WO2015075396A1 - Sample test system and method - Google Patents

Abstract

A sample test system and method. ABSTRACT: A sample test method that comprises providing a first device (10) comprising first and second opposing faces (11 and 12), and comprising a network of wells (20) extending between the first face (11) and the second face (12) of the first device (10), each well (20) opening on at least the first face (11) and each well (20) containing a reagent, and that comprises providing at least one sample (40), and that comprises bringing the first face (11) of the first device (10) into contact with the sample (40), with at least two wells (20) each sealingly facing at least one respective portion of the sample (40), so as to form an assembly (80), then bringing the reagent into contact with the sample (40). FIGURE 7

Description

 System and method for sample testing.

The present invention relates to systems and methods for testing a sample.

In the field of histology, and especially high-throughput screening, for the testing of biological samples such as tissues, high throughput is achieved with the Tissue Micro Array (TMA) System. It allows to evaluate by immunohistochemistry the expression of the same protein on hundreds of samples of the same tissue of different individuals, or to evaluate the expression of the same protein on different tissue samples. of the same animal (heart, spleen, liver, lungs, ...). Such a system remains limited to the search for the expression of a target protein on a large number of samples arranged on the same slide or on the same support. The present invention is intended to overcome these disadvantages by allowing to perform a large number of different analyzes on the same support, for example simultaneously.

For this purpose, the invention relates to a sample testing method comprising:

Providing a first device having opposite first and second faces, and comprising an array of wells extending between the first face and the second face of the first device, each well opening on at least the first face and each well containing a reagent, - Provide at least one sample,

Put the first face of the first device into contact with the sample, with at least two wells facing each with at least a respective portion of the sample in a sealed manner, to form an assembly,

Put the reagent in contact with the sample.

Thanks to these provisions, it will be possible to test several different antibodies on the same slide. More generally, a large number of biological samples can be tested simultaneously and by different reagents, aiming at an increase in the flow of analyzes performed. In the same perspective of high-speed test method, the same biological sample can be tested by several reagents simultaneously, allowing statistical studies. Such a test method goes in particular in the direction of a possible automation of the system.

As another advantage it will also be possible to optimize, quickly on the same medium, experimental protocols.

In preferred embodiments of the biological sample test method according to the invention, one or more of the following provisions may also be used:

 - the sample is a biological sample.

 the reagent is brought into contact with the samples by agitating the assembly.

the at least one sample is carried by a second device comprising a first and a second opposite face and bearing samples on at least one of its faces, and for which the first face of the first device and one of the faces of the second device are contacted with at least one well facing at least a portion of at least one sample carried by at least one of the faces of the second device.

 at least one of the wells opens on the two opposite faces of the first device.

 the method comprises the following steps:

 Position the assembly so that the first device is above the second device,

Fill the at least one of the wells of the first device with the second face of the first device. the method comprises observing by optical means the samples from the second face of the first device.

 the method comprises closing the outlet of the at least one well on the second face of the first device by a plate.

In preferred embodiments of the biological sample test system according to the invention, one or more of the following may also be used:

The system comprises at least a first device comprising opposite first and second faces, and comprising a network of wells extending between the first face and the second face of the first device, each well opening on at least the first face and each well. containing a reagent, the system also comprising at least one sample, and the system being adapted to contact the first face of the first device with the sample, with at least two wells facing each with at least a respective portion of the sample sealingly, to form an assembly adapted to contact the reagent and the sample.

The system further comprises at least one second device comprising a first and a second opposite face adapted to carry samples on at least one of its faces and adapted so that the first face of the first device and one of the faces of the second device are brought into contact with at least one well of the first face of the first device facing at least a portion of at least one sample carried by at least one of the faces of the second device.

the first face of the first device is locally hydrophobic around the outlet of at least one well on the first face of the first device. - The system further comprises a reservoir adapted to be connected to at least one well of the first device, said reservoir being adapted to individually fill said well.

- The system further comprises bores previously machined in the first device, said bores being adapted to assist the separation of the first device and the second device after contacting the reagent and the sample.

the first device is made of polymer and is suitable for thermo-molding wells.

- The device is anti-ultraviolet material at least on the surface.

at least one of the wells has a conical geometry and for which the outlet section of said well on the first face (11) is greater than the section of the well between the first face and the second face of the first device.

