Method of manufacturing biological analysis card comprising heating block

Abstract

The invention relates to a method for producing a biological analysis card (1), comprising: a step a) of providing a substrate (2) formed from:-at least two layers of films (3), one above the other,-a receiving space (4) formed between the two layers of films (3); a duct (5) leading into the receiving space (4); -a step b) of placing a plate (9) in the receiving space (4), said plate (9) comprising:-a first face (10a) and a second face connected by a contoured rim (11),-a plurality of holes (12) leading to at least the first face (10a) or the second face,-a plurality of channels (13) fluidly connecting the holes (12); a welding step c) of welding the two films (3) in the receiving space (4) to the plate (9) and welding the films (3) to sections of the edges (11) of the plate (9) by applying a heating block to the films (3) against the edges (11) on the sides of the first face (10a).

Description

Technical Field

[0001] The present invention relates to the field of biological sample analysis, and more specifically to a method for manufacturing an analysis card using a heating block, an analysis card obtained by the method, and a heating block used. Prior art

[0002] In vitro biological analysis of biological samples, such as polymerase chain reaction (PCR) tests, is based on one or more reactions between the biological sample and one or more reagents. The biological sample may include tissues and cells from the human or animal body and their derivatives, organs, blood, their components or derived products. There are microfluidic systems and methods for performing these biological sample analyses. The reagents are placed in the wells of a plate called an "array" and the biological sample to be analyzed is contacted with the reagents by flooding. The plate is assembled to a flexible base that includes a biological sample reservoir and conduits. The conduits in the base are connected to the channels in the plate in such a way that a fluid pathway is formed. An analytical card, such as a card for performing a PCR test, is formed. Analytical card. Assembling this card is a complex procedure that requires at least four layers of film and adhesive to be bonded to the board, which is then attached to the base. This involves the use of dedicated machines and a large number of consumables (film and adhesive layers), which significantly increases costs and increases the risk of defects when leakage occurs between layers.

[0003] In addition, the use of In the Assay Card configuration, only one biological sample is piped through the plate. It cannot be used to analyze several biological solutions, such as different concentrations of the same biological sample in the same analytical card, thus for example in the case of endotoxin detection tests. This is because in endotoxin detection tests, the biological sample is diluted to different concentrations and each dilution is analyzed. Therefore, great care must be taken to ensure that the different dilutions of the biological sample do not mix with each other before reaching the reaction wells of the plate, so that the reaction results are usable.

[0004] Finally, using the endotoxin detection test as an example, one specific aspect that must be pointed out is the thickness of the plate, which is The required detection sensitivity of endotoxins is usually greater than the reagent volume required for PCR testing, which affects the thickness of the reaction wells and thus the thickness of the analysis card, which must therefore be thicker (the intensity of the detected fluorescent signal is proportional to the thickness of the fluorescent liquid present in the reaction wells). This particular aspect poses a problem for sealing the plate in the base, since the surface of the plate is sealed between the membranes of the base (as in In the case of a PET-based system, the edges of the plate cannot be sealed, thus creating a risk of mixing biological samples or other liquids.

[0005] Therefore, there is currently no manufacturing method that allows for rapid, easy to produce and low-cost analytical cards with a thick plate between two base films, for example in the context of endotoxin detection tests.

[0006] Invention Introduction

[0007] The present invention therefore aims at producing an analytical card equipped with a plate of predetermined thickness capable of ensuring the separation of the fluid paths extending in the analytical card, for example making it possible to analyse several dilutions of the same biological sample or of different biological samples or of different biological samples with several dilutions in a more controlled way, without risk of mixing, and faster, easier and cheaper.

