EP4255629A1

EP4255629A1 - Pcr detection chip, associated test kit and analysis system for implementation

Info

Publication number: EP4255629A1
Application number: EP21811104.5A
Authority: EP
Inventors: Maël LE BERRE; Claire LEBOUTEILLER; Bilal HADJI
Original assignee: Bforcure
Current assignee: Bforcure
Priority date: 2020-12-04
Filing date: 2021-11-29
Publication date: 2023-10-11
Also published as: US20240033730A1; FR3117045A1; WO2022117493A1; CN116528981A

Abstract

A system for testing a microfluidic sample chip for testing biological samples, notably for analysis of the PCR and/or fluorescence type, having the form of a hollow block comprising at least one chamber delimited by an upper wall and a lower wall into which chamber can be introduced a sample that is to be tested and which is associated with thermalization means and with fluorescence measurement means. According to the invention, the system further comprises a tab, preferably opaque and/or rigid or semirigid, positioned in the continuation of, and preferably in the same plane as, the walls.

Description

DESCRIPTION

TITLE: PCR detection chip, associated test device and implementation analysis system

[0001] The invention relates to a microfluidic sample chip for testing biological samples, in particular for a PCR and/or fluorescence type analysis, associated test apparatus as well as a biological sample analysis system using such a chip.

To perform a rapid real-time PCR (Polymerase Chain Reaction) reaction, it is necessary to modify the temperature of the sample mixed with its reagent as quickly as possible while preferably measuring its fluorescence (here -after the rapid "cycling" of the sample). A method and a device of this type are described for example in patent application WO201 1/138748.

The PCR reaction is generally implemented in a disposable container because at the end of the reaction, the large-scale amplification of the DNA target to be detected contaminates the surface of the container with the target to be amplified, which prevents its reuse. The PCR reaction containers are therefore so-called consumable containers.

[0004] These PCR consumables must be able to be handled and inserted into the sample test device in a simple, obvious and rapid manner to facilitate their use, in particular in a context where operational efficiency is important.

[0005] A specific implementation of PCR is real-time PCR where DNA amplification is measured during the reaction by a fluorescence signal from a probe whose fluorescence depends on the progress of the amplification reaction. In this case, an important issue of rapid cycling technologies is the design of a consumable receiving the PCR reagent which allows good thermal transmission to the sample so that the temperature of the sample quickly equilibrates with the temperature of the sample. the thermal cycling device (or thermal cycler) while ensuring optical accessibility to the sample. [0006] WO 2018/1 14625 describes in particular a microfluidic sample chip containing a chamber, preferably flat and/or thin, formed between an at least partially transparent face and a heat-conducting face, for example made of aluminium, the transparent side allowing the PCR reaction to be monitored using an optical means, for example a fluorescence probe, while allowing a very rapid variation in the temperature of the sample thanks to its heat-conducting side. This chip can be used for example in combination with a microfluidic temperature control system as described in this application WO2018/1 14625, making it possible to very quickly modify the temperature of the sample between its two extreme values typical of PCR technology , in a time of the order of a second.

[0007] The object of the present invention is to make the structure and the use of the chip described in WO 2018/1 14625 even simpler, more reliable and more efficient so as in particular to facilitate its industrialization, and in particular to meet even better to current needs for rapid diagnostic orientation tests or to respond in emergency contexts which require the ability to perform reactions such as PCR in a few minutes.

The invention relates to a system for testing a microfluidic sample chip for the PCR-type test of biological samples, this chip having the shape of a preferably parallelepiped block comprising at least one hollow chamber for receiving a sample, delimited in particular by a first wall (or lower wall) made of a material with high thermal conductivity, preferably metallic, and a second wall (or upper wall) made at least partially of a transparent material, this block having at least two openings allowing the introduction of the sample into at least one of the chambers and the evacuation of the atmosphere present in the chamber during the introduction of the sample, the first and second walls being parallel to each other, the chip further comprising a tab, preferably at least partially opaque and/or rigid or semi-rigid, arranged in the extension and preferably in the same plane as the walls s, the chip also comprising a sealing device, arranged in contact with the second wall or upper wall, characterized in that it comprises a side wall provided with an opening allowing the passage of the chip towards thermalization means, complementary foolproofing means intended to cooperate with foolproofing means present on the chip, means for holding the chip in contact with the thermalization means after introduction of the latter through the opening which maintain the chip in contact with the thermalization means, contact preferably generated by the cooperation of foolproofing means located on the chip and complementary foolproofing means, in order in particular to start the sample analysis sequence, this system comprising further means for measuring the fluorescence of the sample.