Other features and advantages of the invention will become apparent from the following description of several of its embodiments, given by way of non-limiting example, with reference to the accompanying drawings.

 On the drawings:

 FIG. 1 shows the system according to one embodiment of the invention with the first device,

 FIG. 2 shows a conical well geometry,

 FIG. 3 schematizes the individualized filling of the wells,

 FIG. 4 shows the common filling of all the wells by a common tank,

 FIG. 5 schematizes the steps of the test method for the system according to the embodiment of the invention with the first device,

 FIGS. 6a-6d illustrate possible sealing configurations of the first device,

 FIG. 7 shows the system according to one embodiment of the invention with the first and the second device,

 FIGS. 8a and 8b show the maintenance of the first and second devices by a hybridization chamber,

 FIGS. 9a-9d schematize the steps of the test method for the system according to the embodiment of the invention with the first and the second device,

 FIG. 10 shows the bores in the first device,

Figure 11 shows the first device with With open wells on both sides of the part, FIGS. 12a-12d diagrammatically illustrate steps of the test method in the case of wells opening on both sides of the part.

 FIG. 13 illustrates the case of wells emerging on each face and of conical geometry,

 FIG. 14 illustrates the case of wells supplied with solution or gas by lateral connectors,

 Figure 15 illustrates the automation of the system.

In the different figures, the same references designate identical or similar elements. In all that follows, we note z a vertical axis in space as shown in Figure 1. We also note x and y two axes perpendicular to each other and perpendicular to z so that the axes (x, y , z) form an orthonormal frame.

The system according to a first embodiment comprises a first device 10 comprising opposite first and second faces, 11 and 12 as shown in FIG.

The first device 10 is made of biocompatible material. In a variant, it is made of a non- ^bio¬ compatible material whose inner surface of the wells is ^bio¬ compatible. It is easy to clean and reuse frequently. It may be for example a glass plate with a thickness of about one centimeter, for example. The first device 10 may also be a plate of polymer material adapted or subjected to a suitable surface treatment. Indeed polymer materials can generating an aspecific binding of the biological molecules (proteins, nucleic acids, etc.) and a preliminary surface treatment with a hydrophobic compound will ensure reliability in the concentration of the compounds to be tested.

The first device 10 comprises a network of wells 20 which can be made extending between the first face 11 and the second face 12 of the first device 10. A well network 20 may comprise, for example, wells 20 arranged in an ordered manner in lines. and in columns. The lines then define the x direction. Each well 20 opens on at least the first face 11. The wells 20 may for example be cylindrical with an axis parallel to z. A cylindrical well 20 may have any fixed section as a function of z as shown in FIG. 1. In another embodiment, the geometry of the wells may be adapted according to the intended applications. The section of the well 20 may for example be variable as a function of z. The well 20 may also for example have a conical shape as shown in FIG. 2. A well 20 will have, for example, a diameter size of between 1 mm and 15 mm, preferably between 5 mm and 10 mm, depending on the intended applications. .

The wells 20 will typically have a depth of the order of a few millimeters, and which may be correlated to the diameter of the wells.

In a glass plate, the wells 20 are produced for example by ultrasonic machining through a mask or by etching or by hot pressing. against a preform. An attack by hydrofluoric acid HF can be used to improve the final surface state of the wells. In the case of machining wells 20 in a polymeric material, the shaping can be done by heat treatment, by molding for example.

Each well 20 is adapted to receive a reagent. The wells 20 open on the first face 11, and are closed on the opposite side. They can receive a surface treatment beforehand which also determines the size of the wells according to the desired treatment.

The density and size of the wells 20 of the first device 10 are parameters that may vary depending on the size of the materials and the volume of reagent that must be present in the wells 20. This reagent may be different from a well 20 to the same. other. Some wells 20 of the first device 10 may also for example be left empty as shown in FIG.

The first device 10 may be translucent to allow continuous or sequential temporal observation of the reagent contained in the wells 20 by optical or spectroscopic means. The reagent may be for example a liquid, a solid optionally in the form of a powder, for example an aerosol.

