the fluid sample of the input-output 62 of the valve 60 to the branch 24, via the valve 26 and thus to the container 94, for example a recovery tube. FIG. 8 represents a second variant of a system for transferring a fluid sample according to the second embodiment of the invention. The same elements are indicated by the same reference signs as in Figure 4A-4B. The system of FIG. 8 further comprises two additional valves 26, 27 respectively disposed on bifurcations 24, 25 of the branch 23 connected to the inlet-outlet 71 of the valve 70. The additional valves 26, 27 make it possible to orient selectively the sample analyzed either to a container 94, for example a recovery tube, or to another container 95, which serves for example trash. In a state where it is desired to eliminate the sample analyzed, the valve 26 is closed and the valve 27 is open, so as to orient the fluid sample from the inlet-outlet 74 of the valve 70 to the branch 25, via the valve 27 and thus to the trash container 95. In the state where it is desired to recover the sample analyzed, the valve 26 is open and the valve 27 closed, so as to orient the fluid sample of the input-output 74 of the valve 70 to the branch 24, via the valve 26 and thus to the container 94, for example a recovery tube. FIG. 9 schematically represents a third variant of a transfer system of a fluid sample according to the first embodiment of the invention, which combines the first and the second variant. The same elements are indicated by the same reference signs as in FIGS. 3A-3C, 5 and 7. The operation of the various elements is similar to that described with reference to FIGS. 3A-3C, 5 and 7. FIG. schematically a third variant of a transfer system of a fluid sample according to the second embodiment of the invention, which combines the first and the second variant. The same elements are indicated by the same reference signs as in FIGS. 4A-4B, 6 and 8. The operation of the various elements is similar to that described with reference to FIGS. 4A-4B, 6 and 8. FIG. schematically a complete system for transferring a fluid sample according to one embodiment of the invention. An autoinjector 80 comprises a source microplate 82 having wells, each well containing a sample to be analyzed. A needle 83 can be moved in front of each of the wells of the microplate 82. A two-state injection valve 84, connected to a loop 85, and a pump 86 make it possible to take the samples in the microplate 82. Another pump 87 allows the fluids to be continuously circulated in the fluidic detection circuit. The fluidic detection circuit comprises the fluidic cell 10, and two valves 28, 29 which make it possible to select either a buffer solution reservoir 98 or a trash container 81. The autoinjector 80 has two main states depending on the position of the valve 84, which is either in the position of loading a sample in the fluidic circuit (as illustrated in FIG. 11), or in the injection position of the sample towards the fluidic cell 10 (not shown). In the loading state, pump 86 sucks via needle 83 along the path from needle 83, valve 84, loop 85, valve 84, to vessel 81. First, pump 86 draws a bubble from defined air volume when the needle 83 is positioned outside the microplate 82, then a predefined volume of sample when the needle 83 is positioned in a well of the microplate 82 and, finally, again an air bubble . Meanwhile, in the loading state, the pump 87 continuously draws a buffer solution from the tank 98, the valve 28 being open and the valve 29 closed. The pump 87 circulates the buffer solution to the fluidic cell 10. In the injection state, during the allers of the injection, the pump 87 can suck the circulation buffer fluid from the reservoir 98, the valve 28 being open and the valve 29 closed. The pump 87 pushes the buffer fluid through the loop 85 which contains the sample framed by the bubbles, to the fluid cell 10 along the path 98, 28, 87, 84, 85, 84, branch 13, valve 50, branch 11 , input-output 1. The sample thus arrives progressively towards the fluidic cell 10. During the returns of the injection, the pump 87 can suck the fluids coming from the fluidic cell 10 and push them towards the trash container 81, the valve 28 being closed and valve 29 open, according to branch path 23, valve 60, branch 12, inlet-outlet 2, fluidic cell 10, inlet-outlet 1, branch 11, valve 50, branch 13, valve 84, loop 85 , valve 84 towards the trash container 81. The sample is gradually backwards. The alternation of these back and forth during the injection state makes it possible to go back and forth. During this time, the pump 86 is stopped. The system and method of the invention can be implemented on existing fluid analysis systems. The adaptation of an analysis system involves only minor modifications.