[0008] According to a first aspect, a method for manufacturing a bioanalysis card is provided, the bioanalysis card being used for performing a bioanalysis on a biological sample, the method comprising:

[0009] - Step a), providing a substrate consisting of:

[0010] - at least two overlapping films,

[0011] - a receiving space formed between the two films, delimited by a first boundary and an opening,

[0012] - a duct delimited by the contour and opening into the receiving space through a first boundary,

[0013] - Step b): Placing a plate in the receiving space by inserting it into the opening, said plate comprising:

[0014] - a first face and a second face connected by an edge having a profile,

[0015] - a plurality of holes opening into at least the first face or the second face,

[0016] - a channel with a plurality of fluid connection holes,

[0017] - a welding step c) consisting in welding the two films of the receiving space to the plate,

[0018] The method is characterized in that the welding step c) comprises welding the film to an edge section of the plate by applying a heating block to the film in close proximity to the edge of the first face side, the heating block comprising a support and a heating element, a first portion of the heating element protruding from the support at least during the application of the heating block, the first portion of the heating element then having a profile complementary to at least a section of the edge profile of the plate so that during the application of the heating block, when the film is pressed by the first portion of the heating element, the film takes the shape of the edge.

[0019] According to advantageous but non-limiting features, which may be achieved alone or in any combination:

[0020] - the profile of the edge segment of the plate is a straight line defining a convex angle between said segment of the edge of the plate and the second face, said convex angle being less than or equal to 90°, or a curve, the tangent of which defines a convex angle between said segment of the edge of the plate and the second face, said convex angle being less than or equal to 90°;

[0021] - the edge section to which the film is welded is a separating surface on the edge between the channels;

[0022] - a first portion of the heating element of the heating block comprises a non-heating zone, the width of which is greater than the width of the channel, so that when the first portion of the heating element is applied to the film next to the edge section, the non-heating zone faces the channel and the frangible valve;

[0023] - Each pipeline is equipped with at least one frangible valve located at one end of the pipeline;

[0024] - step c) comprises welding by means of a heating block of the film on the edge section of the plate to the borders between the pipes separating the pipes;

[0025] - step b) comprises the step of positioning the plate in the receiving space until the mouth of the channel is opposite to the mouth of the duct;

[0026] - step c) comprises the step of closing the opening of the receiving space by welding the two films to each other;

[0027] - the plate comprises a vacuum port connected to a channel connected to the hole, and the method comprises a vacuum step after the welding step c), the vacuum step consisting in piercing the film covering the vacuum port and blocking the channel connecting the vacuum port and the hole once the vacuum has been created;

[0028] - The thickness of the board is greater than 0.5 mm;

[0029] - welding the two films of the receiving space to the plate comprises welding the first film to the first face;

[0030] The heating block comprises a second portion of the heating element, the shape of which is complementary to the shape of the first face of the plate, and welding the two films of the receiving space to the plate comprises welding a first film to the first face, the first film being pressed against the first face.

[0031] According to a second aspect, an analytical card manufactured by the above method is proposed.

[0032] According to a third aspect, a heating block configured for implementing the above method is proposed, the heating block comprising a support and a heating element, a first portion of the heating element being configured to protrude at least during application of the heating block, the first portion of the heating element then having a profile complementary to at least a portion of the edge profile of the plate.

[0033] The heating block advantageously supplements the following features, which can be realized individually or in their various possible combinations:

[0034] - the first portion of the heating element comprises a non-heating zone, the width of which is greater than the width of the channel;

[0035] - the non-heating area of ​​the first part of the heating element is a groove;

[0036] - The first part of the heating element comprises a flexible nickel-chromium alloy (NiCr) element.