The filling of the wells 20 can be done simultaneously for all the wells 20 by contacting the first face 11 of the first device 10 on which the wells 20 open, with a reservoir containing the reagent for filling the wells 20. The filling by a common tank, as shown in FIG. 4, for example, aims at saving time, and ultimately, for example, automating the process.

The wells 20 can also be individually filled with identical reagents in each well 20 or with different reagents.

The system of the invention also comprises for example at least one sample 40 of dimensions ranging from a few microns (μm) to a few centimeters (cm). It may be for example a biological sample: skin, tissue or cells. It may be a piece of polymer for industrial applications, or a plant sample, any material or an electromechanical micro or nano-structure. The structure may be for example passive function such as a film with a substrate or active function such as a sensor.

In the embodiment which will be described here, it will be considered for the sake of simplification the case of a biological sample.

The first device 10 is brought into contact with the biological sample 40 as shown in FIG. 5. For this, the biological sample 40 is positioned on the first device 10 so that, if the biological sample 40 is to be tested preferably on one side, it is this face of the biological sample 40 which faces the first device 10. At least one well 20 is placed facing at least a portion of the biological sample 40. The biological sample 40 is of sufficient spatial extent to cover several wells adjacent to each other, and of such rigidity that, at rest, it does not return to contact with the reagent contained in the wells. The reagent and the biological sample are then brought into contact. For this, the assembly 80 formed by the biological sample 40 and the first device 10 held together is for example returned around an axis of the plane (x, y) so that, under the effect of gravity, the reagent contained in the at least one well 20 comes into contact with the biological sample 40. The assembly 80 is then returned again around an axis of the (x, y) plane, towards its initial position for example, so as to the reagent contained in the well 20, s'1 is liquid, down by gravity bottom wells 20. The sample 40 and the first device 10 are separated. To bring the reagent into contact with the biological sample, the assembly 80 may also alternatively be turned around an axis of the plane (x, y) more than twice or shaken vertically or more generally agitated, before separating the sample 40 and the first device 10. In all that will follow the term "agitation" will be used generally to describe both a simple reversal of the system around an axis of the plane (x, y), as several consecutive reversals of the system around an axis of the plane (x, y), a translational movement of the system in any direction, a periodic movement of the system, a random movement in direction and periodicity suitable for the brought into contact with the reagent and the biological sample. Such "agitation" may in particular be automated.

The tightness between the wells 20 of the first device 10 must be ensured to avoid cross-well contamination during agitation, for example of the assembly 80, for example if different liquids are placed in each well 20. The first face 11 of the first device 10 on which the wells 20 open will for example undergo a surface treatment to be rendered locally hydrophobic between the wells 20 The first face 11 will be treated by mechanical texturing, by chemical treatment or by deposition of material on the first face 11 of the first device 10. The first face 11 will for example be treated by a deposition of silica nanoparticles.

The first face 11 of the first device 10 will for example be subdivided into small zones surrounding each of the wells, and separated from each other by a hydrophobic zone so that after contacting and stirring the assembly 80, the reagent such as a liquid, contained in the wells 20 can not pass between the wells 20, that is to say that the junction between the wells 20 is sealed as shown in Figure 6a.

The hydrophobicity can be obtained by texturing the surface or by adding a hydrophobic material or whose surface structure makes it hydrophobic.

The first face 11 may for example be rendered hydrophobic over its entire surface, and then locally treated to release certain areas of hydrophobicity, by treatment through a mask for example, such as etching through a mask . These areas of the first face 11 made hydrophobic may be lines delimiting the wells 20. These lines may be thick (Figure 6b) or multiple such as double lines (Figure 6c). The first face 11 may alternatively be locally treated to be rendered hydrophobic only in certain areas, by treatment through a mask directly for example.

 The seal between the wells 20 can also be provided by silicone grease spread around said wells 20. For example, a pasty product, such as a grease, is used which is placed around the well, for example in the form of 'a rope. A non-flowing product is preferably chosen, so that the latter is not likely to be placed in the well, and / or has a high chemical neutrality (for example with respect to antibodies) so as not to influence the test. Silicone grease has shown superior sealing performance over other tested products in comparative tests using two glass plates. Alternatively the first face 11 is coated with grease which enters the wells 20. These are then cleaned to remove the grease that has entered. The tightness between the wells 20 can also be ensured by spacers 70 placed between the wells 20, as shown in FIG. 6d, tightly assembled to the first device 10, and sealed to the sample 40.