    权利要求:
    Claims (5)
    [0001]
    REVENDICATIONS1. A fluidic sample transfer system (30) in a fluidic cell (10), the fluidic system comprising: a fluidic cell (10) having a first input-output (1) disposed upstream of said fluidic cell (10) and a second input-output (2) disposed downstream of said fluidic cell (10), a first fluid circuit (11, 12, 13) connected to said first input-output (1) and a second fluid circuit (21, 22, 23) connected to said second input-output (2), first injection means (80) connected to the first fluid circuit (11, 12, 13), said first injection means (80) being adapted to inject in series in the first fluid circuit (11, 12, 13): a buffer solution (35), a first volume of separation fluid (31) followed by a fluid sample (30) and then a second volume of separation fluid ( 32) and another buffer solution (36), circulation means adapted to circulate said ec fluid sample (30) interposed between the first volume of separating fluid (31) and the second volume of separating fluid (32) in the first fluid circuit (11, 12, 13) towards the first input-output (1) ) of the fluidic cell (10), characterized in that the system comprises: first discharge means (41, 43, 50, 70) disposed on the first fluid circuit (11, 12, 13) near the first an inlet-outlet (1), said first discharge means (41, 43, 50, 70) being adapted to extract, upstream of the fluidic cell (10), said first volume of separation fluid (31) and, respectively , said second volume of separating fluid (32), second injection means (42, 44, 60, 70, 90, 91) disposed on the second fluid circuit (21, 22, 23) near the second inlet output (2), said second injection means (42, 44, 60, 70, 90, 91) being adapted to inject, downstream of the cell; a fluid volume (10), a third volume of separation fluid (33) between said buffer solution (35) and the fluid sample (30) and, respectively, a fourth volume of separation fluid (34) between the fluid sample (30) and said other buffer solution (36), and bi-directional flow means adapted to circulate the fluid sample (30) in a first direction of flow from said first input-output (1) to said second input- output (2) or in a second direction of flow from said second input-output (2) to said first input-output (1) without passing the first, second, third or fourth volume of separating fluid (31, 32, 33, 34) in said fluid cell (10).
    [0002]
    A fluidic fluid sample transfer system in a fluid cell according to claim 1, wherein the first discharge means (41, 43, 50, 70) comprises a first valve (50) having at least a first port an input-output (51) fluidly connected to the first input-output (1) of the fluidic cell (10), a second input-output port (52) fluidly connected to the first injection means (80), and a third input-output port (53), said first valve (50) having at least one first state, wherein the first input-output port (51) of the first valve (50) is connected to the second port input-output device (52) of the first valve (50) and a second state, wherein the second input-output port (52) of the first valve (50) is connected to the third input-output port (53) of the first valve (50).
    [0003]
    The fluidic fluid sample transfer system in a fluidic cell according to claim 2, wherein the second separation fluid injection means (42, 44, 60, 70) comprises a second valve (60) having at least a first input-output port (61) fluidly connected to the second input-output (2) of the fluid cell, a second input-output port (62), and a third input-output port (63); ) fluidly connected to a source of separation fluid, said second valve (60) having at least one first state, wherein the first input-output port (61) of the second valve (60) is connected to the second port of an input-output (62) of the second valve (60) and a second state, wherein the second input-output port (62) of the second valve (60) is connected to the third input-output port ( 63) of the second valve (60).
    [0004]
    The fluidic fluid sample transfer system in a fluidic cell according to claim 1, wherein the first separation fluid discharge means and the second separation fluid injection means comprise a valve (70) having at least one minus a first input-output port (71) fluidly connected to the first input-output (1) of the fluidic cell (10), a second input-output port (72) fluidly connected to the second input-output (2) of the fluidic cell (10), a third input-output port (73) connected to the first injection means (80), a fourth input-output port (76) fluidly connected to a source of separation fluid (90), a fifth input-output port (75) and a sixth input-output port (74), said fifth and sixth input-output ports (75, 74) being preferably connected fluidically to an evacuation or recovery device (81, 93, 94, 95), said valve ( 70) having at least one first state in which the first input-output port (71) is connected to the third input-output port (73) and wherein the second input-output port (72) is connected at the sixth input-output port (74), said valve (70) having at least one second state in which the third input-output port (73) is connected to the fifth input-output port (75) and or wherein the fourth input-output port (76) is connected to the sixth input-output port (74).