[0037] Detailed Description

[0038] Other features, objects and advantages of the present invention will become apparent from the following description, which is merely illustrative and non-limiting and must be read with reference to the accompanying drawings, in which:

[0039] Figure 1 is a diagram of the steps of a method of manufacturing an analytical card according to a possible embodiment of the present invention;

[0040] Figure 2 schematically shows an example of a base for an analytical card according to a possible embodiment of the invention;

[0041] Figure 3 shows an example of an analytical card during manufacture according to a possible embodiment of the invention,

[0042] Figure 4 schematically shows an example of an analysis card according to a possible embodiment of the invention;

[0043] Figure 5 is a perspective view of a portion of an analytical card according to a possible embodiment of the present invention;

[0044] Figure 6 Schematically showing a possible embodiment according to the invention, the heating block presses the first membrane against the edge of a plate of an analytical card having a contour forming a straight line;

[0045] Figure 7 Schematically showing a possible embodiment according to the invention, the heating block presses the first membrane against the edge of a plate of an analytical card having a curved profile;

[0046] Figure 8 An enlarged view of the assay card at the edge of the plate is shown;

[0047] Fig. 9 The heating blocks and plates are described.

[0048] The present invention relates to Figure 4 The manufacturing method of the analysis card 1 shown in FIG. Figure 1 The method is provided in detail, which first comprises step a): providing a base 2. Figure 2 The base 2 is formed by stacking two thermoplastic films 3. The material constituting the film 3 can be, for example, a composite based on polypropylene or polyethylene. The film 3 is a thin, soft, elastic surface. "Thin" means that the thickness of the film 3 is less than 200 μm. Preferably, the film 3 has a rectangular shape. More preferably, the two films 3a, 3b have the same shape when stacked. Therefore, it is impossible to distinguish Figure 2 The two films 3a and 3b in the figure are as follows, because one film is placed on top of the other film. Figure 5 Both films 3 are thermoplastic, which means that when heated to a certain temperature threshold, the films 3 soften, allowing them to undergo mechanical deformations that become fixed when the films 3 cool. The temperature threshold at which the films 3 soften and become deformable depends on the material constituting the films 3. The material constituting the films 3 is preferably a polymer, such as polypropylene or polyethylene, or a composite film based on polypropylene or polyethylene. Typically, the temperature threshold required to soften the films 3 is higher than 60°C.

[0049] The receiving space 4 is formed between the two films 3. More specifically, the receiving space 4 is comprised between two free film 3 portions. "Free" means that the films 3 are not welded to each other. In the plane of the base 2, the receiving space 4 is delimited by a first boundary 7 and an opening 8. The first boundary 7 is the area where the two films 3 are welded together. Figure 2 In the example shown, the first boundary 7 forms a straight line parallel to the width of the film 3 and along which the two films 3 are welded to each other. This welding can be performed, for example, using a laser that melts the film 3 along a precise path so that the two films 3 are welded to each other along this path. Other boundaries along which the two films 3 are welded together can define the receiving space 4.

[0050] The opening 8 corresponds to the area of ​​the membrane 3 which is not welded together and is free up to its edges. The opening 8 makes the receiving space 4 accessible from the outside. Preferably, the receiving space 4 has the same shape as the plate it is to receive, generally rectangular.

[0051] The opening 8 may include one or more of the three different edge portions of the film 3 excluding the first boundary 7 and may be located at Figure 2Thus, in this case, the opening 8 comprises an edge portion of the film 3 parallel to the first border 7 and two edge portions of the film 3 perpendicular to the first border 7. Each edge portion of the film 3 is free and not welded to an edge portion of another film 3. Figure 2 In the preferred embodiment shown, the first border 7 and the second border 230 of the base 2 form a pocket 23 so that the opening 8 is located only on one side of the membrane 3 .