[0009] According to a preferred mode, the chip test system according to the invention will include fluorescence imaging means.

[0010] According to one embodiment, the test system according to the invention has the shape of a rectangular lamella preferably comprising near one of its ends an opening surrounded by a frame preferably one of which at least of the sides is opaque, in particular the side of the frame which is on the side of the tongue, opening in which is housed on one side a plate of material with high conductivity, preferably metal, forming the first wall and the on the other side the means for holding the chip forming the second wall, the plate of high conductivity material, the block of transparent material and the frame forming the active part of the chip.

[0011] According to a preferred embodiment, the system according to the invention will include keying means consisting of a notch located on the chip, preferably located on the tab.

According to one embodiment, the chamber has grooves to facilitate the flow of the sample in the chamber.

According to a preferred embodiment, the system according to the invention will comprise an adhesive sealing film located on the side of the block secured to the tongue, extending in the direction of the opening without being secured to the block, the latter comprising at least two openings communicating with the chamber, said sealing film being designed to adhere to said block and seal the openings after filling the chamber with the sample.

According to one embodiment, the system according to the invention will comprise a protective film which can be removed before use of the chip, this film being placed on the openings of the transparent block so as to avoid any contamination of the front chamber introduction of the sample.

[0015]According to an alternative, the system according to the invention will comprise an identification label at the tongue. [0016] In a preferred variant, the second wall has a groove arranged at its periphery on the side of the chamber in which an adhesive is arranged to firmly secure the second wall to the first wall.

In a variant of the invention, the system according to the invention will comprise at least four chambers arranged side by side and forming a square. In this case, it will include an opaque wall placed between each of the rooms to reduce light transmission between the different rooms and with the outside environment.

[0018] Preferably, the tab will be at least partially opaque in order to prevent the passage of external light to the sample once the chip has been inserted into the associated test device. In addition, the tab will preferably be rigid or semi-rigid, arranged in the extension and preferably in the same plane as the walls.

According to a preferred embodiment, the chip according to the invention comprises a sealing means integral with the chip, arranged in contact with the upper wall in order to confine the sample inside the chamber or chambers once it inserted into the chip, which makes it possible to avoid any contamination of the environment by leakage of the sample.

[0020] The sealing device integral with the chip will preferably be a sealing film. The sealing film will preferably be placed at least on each of the openings so as to avoid any contamination from the outside by the sample to be analyzed and therefore to avoid distorting the following results. This sealing film can be prepositioned on the plate during the manufacturing stage (see below). This sealing film makes it possible to seal the openings when the chambers have been filled with their sample to be analyzed.

[0021] The sealing film will preferably be self-adhesive located on the side of the block, integral with the tongue, extending in the direction of the opening without being integral with the block, the latter comprising at least two openings communicating with the chamber, said sealing film being designed to adhere to said block and seal the openings after filling the chamber with the sample.

[0022]According to a variant, the chip will also comprise a protective film. The protective film can be removed before use of the chip, this film being placed on the openings of the transparent block so as to avoid any contamination of the chamber, in particular between the manufacture and the transport of the chip, an external contamination being able to distort the result of the analysis.

[0023] Thus the protective film will be in contact with the upper wall of the chip and will be removed during the filling of the chamber or chambers. As soon as the filling is finished, the sealing film is immediately positioned over the openings, in a sealed manner, in order in particular to allow a rise in pressure in the chambers, as explained below.

[0024] In this way, the use of sterile packaging of the chip after its manufacture until its use is avoided (as is usually the case for this type of device)

[0025]According to another variant, it is the protective cover of the sealing film which acts as a protective barrier between the potentially contaminating exterior and the interior of the chamber before the chip is used.