Sealing could also be ensured by using an adhesive polymer or metal film which would be adhered to the device 10, thereby sealing the wells on the surface 12 side.

In this embodiment, an initially frozen tissue disk to be tested is for example selected, and brought into contact with the reactive liquid placed in the wells 20 of the first device 10, for example 1 at 2 mm two by two. In order for the response of the tissue to the reactive liquid to be representative, a minimum surface of the fabric will have to be brought into contact with the test liquid via one or more wells. The characteristic dimension of the smallest surface will, for example, typically be order of 600pm.

As an alternative to this embodiment, it may be for example a method for performing allergy tests, made on the skin of the patient that will constitute the biological sample 40. The skin is initially made sensitive by needle-sticking or scraping. Wells 20 are filled with different reagents between wells 20 which are allergenic substances. The wells 20 are then brought into sealed contact with the surface of the skin undergoing the test. The patient's arm may for example be positioned above the first device 10, then the entire arm and the device 10 is assembled by tape for example. The patient then shakes his arm for example and then returns to the initial position. The patient may also for example turn his arm to contact the allergens and his skin for an hour or more generally shake his arm attached to the device 10. After the patient has returned his arm to its initial position, the first device 10 is separated from the patient's arm on which the reagents reacted with the patient's skin. The method described above thus makes it possible, for example, to carry out the allergology test.

In this embodiment the same fabric will be sectorized to perform a series of different analyzes. The individualized filling of the wells will make it possible to test different reactive compounds or different concentrations of reagent during the same test. In another example of this embodiment, in agronomy one can for example test the impact of different plant protection products or different doses on plant leaves or the impact of different pathogens on the same leaf. This type of test can be done on plants in culture. In this example, the device operates in an uncontrolled environment, in the sense that, unlike laboratory tests, it is necessary to integrate the effects of solar radiation, temperature variations, rain, etc. This is therefore the case in such cases. conditions, to ensure that the products tested on the plants do not degrade and that the test remains valid.

To avoid the effects of solar radiation in particular, it is possible to envisage several solutions that are to use a material constituting the device that is not transparent to UV, or to carry out an anti-UV surface treatment (non-transparent to UV) on the device 10. or add another material, such as fabric or veil for example, around the area on which the tests are performed.

In such an embodiment, for testing outside a laboratory, the device may for example be instrumented, typically with a temperature probe or a "patch" affected by UV radiation or if its temperature exceeds a certain value . This condition could be shown in any detectable way, such as a change of color for example. Alternatively for other applications, for example industrial applications, for which larger quantities of products have to be tested over large areas, much larger device and well dimensions could be envisaged.

For all of the above embodiments, the reaction can be monitored, for example, in an instrumented manner, in situ using spectroscopic methods.

According to a second embodiment, the system of the invention may also comprise a second device 30, which has first and second opposite faces 31 and 32 as shown in FIG.

This may be for example a glass plate 30 such as a microscope plate. The glass plate 30 can typically have dimensions between 1cm and 10cm, for example 2.5cm by 7cm.

The second device 30 is adapted to receive at least one biological sample 40 on at least one of its faces. It may be for example the face 31 receiving a biological sample 40. The biological sample 40 may be for example a tissue sample. In this embodiment, the biological sample 40 is secured to the face 31 of the second device 30 using any suitable mechanical and / or chemical means for accessing a portion of the biological sample. For example, paraffin is used which allows the positioning and adhesion of the biological sample 40 to the face 31 of the second device 30. The second device 30 is then placed on the first device 10 so that the face 31 of the second device 30 is facing the first face 11 of the first device 10 as illustrated in FIG. Figure 9a. In order for the test method to be implemented, the biological sample 40 is positioned on the second device 30 so that at least one well containing reagent is contacted with at least a portion of the biological sample. 40. The assembly 80 formed by the first device 10 and the second device 30 carrying the biological sample held together is then agitated so that the reagent contained in at least one of the wells 20 is brought into contact with the biological sample 40. The first device 10 and the second device 30 are held together by clips, tape or any other suitable means.