[0052] The base 2 also includes a conduit 5 formed between the two films 3 and defined by a profile 6. The conduit 5 is similar to a channel formed between the two films 3, which allows the circulation of a fluid (e.g., a liquid biological sample). The profile 6 of the conduit 5 forms a curve along which the two films 3 are welded to each other in the same manner as the first boundary 7. Each conduit 5 is defined by two profiles 6. The profiles 6 of different conduits 5 are connected by a conduit-to-conduit boundary 7a, where the two films 3 are welded to each other. The conduit-to-conduit boundary 7a forms a part of the first boundary 7. The base 2 may also include a variety of elements, such as a reservoir formed between the films of the base 2 and connected to the conduits 5. Each conduit 5 may also include one or more frangible valves that block the conduits 5. For example, these frangible valves may be defined by an area (measuring a few millimeters or centimeters) where the film 3 is not welded firmly, and are configured to break according to the pressure applied to the frangible valve. The frangible valve is preferably located near the receiving space 4, in other words, at the end of the conduit 5 leading to the receiving space 4. For example, a reservoir in the form of blisters may contain the biological sample. The reservoir can be pressurized until the pressure applied to the frangible valve in the conduit 5 connected to the reservoir reaches a certain pressure threshold required to rupture the frangible valve. Once the frangible valve ruptures, the biological sample contained in the reservoir will flow in the conduit 5 as far as the receiving space 4.

[0053] The manufacturing method then comprises a step b) of arranging the plate 9 in the base 2 so as to finally obtain a base 2 equipped with the plate 9, such as Figure 3 As shown. The plate 9 is generally defined as an element having a flat surface and having a certain thickness, which is, however, very small compared to the dimensions of its flat surface. For example, the thickness is at least 10 times smaller than the width and length of the face. Preferably, the plate 9 has a thickness greater than or equal to 0.5 mm. In the example shown, the plate 9 has a rectangular surface, but other shapes may also be chosen.

[0054] Preferably, the plate 9 is formed of a thermoplastic material at least on its surface. For example, the plate 9 can be made of a material such as PP, PE, PMMA, PC, PS, POM, ABS, COP, etc. Figure 5As shown schematically, the plate 9 consists of two faces 10a, 10b opposite to each other and separated by a small thickness, i.e., the thickness is at least 10 times smaller than the width and length of the faces 10a, 10b. The maximum and minimum dimensions of the faces 10a, 10b of the plate 9 (in this case, the length and width of the faces 10a, 10b) are preferably greater than 2 cm, also preferably less than 10 cm, more preferably less than 4 cm. The thickness of the plate 9 is preferably less than 5 mm, more preferably less than 3 mm, and the thickness of the plate 9 is preferably greater than 0.5 mm. The faces 10a, 10b are connected by a side including an edge 11. In other words, the edge 11 corresponds to another face of the plate 9, which is different from the faces 10a and 10b and has a small width, i.e., at least 10 times smaller than the width and length of the faces 10a, 10b. Therefore, the length of the edge 11 is equal to the length or width of the faces 10a, 10b of the plate 9. Preferably, the profile of edge 11 is inclined relative to faces 10a, 10b and is therefore not perpendicular to faces 10a, 10b. Figure 6 , the profile of edge 11 is a straight line which defines a convex angle θ between edge 11 and second face 10b of plate 9 which is less than 90°. In other words, the smaller of the two angles defined by the profile of edge 11 between edge 11 and second face 10b (i.e. the angle less than 180°) is less than 90°. In other words, edge 11 preferably forms a chamfer. According to Figure 7 In another embodiment shown, the profile of the edge 11 is a curve, the tangent of which defines a convex angle θ between the edge 11 of the plate 9 and the second face 10b, which is less than 90°. Figure 7 In other words, the smaller of the two angles defined by the contour of edge 11 between edge 11 and second face 10b (i.e., the angle less than 180°) is less than 90°. According to this embodiment, the contour of edge 11 is preferably a convex curve.