    [0005]
    Fluidic fluidic fluid transfer system in a fluidic cell according to one of claims 1 to 4, comprising a detector (82) disposed on the first fluid circuit (13) between the first injection means (80). and the first means for discharging the separation fluid (41, 43, 50, 70), the detector (82) being adapted to detect the first volume of separating fluid (31) and / or the second volume of separating fluid (32). A system for transferring a fluidic sample into a fluidic cell according to one of claims 1 to 5, wherein the third volume of separating fluid (33) being a volume of air, the second fluid injection means of separation (42, 44, 60, 70) comprises an air pump (91) preferably connected to an air filter (90). System for transferring a fluidic sample into a fluidic cell according to one of claims 1 to 6, wherein the fluidic cell (10) has a shape ratio greater than the aspect ratio of the first and second fluid circuits (11). , 12, 13, 21, 22, 23). System for transferring a fluidic sample into a fluidic cell according to one of claims 1 to 7, wherein the fluidic cell (10) comprises a biochip adapted for analyzing the fluid sample in at least one site of analysis, and preferably in a matrix of analysis sites, the analysis being preferably an optical analysis, for example of the surface plasmon resonance (SPR) resonance type, with localized surface plasmon resonance (LSPR), with Resonant mirrors, Bloch surface wave (BSW), integrated waveguides 30 or resonant microcavities (WGM). 9. A system for transferring a fluidic sample into a fluidic cell according to one of claims 1 to 8, comprising synchronization means adapted to synchronize the operation of the first means for discharging the separation fluid (41, 43, 50, 70), second separating fluid injection means (42, 44, 60, 70, 90, 91) and bidirectional circulation means. 10. A fluidic sample transfer system in a fluidic cell according to one of claims 1 to 9, wherein the fluidic system comprises en6. 7. 8.25a reservoir (93, 94, 95) adapted to receive the fluid or fluids from the second input-output (2) of the fluidic cell (10). 11. A method for transferring a fluid sample in a fluidic system, the method comprising the following steps: series injection into a first fluid circuit (11, 12, 13) of a buffer solution (35) followed by a first a volume of separating fluid (31), a fluid sample (30), and a second volume of separating fluid (32) and another buffer solution (36), circulation of the fluid sample (30). ) interposed between the first separation fluid volume (31) and the second separation fluid volume (32) in the first fluid circuit (11, 12, 13) towards the first input-output (1) of the cell fluidic medium (10), evacuation, near the first inlet-outlet (1), of the first volume of separating fluid (31), upstream of the fluidic cell (10), passage of the buffer solution (35) followed by the fluid sample (30) in the fluid cell (10); injecting, near the second inlet-outlet (2), a third volume of separation fluid (33) between said buffer solution (35) and the fluid sample (30) downstream of the fluidic cell (10) and bi-directionally circulating the fluid sample (30) interposed between the second separation fluid volume (32) and the third separation fluid volume (33) in a first flow direction from said first input output (1) to said second input-output (2) and / or in a second direction of flow from said second input-output (2) to said first input-output (1) without passing the second or third volume of separating fluid (32, 33) in said fluid cell (10); evacuation, near the first inlet-outlet (1), of the second volume of separating fluid (32), upstream of the fluidic cell (10), passage of the fluid sample (30) in the fluid cell (10) ) followed by the other buffer solution (36) through the fluid cell (10) of the first input-output (1) to the second input-output (2); injecting, near the second input-output (2), a fourth volume of separation fluid (34) between the fluid sample (30) and the other buffer solution (36) downstream of the fluidic cell ( 10).
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    同族专利:
    公开号 | 公开日
    CN106471375B|2018-11-13|
    US20170067805A1|2017-03-09|
    WO2015124873A1|2015-08-27|
    FR3017956B1|2018-02-09|
    US10302533B2|2019-05-28|
    EP3108260A1|2016-12-28|
    CN106471375A|2017-03-01|
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    法律状态:
    2015-12-03| PLFP| Fee payment|Year of fee payment: 3 |
    2016-12-01| PLFP| Fee payment|Year of fee payment: 4 |
    2017-12-01| PLFP| Fee payment|Year of fee payment: 5 |
    2019-12-02| PLFP| Fee payment|Year of fee payment: 7 |
    2021-11-12| ST| Notification of lapse|Effective date: 20211005 |
    优先权:
    申请号 | 申请日 | 专利标题
    FR1451411|2014-02-21|
    FR1451411A|FR3017956B1|2014-02-21|2014-02-21|SYSTEM AND METHOD FOR TRANSFERRING A FLUIDIC SAMPLE IN A FLUIDIC CELL|FR1451411A| FR3017956B1|2014-02-21|2014-02-21|SYSTEM AND METHOD FOR TRANSFERRING A FLUIDIC SAMPLE IN A FLUIDIC CELL|
    EP15709281.8A| EP3108260A1|2014-02-21|2015-02-19|System and method for transferring a fluid sample in a fluid cell|
    PCT/FR2015/050406| WO2015124873A1|2014-02-21|2015-02-19|System and method for transferring a fluid sample in a fluid cell|
    CN201580019743.6A| CN106471375B|2014-02-21|2015-02-19|System and method for the transfering fluid sample in fluid compartment|
    US15/120,358| US10302533B2|2014-02-21|2015-02-19|Fluid sample analysis system and method for transferring a fluid sample in a fluid cell using two-way circulation|
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