A pressure means may be used to ensure a good seal at the interface between the first device 10 and the second device 30 for the necessary time. The inter-well seal can be provided by mechanical pressing, for example between the first device 10 and the second device 30. In this case, particularly in the case of a low roughness of the microscope slide, the adhesion between the first device 10 and the second device 30 may be strong.

Bores previously machined in the first device 10 and in correspondence with others in the second device 30 may for example be adapted to ensure that the first device 10 and the second device 30 are brought into sealed contact by using a rod, for example threaded and through the two devices 10 and 30 and keeping them in contact via bolts positioned on either side of the assembly 80 and which maintain the contact. The tightness may also for example be ensured by the use of two other surfaces sandwiching the assembly 80 to improve the contact between the first device 10 and the second device 30.

Alternatively, the inter-well seal is provided in any of the ways described above for the first embodiment.

In a variant, the first device 10 and the second device 30 are held together using a hybridization chamber 100. The hybridization chamber comprises a frame 102, for example a metal frame, a positioning support 110, a positioning slide 109.

Specifically, the assembly 80 of the first device 10 and the second device 30 is held by the frame 102 of the hybridization chamber 100 which traps the edge of the assembly 80 as shown in Figure 8a. The frame 102 comprises an upper part 108, a lower part 106 separated and a connecting element 112 providing the connection between the upper part 108 and the lower part 106.

The assembly 80 is positioned on the positioning slide 109. The positioning slide 109 is for example a frame allowing the first and second devices 10, 30 to be held together and relative to one another. In particular, positioning makes it possible to geometrically align the wells 20 of the first device 10 with the Samples of the second device 30. The positioning drawer 109, on which the assembly 80 has been loaded, is placed on the positioning support 110. The positioning drawer 109 is alternately bonded to the positioning support 110. The positioning support 110 110, thus loaded with the positioning slide 109 itself loaded with the assembly 80, is then moved, for example slid between the lower portion 106 and the upper portion 108 of the hybridization chamber 100.

The lower portion 106 is of variable thickness, which increases continuously between the end opposite the connecting member 112 and the end adjacent the connecting member 112, as shown in Figure 8b.

Thus, during the horizontal sliding of the loaded positioning support 110, between the lower part 106 and the upper part 108 of the hybridization chamber 100, the assembly 80 mounted on the positioning slide 109 will be kept compressed between the lower part 106 and the upper portion 108 of the hybridization chamber 100. A screw 104 is used to translate the loaded positioning support 110 into the interior of the hybridization chamber 100. The screw 104 also makes it possible to block the slide positioning device 109 on which is loaded the assembly 80 in the hybridization chamber 100, as shown in Figure 8b.

The screw 104, inserted into a hole horizontally traversing the connecting element 112 and opening on either side, will be screwed inside a threaded hole 114 pierced horizontally in the setting drawer 110. The screw 104 is accessible for clamping, for example with a key type BTR or other. Actuation of the screw 104 thus causes the slide to slide into a position where it is held tight by the beveled shapes of the lower part 106 and the positioning support 110. To release the assembly 80 and its setting drawer position 110, it will loosen ie turn the said screw in the opposite direction.

The first device 10 can be inserted before the upper part 108 of the hybridization chamber 100 and the second device 30 carried by the positioning slide 109 can be inserted between the upper part 108 and the lower part 106 of the chamber hybridization 100.

Alternatively, the upper and lower portions 106, 108, and the connecting member 112, will be molded together.

 Thus, according to one aspect, independently of other considerations, if one seeks to keep the assembly 80 sealed, an invention relates to a hybridization chamber 100 comprising at least one frame 102, a positioning support 110, a drawer of positioning 109, a screw 104,

the frame 102 comprising an upper portion 108, a lower portion 106, and a connecting member 112 between the upper portion 108 and the lower portion 106, the lower portion 106 having an increasing thickness between an end opposite the connecting member 112 and an end adjacent to the connecting element 112,

the positioning slide 109 being adapted to receive an experimental device, the positioning support 110 being adapted to receive the positioning slide 109, the positioning support 110 comprising a threaded hole 114,

the screw 104 being adapted, by its insertion into the threaded hole 114, to drive and hold the positioning support 110 between the upper part 108 and the lower part 106.

 The variation of thickness of the lower part makes it possible to ensure a good maintenance of the experimental device mounted on the positioning slide.