[0055] The plate 9 includes a plurality of holes 12, which lead to at least one face 10a, 10b of the plate 9. The holes 12 may also pass through the plate 9 from one face 10a to the other face 10b. The holes 12 may contain reagents 27. For example, in the case of a test for detecting the presence of endotoxin, the well 12 contains three different reagents 27, including a detection reagent that is inactive in the absence of endotoxin activation, an activation reagent (including an enzyme and a fluorescent substrate) for activating the detection reagent, and a control reagent 27 suitable for controlling the functionality of the detection reagent 27. The wells 12 are connected by a channel 13, which is configured to supply a liquid biological sample or another liquid (such as a reference fluid for the control well 12) to the wells 12. The channel 13 is defined by two edges 13a, 13b. Please note that the edges 13a, 13b constitute the interface between the interior of the channel 13 and the face 10a of the plate 9. The edges 13a, 13b extend relative to each other along the direction in which the channel 13 extends. It should be noted that in Figure 3 In FIG. 1 , the edges 13a, 13b cannot be distinguished from the channel 13, which is very thin and is depicted using a line. Figure 8 The channel 13 is present both on the face 10a of the plate 9 and on the edge 11 of the plate 9. In other words, it can be said that the channel 13 extends from the edge 11 to the face 10a, or that the channel 13 extends from the face 10a to the edge 11.

[0056] The board 9 is placed in the receiving space 4 of the base 2. This space (or functional gap) is necessary for the board 9 to be easily inserted into the base 2. More specifically, the board 9 is inserted into the receiving space 4 via the opening 8. The board 9 is arranged so that the edge 11 of the board 9 is arranged to face the first boundary 7 of the base 2. Preferably, the reference Figure 1 , the step b) of placing the plate 9 in the receiving space 4 comprises a step b1), wherein the mouth of the channel 13 of the plate 9 is positioned opposite to the mouth of the duct 5 of the base 2. Thus, each channel 13 opens opposite the mouth of the duct 5, so that each channel 13 forms a fluid passage with the duct 5. Therefore, the plate 9 is considered to be correctly positioned in the receiving space 4 when the opening of the channel 13 is located in the center of the mouth of the duct 5 and thus forms a fluid passage. Therefore, it is preferably ensured that the fluid connection between the channel 13 and the duct 5 is at the edge 11 of the plate 9, which will enable an analytical card 1 that is smaller in volume and easier to package to be obtained. The plate 9 can be automatically inserted into the receiving space 4 using a clamp, which clamps the plate 9 and inserts the plate 9 into the receiving space, and the clamping area of ​​the clamp enters the receiving space 4. As described above, according to a preferred embodiment, the first boundary 7 and the second boundary 230 form a pocket 23 in the base 2. A "pocket" is a receiving space whose border 7, 230 is longer than the opening, more specifically a container open only on one side of the base 2, formed by the film 3 and bounded by the first border 7 and the second border 230. Thus, the arrangement of the plate 9 consists in inserting the plate 9 into the pocket 23. Preferably, the pocket 23 is fitted so that the plate 9 does not flip or change direction if the base 2 equipped with the plate 9 is moved. In other words, the first border 7 and the second border 230 surround the plate 9, except along the opening 8, so that when the plate 9 is arranged in the receiving space 4, almost no receiving space 4 is left empty.

[0057] Next comes a welding step c), which includes welding the two films 3 of the receiving space 4 to the plate 9. Preferably, step c) includes welding the films 3 to the face 10 of the plate 9. Therefore, the two films 3 are welded to the faces 10a, 10b of the plate 9 in the region of the receiving space 4. In other words, the first film 3a is welded to the face 10a, and the second film 3b is welded to the face 10b. The welding can be performed using a heated object that melts the films 3 on the plate 9. The films 3 are thereby welded to the faces 10a, 10b of the plate 9. For example, the heated object may have a face whose size is at least equal to one of the sizes of the faces 10a, 10b. When pressed on the face 10a or 10b, the face of the heated object melts the films 3a, 3b arranged on the face 10a or 10b, and the films 3a, 3b become welded to the face 10a or 10b. After the welding is completed, it can be seen that the plate 9 is fixed relative to the base 2. The plate 9 no longer moves in the receiving space 4.