According to a preferred form, the chip according to the invention has the shape of a rectangular lamella preferably comprising near one of its ends an opening surrounded by a frame preferably of which at least one of the sides is opaque, in particular the side of the tongue allowing its insertion into the associated test device, opening in which is housed on one side a plate of high conductivity material, preferably metal, forming the first wall and the other side the block forming the second wall, the plate of high conductivity material, the block (preferably made of transparent material) and the frame forming the active part of the chip. When the frame is opaque (or partially opaque at least on the tab side) this prevents external light from entering the chamber and causing noise in the fluorescence measurement signal. This takes on even more importance in the case of multi-chamber chips, where the fluorescence from each chamber must be independently detectable. The opaque walls then make it possible to "optically isolate" the rooms from each other and from the outside environment. Thus the tab may have an opaque portion adjacent to the active part of the chip, of sufficient length to be visible outside the test device after introduction of the chip into the PCR analysis device.

The sample chip can comprise at least 2 chambers, that is to say more generally n chambers (with n being an integer greater than or equal to 2). In one case particular, the n=i*j (with i and j two integers greater than or equal to 1) rooms are arranged side by side and form a square (if i=j) or a rectangle (if i different from j).

According to a preferred variant, the sample chip according to the invention comprises keying means intended to cooperate with complementary keying means located in the test device to analyze the sample contained in the chamber in order in particular to start the Sample analysis sequence automatically when the chip is inserted into the PCR analyzer and to identify if the chip has been inserted upside down.

These keying means consist for example of a notch located on the chip, preferably located on the tab.

[0030] Preferably, the rectangular slat has a notch on one of its sides having a keying function.

[0031] The chip may also include an identification label at the tab.

In order to facilitate the flow of the sample in the chamber, the latter will preferably include grooves.

To seal the second wall tightly to the first wall, the sample chip according to the invention will preferably include a groove arranged at its periphery on the side of the chamber in which an adhesive will be deposited.

According to a variant embodiment, the sample chip according to the invention will comprise four chambers arranged side by side and forming a square.

According to a preferred embodiment, the sample chip according to the invention is a rectangular wafer of length equal to at least twice, preferably at least three times, the width of this wafer in which there is a rectangular opening (or square ) near one of its ends, thus defining a frame around the opening. In this opening are housed on one side, (for example above), a window in transparent material (in a transparent material compatible with the PCR such as for example polypropylene, polymers and copolymers of cycloolefins (COC, COP ) and in general amorphous polymers, in transparent material, resistant to a temperature of at least 110C), with a flat outer surface reaching the level of the surface of the wafer and the inner surface of which comprises a hollow part forming the chamber to receive the sample to be analyzed. On the other side of the opening (for example below) is arranged an aluminum lamella (or any other material compatible with PCR and having a suitable thermal conductivity-see below) whose thickness will preferably be comprised between 100 and 500 microns, said slat resting on the outer edges of the window made of transparent material, the window and/or the slat being provided with grooves, preferably at their periphery, making it possible to seal the window and the metal lamella, for example using a bonding means such as a glue compatible with PCR (of the silicone type or equivalent). Preferably the metal strip will (slightly) protrude from the (lower) surface of the wafer to improve the thermal contact with the heating means of the test apparatus.

[0036] The rectangular wafer thus has a portion in the extension (preferably) of the frame, acting as a tongue for handling the chip (its active part consisting in particular of the window and the metal plate assembled using glue) and to insert and/or remove it from the test device in a simple way through a slot of complementary size to the cross section of the rectangular wafer and position it in a reproducible way in the test device. To facilitate the positioning of the wafer carrying the active part of the sample chip in the test device, provision will preferably be made for foolproofing means, preferably on the wafer, for example a notch on one of the sides of the strip, these coding means being associated with complementary means placed in the test device so as to indicate to the user that the wafer is correctly positioned in the device when the coding means engage in the complementary means (it is possible to associate sound or light means for example indicating this engagement to the user who can thus trigger the analysis of the sample (either manually or automatically).