 The clamping of the positioning support 110 between the two upper portions 108 and lower 109 by a screw system located in the thickness of the positioning support makes it possible to disengage the experimental device.

According to another aspect, the characteristic described here, when compatible with this invention:

 The upper part 108 is pierced with a gap.

Such a gap allows access from the outside of the hybridization chamber 100 to the experimental device placed in the hybridization chamber. Then the assembly 80 is agitated. The first device 10 and the second device 30 are then separated as shown respectively in FIGS. 9a to 9d. In this embodiment, the separation of the first device 10 and the second device 30 may be assisted for example by bores 90. The bores 90 may for example be through the first device 10 as shown in FIG. bear against the bores 90 on the second device 30, and apply to it a force allowing the detachment of the second device 30 of the first device 10.

In this configuration of the system, the biological sample 40 may be tested for example by several different reagents placed in the different wells 20 of the first device 10 all facing a respective part of the biological sample 40 during agitation of the assembly 80 and second device 30. When the first device 10 and the second device 30 are separated, a reagent has reacted with the biological sample 40 in each zone in which a reagent has been brought into contact with a sample zone biological 40.

In this embodiment, the second device 30 can receive several biological samples 40 to be tested, for example simultaneously. The second device 30 comprises for example a network of locations capable of receiving the biological samples 40, this network being corresponding to the well network 20 provided by the first device 10. The biological samples 40 are therefore, in this case, positioned to so that each biological sample 40 faces at least one well 20 of the first device 10. Some wells 20 may not face any biological sample 40.

The biological samples 40 are secured to the second device 30 which acts as a sample holder as described above, and for example by the use of paraffin. Each biological sample 40 is for example cast in paraffin and then placed on the second device 30.

The wells 20 of the first device 10 may contain an identical or different reagent between the wells 20.

The assembly 80 and second device 30 is then agitated to bring the biological samples 40 into contact with the reagents contained in the wells.

The biological samples 40 can all be tested by the same reagent or by different reagents contained in the different wells 20 compared with biological samples 40.

In one embodiment of the system in this configuration, samples of 0.6mm to 2mm in diameter and a few microns thick are taken from impregnated and paraffin-embedded tissues. These samples are then positioned on a plate acting as a second device 30. The wells 20 are then for example greater in diameter than the size of the biological sample 40 deposited on the plate. The diameter of the wells may be for example between 1mm and 15mm. The distance between the wells to avoid contamination may for example be 1mm. Alternatively we can have wells of 1mm in diameter, with a distance between two wells typically 2mm. The plate used to deposit the biological samples 40 may typically have dimensions of the order of 25x70cm, and a thickness of the order of 1 to 2mm.

In another embodiment, a culture The cell is deposited as a stopped cell culture on a microscope slide as a second device 30 for testing, for example, the effectiveness of the antibodies. The cell culture can be stopped by being fixed or frozen. The cell culture is fixed by a fixing agent, for example paraformaldehyde, acetone or ethanol.

In another embodiment, considering the absence of contamination problems, biological samples 40 can be placed on both faces 31 and 32 of the second device 30. In this embodiment, tests of each of the faces 31 and 32 can be made successively by contacting each of the faces 31 and 32 of the second device 30 with the reagent contained in the wells 20 of the first device 10 via contacting the first face 11 of the first device 10 successively with the faces 31 and 32 of the second device 30.

In another embodiment, at least one well 20 may also lead to the second face 12. In this embodiment, the bottom of the well 20 on the second face 12 may then consist of an additional glass plate 50, attached to the second face 12 of the first device 10 sealingly and removably as shown in Figure 11.

In this embodiment the device 10 'which can replace the first device 10 of the invention may consist of the first device 10 and a plate 50 for example of glass which is associated with the second face 12 of the first device 10 tightly to form the bottom. Such a configuration makes it possible, for example, easier to clean the wells 20 of the first device 10.

In this configuration, as shown in FIG. 12a, the first device 10 is positioned returned, that is to say with the second face 12 above the first face 11, and the first device 10 is positioned above the second device 30 having biological samples 40 assembled therewith. Wells 20 may be filled simultaneously or individually to the brim as shown in FIGS. 12a and 12b. A reservoir at the surface 12, on which each well opens, may for example allow filling of all the wells in a single operation as shown in Figures 12a to 12c. Alternatively the wells can also lead individually to the second face 12.