[0058] Advantageously, the welding of the film 3a to the face 10a and the welding of the film 3b to the face 10b are carried out simultaneously. This allows the method to be improved in many aspects. First, the welding step is carried out faster. Second, the plate 9 arranged in the base 2 is only placed in the welding machine once. If step c) includes two steps of welding the film 3 to the face 10 of the plate 9 (one step for the first film 3a and one step for the second film 3b), once one film 3 has been welded to one face 10, it is necessary to flip the plate 9 and the base 2 so that another film 3 is welded to another face 10. Placing the plate 9 and the base 2 in the welding machine only once makes it possible to speed up the method and limit potential positioning errors. Third, since the plate 9 and the base 2 are positioned in exactly the same way so that the film 3a is welded to the face 10a, and the film 3b is welded to the face 10b (because these two weldings are carried out simultaneously), the welding will be more uniform. In order to weld two films 3 simultaneously on the face 10 of the plate 9, two heating objects need to be applied simultaneously, wherein the first heating object is applied to the first film 3a and the first face 10a, and the second heating object is applied to the second film 3b and the second face 10b. According to one embodiment, the heating objects are made of nickel-chromium alloy and are not heated before being applied to the films 3a, 3b and the faces 10a, 10b. When used, these nickel-chromium alloy heating objects are first pressed on the films 3a, 3b and the faces 10a, 10b, and then current pulses are passed through the nickel-chromium alloy heating objects, which induces the nickel-chromium alloy heating objects to heat up rapidly.

[0059] According to another embodiment, the welding of the film 3a to the surface 10a and the welding of the film 3b to the surface 10b are performed continuously.

[0060] The welding step c) comprises welding the first film 3a to the edge 11 section of the plate 9 by applying a heating block 14 to said first film 3a against said edge 11 on the side of the first face 10a. The welding of the first film 3a to the edge 11 section of the plate is preferably carried out simultaneously with or after the welding of the first film 3a to the face 10 of the plate 9.

[0061] If the film 3 is only welded to the face 10 of the plate 9, the edge 11 of the plate 9 will remain free, that is, no film 3 will be welded to the edge 11. As a result, the biological samples circulating in the channel 13 will be able to mix with each other at the edge 11, and each biological sample can leave its channel 13 and circulate to another channel 13. As mentioned above, if it is desired to obtain effective and usable reaction results, it is not possible to support such mixing of different biological samples before they reach the reaction holes 12. For example, in the case of endotoxin detection tests, biological samples must be analyzed at different concentrations. These fluids must not be mixed before reaching their respective holes 12, otherwise the results of the reaction will not be usable. Therefore, it is necessary to weld the film 3 to a section of the edge 11 in such a way as to strengthen the connection between the plate 9 and the base 2 and isolate the channel 13.

[0062] Preferably, the section of edge 11 to which film 3a is welded is a separation surface 18 on edge 11 between channels 13, such as Figure 8 As shown. The separation surface 18 is Figure 8 18 is a surface shown as a full dashed line in FIG. 1 . Thus, at the edge 11, the channels 13 are separated by a separation surface 18. Therefore, it should be understood here that the separation surface 18 is a section of the edge 11 and is therefore part of the plate 9. The separation surface 18 has an arbitrary shape and is delimited by at least one second side 180. Preferably, the separation surface 18 has a rectangular shape and is therefore delimited by four sides 180a-180d. The separation surface 18 may be on the edge 11 along Figure 8 The direction x shown extends longitudinally from the edges 13a, 13b of the first channel 13 to the edges 13b, 13a of the second channel 13, which is continuous with the first channel 13. Preferably, the separation surface 18 does not contact the channel 13 and is therefore located between a first boundary fixed at a first distance from the edges 13a, 13b of the first channel 13 and a second boundary fixed at a second distance from the edges 13b, 13a of the second channel 13. In other words, there is a virtual margin around the channel 13, which the separation surface 18 does not break. Figure 8 As shown, the side surfaces 180a and 180c do not contact the edges 130b, 131a of the channels 130, 131. It should be reiterated that the separation surface 18 is preferably rectangular, and therefore, more preferably, as Figure 8As shown, the side surfaces 180a and 180c are parallel to the edges 130b and 131a of the channels 130 and 131. More specifically, as shown in the schematic diagram, the side surface 180a is located on the virtual edge of the edge 130b, and the side surface 180c is located on the virtual edge of the edge 131a. Figure 8 In the y-direction shown, the separation surface 18 does not extend beyond the edge 11. Therefore, the separation surface 18 does not extend as far as the face 10a of the plate 9, nor as far as the face 10b of the plate 9. In other words, the side faces 180b and 180d are therefore located between the interface of the edge 11 with the face 10a and the interface of the edge 11 with the face 10b. Preferably, the side faces 180b and 180d are parallel to the interface of the edge 11 with the face 10a and the interface of the edge 11 with the face 10b. More preferably, the side face 180b coincides with the interface of the edge 11 with the face 10a, and the side face 180d coincides with the interface of the edge 11 with the face 10b.