These keying means can also be used to automatically detect whether the chip has been inserted upside down in the device and warn the user by means of a light or sound signal or by means of the man-machine interface.

The rectangular plate (which includes both the frame and the tongue) will preferably be at least partially at the level of the frame, made of an opaque material (as defined below), for example an opaque black plastic material. compatible with PCR. It has in fact been observed that this opacity, in particular at the level of the frame and in particular when the chip has several chambers, it was possible to obtain a better reading of the fluorescence by allowing better insulation of the light between the various chambers and better insulation of the external light.

Of course, in order to be able to handle this wafer easily, the material(s) used must give it sufficient rigidity (rigid or semi-rigid material - see definition below) to allow its introduction into the slot. provided for this purpose in the test device.

[0040] The transparent material window will have at least two openings making it possible to reach the chamber into which the sample to be analyzed is introduced (preferably at each end of the chamber). Preferably, the chamber will include grooves to facilitate the flow of the liquid sample to be analyzed and to avoid the formation of bubbles so as to promote, during the filling of the chamber, the evacuation of the gaseous atmosphere contained therein. .

When the filling of the chamber is finished, the openings are sealed, for example using a transparent film compatible with the PCR and with the associated fluorescence measurement, a film which preferably covers the whole of the window. . This sealing film is preferably pre-positioned on the wafer during the manufacture of the chip according to the invention.

The tab according to another aspect of the invention can receive on one (or more) of its faces a QR Code or a bar code (or any other means of identification) making it possible to identify the chip. It is also possible, given the space available on the tab, to provide a label on which the user can note information.

[0043] In order to protect the chip chamber from any contamination, in particular between manufacture and use of the chip (transport), provision is made to cover the openings with a protective film that does not leave any residue liable to inhibit the PCR reaction.

If the tongue is preferably flat and arranged in the same plane as the walls, a variant of the invention consists in providing a tongue having for example the shape of a protuberance, preferably arranged in the extension of the walls, allowing a better grip of the chip to introduce it into the test device. [0045] Preferably, an opaque wall will be placed between each of the chambers (for a chip with multiple chambers) making it possible to reduce the light transmission between the various chambers and with the external environment.

The sample test device according to the invention may, according to a variant, comprise a side wall having an opening allowing the passage of the chip towards thermalization means, complementary keying means intended to cooperate with the keying means present on the chip, a block of glass which can move vertically in the direction of the chip after introduction of the latter through the opening and come to press the chip on the thermalization means on receipt of a command generated by the cooperation of keying means and complementary keying means.

[0047] Preferably, the test device will include means for measuring fluorescence and in particular means for imaging by fluorescence arranged so as to allow the passage of light from and to the imaging means.

In this patent application, the following terms will have the following meaning:

[0049] The term "PCR compatible material" means a material which does not contain PCR inhibitors, does not degrade enzymes, fluorescent probes, dNTPs, oligo nuclides and does not absorb oligonucleotides and other RNA or DNA sequences and does not release molecules that could interact with the functioning of enzymes or change the characteristics of the PCR reaction.

[0050] The term "opaque material" means a material whose optical transmission in the visible range in particular, i.e. between 300 nm and 800 nm, is less than 20%, preferably less than 1%, for a thickness of material one millimeter.

[0051] The term "transparent material" means a material whose optical transmission in the visible range, in particular, is greater than 80%.

[0052] The term “high thermal conductivity material” means a material whose thermal conductivity is greater than 15 wm-1 .K-1 . The term "rigid or semi-rigid material" means a material whose Young's modulus is greater than 10 MPa.

The terms device, test device, sample test device, PCR analysis device are used interchangeably to designate the PCR sample analysis device.

The invention will be better understood using the following examples of embodiment, given without limitation, together with the figures which represent:

[0056] [Fig. 1] Figure 1, a top view of the sample chip according to the invention.

[0057] [Fig. 2] Figure 2, a bottom view of the sample chip from Figure 1.

[0058] [Fig. 3] Figure 3, a sectional view of the active part of the chip along the axis A-A of Figure 1.

[0059] [Fig. 4] Figure 4, a sectional view of the active part of the chip along the B-B axis of Figure 1.