The biological samples 40 are therefore at this stage brought into contact with the reagents contained in the wells 20. A plate such as the additional glass plate 50 can then be positioned above the second face 12 of the first device 10 as shown on Figure 12c. The plate 50 may also make it possible to avoid evaporation of the reagent and the change of its concentration. The plate 50 can also for example help by adding the weight which seals the junction first device 10- second device 30. The contact between the first device 10 and the plate 50 is then sealed, before stirring for example all plate 50 first device 10 second device 30 if necessary alternatively. As an alternative to this configuration, the method may comprise only the first two steps illustrated in FIGS. 12a and 12b. The biological samples 40 brought into contact with the reagent contained in the wells can then be observed, for example, by optical means without having to separate the devices 10 and 30 and without having to empty the wells 20, the biological samples 40 having already been placed. in contact with the reagent (s). The observation is done for example through the reagent contained in the wells 20.

In the case of wells 20 opening on both faces 11 and 12, it is conceivable to manufacture the first device 10 by water jet machining.

Alternatively the first device 10 may also include at least one well 20 having a conical geometry. For example, in such a configuration, the outlet section of said well 20 on the first face 11 may be smaller than the section of the well 20 between the first face 11 and the second face 12 of the first device 10 as shown in FIG. such a configuration, in the case where the wells 20 open on either side of the first device 10, the first device 10 may be for example returned the second face 12 above the first face 11. The first face 11 of the first device 10 is brought into contact with the face 31 of the second device 30 comprising the biological samples 40, the outlets of the wells on the first face 11 being compared with the biological samples 40. The wells 20 can then be filled by the second face 12 of the first device 10. Each well 20 being filled by the face on which its section is the widest, towards the part where its section is the narrowest, the air trapped under the reagent during filling, in the case of a liquid reagent for example, can be more easily evacuated.

The biological samples 40 brought into contact with the reagent contained in the wells 20 can then be observed, for example, by optical means without having to separate the devices 10 and 30, the biological samples 40 having already been brought into contact with the reagent (s). .

The set 80 maintained, formed of the first device and the second device 30 carrying the biological sample, is then covered on the second face 12 of the first device 10 for example by a glass plate 50. The seal between the second face 12 the first device 10 and the plate 50 is then provided for example by mechanical pressing.

Alternatively, the system comprising the assembly 80 and the plate 50 held together, can then be turned over and the second device 30 comprising the biological samples 40 and located above can then be separated from the first device 10. This system can prior to being agitated.

In a particular embodiment, the reagent may also be gaseous. In this variant the filling of the wells 20 can be done for example by lateral connectors and transverse channels 61 as shown in Figure 14, at least one open face of the wells 20 of the device 10 being brought into contact with biological samples 40 of the device. in contact between the biological samples 40 and the wells 20 is directly provided by this configuration. The wells can also be supplied with solution by lateral connectors.

These side connectors, in the case of liquids or gases, can intercept wells at different heights, depending on their application. After the biological sample 40 is brought into contact with the reagent (s) and the biological sample 40 is separated from the device 10, the biological sample 40 is analyzed. In particular, the biological sample 40 is measured to determine the occurrence of a reaction between the biological sample 40 and the reagent (s). Such a measurement can be visual, optical or other.

Alternatively, in place of the second device 30 carrying the biological sample, a Tissue Micro Array (TMA) slide on which each sample can be analyzed independently. It will be possible, in this embodiment, to test several different antibodies and thus to validate, on the same slide, series of hybridomas during the synthesis of monoclonal antibodies (Ac) as well as on paraffin-embedded tissues. frozen tissues. It will also be possible to optimize, quickly on the same support, the protocols, for example for the use of an Ac immunohistochemistry (unmasking buffer, dilution of Ac, revelation kit).

In particular, the system can be used for immunoglobulin screening and for evaluation. simultaneous interaction of molecules with thin sections of tissue.

In the context of industrial applications, for example, the reaction of polymer pieces with chemical substances may be tested.