[0063] use Fig. 9 The heating block 14 shown welds the film 3 to the edge 11 section. The heating block 14 comprises a bracket 15 and a heating element 16. The heating element 16 is heated to a heating temperature suitable for at least partially melting the film 3, for example at least 60°C. The bracket 15 and the heating element 16 can be two independent components connected to each other. For example, the bracket 15 and the heating element 16 can be bonded or welded. The heating element 16 is made of a thermally conductive material, typically made of metal. The heating element 16 comprises a first portion 160, which protrudes along the edge 11 at least during the application of the heating block 14 to the edge 11, substantially protruding from one face 10a to another face 10b. In the example shown, the first portion 160 protrudes from the bracket 15, in this way forming a protruding portion from the bracket 15.

[0064] The first part of the heating element 160 has a profile that is complementary to at least a part of the profile of the edge 11 of the plate 9. Thus, the first part of the heating element 160 matches the shape of a section of the edge 11. If the edge 11 has a beveled shape, the first part of the heating element 160 has a shape that is complementary to a part of the bevel so that a section of the edge 11 can abut against the first part of the heating element 160. Thus, when the heating block 14 is applied to the film 3a arranged on the edge 11 of the plate 9, the film 3a melts due to the heat from the heating element 16 and begins to weld to the section of the edge 11 relative to which the first part of the heating element 160 has a complementary profile. Preferably, the profile of the first part of the heating element 160 is complementary to the separation surface 18 of the edge 11. In other words, the heating block 14 comprises a non-heating area 26, the width of which is greater than the width of the channel 13. The non-heating area 26 can be recesses 26 in the first part of the heating element 16. These recesses 26 can, for example, be in the form of grooves. The non-heating zone 26 may also be a portion of the first portion of the heating element 160 that is made of a non-thermally conductive insulating material. When the heating element 16 is applied to the edge 11 of the plate 9, the non-heating zone 26 is positioned at the channel 13 so that no portion of the heating element 16 is directly applied to the channel 13. Thus, the heating element 16 does not weld the film 3a to the channel 13, but welds the film 3a to the separation surface 18, and thus only between the channels 13 at the edge 11. This makes it possible to clearly isolate the channels 13 from each other and prevents the biological sample from circulating between the multiple channels 13 at the edge 11.

[0065] As in the embodiment shown, the heating element can be rigid, in which case the first portion of the heating element 160 permanently protrudes and always has a contour complementary to the contour of the edge 11. The heating element 16 can also be flexible, and thus the protrusion of the first portion of the heating element 160 can be caused by applying the heating element 16 to the edge 11. For example, the first portion of the heating element 160 can comprise a strip or a bundle of metal wires, for example made of nickel-chromium alloy (NiCr). The flexibility of the heating element 16 allows self-adjustment to compensate for any small geometric differences, such as those generated during the method of manufacturing the plate 9 by injection molding.