[0060] [Fig. 5] Figure 5, a schematic view of the chip in combination with the test device.

[0061] [Fig. 6] Figure 6 an exploded view of the chip according to the invention before bonding of the elements of the active part of the chip.

[0062] [Fig. 7] Figure 7, the different steps of bonding the window and the metal part to make the active part of the chip.

[0063] [Fig. 8] Figure 8 is a schematic top and bottom view of a multi-chamber sample chip, without the metal pad.

[0064] [Fig. 9] Figure 9, a sectional view of the multi-chamber chip of Figure 8 along the C-C axis in this figure.

[0065] [Fig. 10] FIG. 10, curves representing the fluorescence signal of different samples (RFU on the ordinate) as a function of the number of cycles (on the abscissa). [0066] [Fig.1 1] Figure 11, the various stages of removal and bonding of the protective and sealing films.

In these figures the same elements bear the same references.

In Figure 1 the sample chip according to the invention is shown according to a preferred embodiment in the form of a rectangular wafer 10. This parallelepiped-shaped wafer 10 made of opaque material has a length approximately five times its width on the figure (preferably between 50x10mm (and a thickness of at least 1mm) and 200x100mm (and a thickness of at most 10mm)). It is provided in its active part 13, (left part of the figure) with a substantially square opening 16 (see FIG. 6) surrounded by an opaque frame 1 in which is housed a transparent window 2. This is provided with two openings 5 located on a diagonal of the window 2 and communicating with the chamber 4 (see FIGS. 3 and 4) provided with grooves 12, said openings 5 allowing one of them to introduce the sample to be analyzed and for the other the evacuation of the atmosphere present in the chamber 4. In the extension and in the same plane as the frame 1 is the tongue 9 provided on one of its sides with a notch 8 serving as foolproofing means during the introduction of the rectangular plate 10 in the test device, not shown in the figure. In this figure 1 is shown a top view of the sample chip, while figure 2 shows a bottom view of the same sample chip. In this figure 2, a metal plate 3 is housed in the opening 16, thus closing the active part 13 of the sample chip. This metal plate 3, in the extension of the tongue 9, will be slightly raised (above) relative to the tongue 9 in order to achieve good thermal and mechanical contact with the thermalization element 21 (figure 5).

[0069] Figure 3 shows a sectional view along the axis AA (diagonal of the active part 13) defined by the two openings 5 (or injection holes). The transparent window 2 rests using its peripheral end 17 on a flange 18 integral with the frame 1, leaving a groove 7 into which an adhesive (silicone or other) will be injected all around the window 2 making it possible to fix sealingly the metal plate 3 to the window 2 and to the frame 1 . The metal plate 3 thus delimits the chamber 4 into which the sample is introduced through the injection holes 5. A sealing film 6 covers the openings 5 either as a seal to seal the openings 5 after injection of the sample to be analyzed in chamber 4. FIG. 4 represents a view in section along the axis BB of the active part of the sample chip. In the chamber 4 are represented several grooves 12 which facilitate the circulation of the liquid sample as well as its distribution in the chamber.

[0071] Figure 6 shows an exploded view of the sample chip of Figure 1, before the assembly of the window 2 and the metal plate 3 inside the opening 16 in the rectangular wafer 10.

[0072] Figure 7 illustrates the assembly of the window 2 and the metal plate 3. Using an injection means 19 of semi-liquid adhesive, we just fill the groove 7 arranged around the window 2 with the semi-liquid adhesive 14 such as a silicone or other material compatible with PCR (FIG. 7b) then the metal plate 3 (FIG. 7c) is applied to the window 2 so as to seal the chamber 4 thus created. Then the adhesive is polymerized (hardened) using a suitable UV 15 source. The sample chip being thus prepared, it is possible to introduce the sample to be analyzed into the chip, then seal the openings 5 with the aid of an adhesive PCR film and proceed with the analysis of the sample as shown in the figure. figure 5.