The automation of the system is for example applied to different steps of the method, as shown in Figure 15. The second device 30 is for example automatically conveyed from one end to the other of the process. The filling of the first device 10 is for example automatically in parallel with the process of conveying the second device 30 to lead to the assembly of the first and the second device. The assembly 80 is then shaken in a new automated step then the assembly is conveyed and disassembled. The first device 10 is transported to the cleaning station while the second device 30 is conveyed to the analysis station, provided for example with a microscope. The second device 30 is then conveyed after analysis to the output.

Claims

A sample test method comprising: Providing a first device (10) having opposite first and second faces (11 and 12), and comprising a well array (20) extending between the first face (11) and the second face (12) of the first device (10), each well (20) opening on at least the first face (11) and each well (20) containing a reagent, Provide at least one sample (40), Contacting the first face (11) of the first device (10) with the sample (40), with at least two wells (20) facing each with at least a respective portion of the sample (40) sealingly , to form a set (80), Contact the reagent with the sample (40). The method of testing samples according to claim 1 wherein the sample (40) is a biological sample. 3. A method of testing samples according to any one of claims 1 and 2 wherein the contacting of the reagent with the samples (40) is by agitation of the assembly (80). 4. A method of testing samples according to any one of claims 1 to 3 for which the at least one sample (40) is carried by a second device (30) having a first and a second opposite face (31 and 32). ) and carrying samples (40) on at least one of its faces, and for which the first face of the first device (10) and one of the faces of the second device (30) are brought into contact with at least one well (20) facing each other. with at least a portion of at least one sample (40) carried by at least one of the faces of the second device (30). The method of claim 4 wherein the assembly (80) is sealed using a hybridization chamber (100). 6. Method according to any one of claims 1 to 5 wherein at least one of the wells (20) opens on the two opposite faces (11, 12) of the first device (10). The method of claim 6 comprising the steps of: Position the assembly (80) so that the first device (10) is above the second device (30), Fill the at least one of the wells (20) of the first device (10) with the second face (12) of the first device (10). 8. A method according to any one of claims 6 and 7 comprising observing by optical means the samples (40) from the second face (12) of the first device (10). 9. A method according to any one of claims 6 to 8 comprising closing the outlet of the at least one well (20) on the second face (12) of the first device (10) by a plate (50). System comprising at least a first device (10) having opposite first and second faces (11 and 12), and comprising a well network (20) extending between the first face (11) and the second face ( 12) of the first device (10), each well (20) opening on at least the first face (11) and each well (20) containing a reagent, the system also comprising at least one sample (40), and the system being adapted to contact the first face (11) of the first device (10) with the sample (40), with at least two wells (20) facing each with at least a respective portion of the sample (40) in a sealed manner, to form an assembly (80) adapted to contact the reagent and the sample (40). 11. The system of claim 10 further comprising at least a second device (30) having a first and a second opposite faces (31 and 32), adapted to carry samples (40) on at least one of its faces and adapted for that the first face of the first device (10) and one of the faces of the second device (30) are brought into contact with at least one well (20) of the first face of the first device (10) facing at least a portion of at least one sample (40) carried by at least one of the faces of the second device (30). 12. The system of claim 11 further comprising a hybridization chamber (100), the assembly (80) being maintained sealed by said hybridization chamber (100). 13. System according to any one of claims 10 and 12 for which the first face (11) of the first device (10) is locally hydrophobic around the outlet of at least one well (20) on the first face (11) of the first device (10). 14. System according to any one of claims 10 to 13 comprising a reservoir (60) adapted to be connected to at least one well (20) of the first device (10), said reservoir (60) being adapted to individually fill said well (20). 15. System according to any one of claims 10 to 14 comprising bores (90) previously machined in the first device (10), said bores (90) being adapted to assist the separation of the first device (10) and the second device (30) after contacting the reagent with the sample (40). 16. System according to any one of claims 10 to 15 for which the first device (10) is made of polymer and suitable for thermo-molding wells (20). 17. System according to any one of claims 10 to 16 for which the device (10) is anti-ultraviolet material at least at the surface. 18. System according to any one of claims 10 to 17 for which at least one of the wells (20) has a conical geometry and for which the outlet section of said well (20) on the first face (11) is greater than the section of the well (20) between the first face (11) and the second face (12) of the first device (10).