For this we have a test device, left part of Figure 5 (inside). This comprises holding means, for example, a mobile transparent block 20 (preferably made of glass) through which the fluorescence measurement can be made and a sample thermalization element; (for more details on a thermalization element, reference may be made to application WC201 8/114625 in the name of the Applicant). On the other side of the wall 23 of the test device (on the right in FIG. 5: exterior) is introduced through the opening 22 in this wall, the rectangular plate 10 thanks to the tongue 9, so as to bring the active part 13 of the chip above and into contact: the adequate positioning of the chip on the thermalization element 21 is achieved thanks to the keying notch 8 cooperating with complementary keying means located in the testing device. This cooperation then triggers a movement (downwards in FIG. 5a) of the glass block 20 which presses the active part 13 of the sample chip (FIG. 5b) ensuring good thermal contact of the chip (whose metal part is at the contact of the thermalization element) which makes it possible to start the thermal cycling of the sample. During this cycling, the optical signal 25 resulting from the fluorescence of the sample is collected on the sensor 24, allowing real-time monitoring of the fluorescence in the chip. After a predetermined period (approximately 40 cycles) the thermal cycling is stopped and reassemble the glass block 20 (figure 5c) in its rest position, the chip being pulled outwards thanks to the tongue 9.

The chip is handled from outside the device and its insertion does not require the device to be opened. No access panels, drawers or doors are required on the test device, hence its simplicity and speed of use.

The chip is inserted through a fixed slot, then removed in the same way once the detection operation is complete.

The correct positioning of the chip is determined by the notch on the edge of the chip. Once in position, it engages a contactor (not shown in Figure 5) which allows the locking of the system.

[0077] The holding and thermalization means form in this variant a mobile jaw which makes it possible to hold the chip in position, it is composed of: a glass block mounted on a motorized platform allowing its vertical movement for the upper part, a thermalization element (metallic for example), allowing rapid temperature change during PCR thermal cycles for the lower part.

The pressure applied to the chip is preferably between 100g and 10kg (ideally distributed evenly over the entire surface of the chamber),

[0079] Once the PCR is complete, the jaw opens automatically (under the control of an automaton programmed for this purpose, in a manner known per se) releasing the chip, which can be removed by the user (FIG. 5c) .

The locked position under pressure (FIG. 5b) makes it possible to ensure maximum contact between the metal plate of the chip and the thermalization element, on the one hand, and to prevent the boiling of the sample liquid contained in the chip when the temperature is around 100°C on the other hand. But it also makes it possible to avoid any leakage of PCR products into the device which could contaminate it.

The transparent block, preferably made of glass, makes it possible to collect the fluorescence signal from the sample during the PCR amplification. [0082] FIG. 8 represents a variant embodiment of the invention in which the chip according to the invention comprises 4 chambers, each being provided on one side with a metal surface for the rapid transfer of heat via the thermalizing element and on the other a transparent window to allow the excitation and the optical detection of the fluorophores. Each chamber can receive a different sample to be analyzed.

For the one-chamber variant or the four-chamber variant, a protective film is positioned on the window, covering the injection holes, in order to avoid contamination of the chamber during transport of the chip. This film is removed and discarded just before injecting the sample into the chip.

In Figure 1 1, the sealing film 6 sticker, cut to size, is prepositioned on the tab of the chip. Only part of the sealing film is stuck on the tab, the other part, covering the window, remains protected by the protective cap until the sample is injected. Thus, the user can easily remove the protective cap and stick the sealing film to obstruct the injection holes after the insertion of the sample in the chamber, without having to worry about positioning the film correctly or cut.

More precisely, FIG. 11 describes the variant of the configuration with two films: the sealing film 6 and the protective film 27. The protective film 27, making it possible to avoid contamination of the chamber before its use , during transport for example, is removed before inserting the sample into the chip, fig 11a. The protective cover 26 of the sealing film 6 is removed at the time of sealing the chip, after injection of the sample and, in FIG. 11b, the sealing film 6 is folded over the upper wall of the chip in order to hermetically cover the openings used to fill the chip, fig 1 1 c.

[0086] A label presenting a QR code, for example, makes it possible to automatically reference each of the chips, which makes it possible to have patient monitoring and a connection with the Laboratory Information Management System (LIMS) of the hospitals for medical applications. The label can also present information concerning the volume and the number of chambers available on the chip.

Figure 9 is a sectional view of the chip of Figure 8 along the (diagonal) axis CC. As before the grooves 7 must be filled with glue to fix and seal the assembly. FIG. 10 represents the RFU fluorescence signal as a function of the number of cycles to which the samples are subjected. The TARGET 1 and TARGET 2 curves correspond to two DNA targets of the SARS-COV-2 coronavirus while the CTRL curve corresponds to the internal control. The amplification is clearly detectable from the ^35th cycle, which corresponds to rapid detection in less than 20 minutes.

Such a structure of the chip with its sealing film, its tab and its keying system makes it possible in particular to introduce the chip into the test device through the slot 22 without having to open the device.

Claims

CLAIMS System for testing a microfluidic sample chip for the PCR type test of biological samples, this chip having the shape of a block (10) preferably parallelepiped comprising at least one hollow chamber (4) for receiving a sample, delimited in particular by a first wall (3) (or bottom wall) made of a material with high thermal conductivity, preferably metallic, and a second wall (2) (or top wall) made at least partially of a transparent material, this block (10) having at least two openings (5) allowing the introduction of the sample into at least one of the chambers (4) and the evacuation of the atmosphere present in the chamber (4) during introduction of the sample, the first and second walls (3, 2) being parallel to each other, the chip further comprising a tongue (9) preferably at least partially opaque and/or rigid or semi-rigid, arranged in the extension and preferably in s the same plane as the walls (2,3), the chip also comprising a sealing device, disposed in contact with the second wall or upper wall (2), characterized in that it comprises a side wall (23) provided an opening (22) allowing the passage of the chip towards thermalization means (21), complementary foolproofing means intended to cooperate with foolproofing means (8) present on the chip, holding means (20) of the chip in contact with the thermalization means after introduction of the latter through the opening (22) which hold the chip in contact with the thermalization means (21) contact generated by the cooperation of keying means (8) located on the chip and additional foolproofing means, in order in particular to start the sample analysis sequence, this system further comprising means for measuring the fluorescence of the sample. Chip test system according to the preceding claim, characterized in that it includes fluorescence imaging means. Test system according to one of the preceding claims, characterized in that the chip has the shape of a rectangular lamella (10) preferably comprising near one of its ends an opening (16) surrounded by a (1) preferably of which at least one of the sides is opaque, in particular the side of the frame which is on the side of the tongue, opening in which is housed on one side a plate of material with high conductivity (3), preferably made of metal, forming the first wall (3) and on the other side the means for holding the chip forming the second wall (2), the plate of high conductivity material (3), the block of transparent material (2) and the frame (1) forming the active part of the chip. System according to one of the preceding claims, characterized in that the foolproofing means (8) consist of a notch (8) located on the chip, preferably located on the tab (9). System according to one of the preceding claims, characterized in that the chamber (4) has grooves (12) to facilitate the flow of the sample in the chamber (4). System according to one of the preceding claims, characterized in that it comprises an adhesive sealing film (6) located on the side of the block (10) integral with the tab (9), extending in the direction of the opening (16 ) without being integral with the block, the latter comprising at least two openings (5) communicating with the chamber (4), said sealing film being designed to adhere to said block (2) and close the openings (5) in a sealed manner after filling the chamber (4) with the sample. System according to one of the preceding claims, characterized in that it comprises a protective film which can be removed before use of the chip, this film being placed on the openings (5) of the transparent block so as to avoid any contamination of the chamber (4) before introduction of the sample. System according to one of the preceding claims, characterized in that it comprises an identification label at the tongue. System according to one of the preceding claims, characterized in that the second wall (2) comprises a groove (7) arranged at its periphery on the side of the chamber (4) in which an adhesive (14) is arranged to securely fasten seals the second wall (2) to the first wall (3). System according to one of the preceding claims, characterized in that it comprises at least four chambers (4) arranged side by side and forming a square.

11. System according to claim 10, characterized in that an opaque wall is arranged between each of the chambers making it possible to reduce the light transmission between the various chambers and with the external environment.