[0066] According to a preferred embodiment, the welding of the membrane 3 to the edge 11 section of the plate 9 extends at least as far as the inter-duct boundary 7a separating the ducts 5. It should therefore be understood that the welding of the membrane 3 thus extends continuously from the edge 11 section at least to the inter-duct boundary 7a. In other words, step c) comprises welding the membrane 3a to the transition sub-region 20 of the transition region 19 of the membrane 3b, thereby defining a transition space 25. Reference Figure 3Once the plate 9 is fixed in the base 2, a transition space 25 is formed between the edge 11 and the first border 7. The transition space 25 is defined between the portion of the film 3a and the portion of the film 3b, which is the transition zone 19. Thus, the transition zone 19 is the portion of the film 3b that is not welded to the film 3a or the plate 9 and is located between the edge 11 and the first border 7. Figure 3 In the embodiment, the transition region 19 is a striped surface. The transition region 19 comprises a transition sub-region 20 which is located within the extension of the separation surface 18 and extends as far as the inter-duct boundary 7a. Figure 8 Shows Figure 3 Magnified image of . Figure 8 The transition sub-region 20 is shown in FIG. 1 and is a point-shaped surface within the transition region 19, which itself is in FIG. Figure 8 The transition sub-region 20 is not located opposite to the opening of the channel 13 or the pipe 5 in the transition region 19. Instead, the transition sub-region 20 is located between the fluid paths. As a reminder, the fluid paths are formed by the channels 13 and the pipes 5 that are open to each other. Therefore, each fluid path includes the channel 13, the pipe 5, and the transition region 19 portion, so that the biological sample circulating in the pipe 5 can reach the channel 13 via the transition region 19 portion.

[0067] It should be noted that the transition space 25 is part of the receiving space 4 and is located between the edge 11 of the plate 9 and the first border 7 of the base 2. This transition space 25 poses a problem for the analysis of different biological samples. It is not desirable to leave the transition space 25 free, since this would allow the different biological samples circulating from the duct 5 of the base 2 to the channel 13 of the plate 9 to mix. It must be understood that even if the plate 9 is arranged in such a way as to minimize the surface area of ​​the transition zone 19 and therefore the volume of the transition space 25 before the film 3 is welded to the faces 10a, 10b, i.e. before the interface between the edge 11 and the face 10b of the plate 9 is connected to the first border 7, the transition zone 19 and the transition space 25 will still exist. This is because, firstly, the film 3 is flexible and if the film 3 is only welded to the face of the plate 9, the transition zone 19 and the transition space 25, no matter how narrow, will remain. This can also be explained by the fact that when the film 3 is welded to the faces 10a, 10b, there is no guarantee that the plate 9 will not move inside the receiving space 4, no matter how slightly. Therefore, the transition sub-region 20 of the film 3b is preferably welded to the film 3a. Therefore, it should be understood that the film 3a will be welded to the transition sub-region 20, and thus the films 3a and 3b will be welded to each other between the fluid passages to allow the biological sample to circulate between the pipes 5 and the channels 13 of each fluid passage 26, while preventing the biological sample from circulating between the pipes 5 and the channels 13 of different fluid passages 26. Therefore, the films 3a and 3b are welded together up to the inter-channel boundary 7a, so that the isolation of the multiple fluid passages can be enhanced so that the biological sample circulated from the pipe 5 to the channel 13 is not mixed in the transition region 19. In this preferred embodiment, the first portion of the heating element 160 has a contour having a shape complementary to both the edge 11 section of the plate 9 and the portion of the base 2 in the transition sub-region 20.

[0068] More preferably, the welding of the membrane 3 extends beyond the inter-duct boundary 7a. In other words, the welding step c) comprises welding the membrane 3 to the plate 9 from a section of the edge 11 of the plate 9 until beyond the inter-duct boundary 7a separating the ducts 5. This therefore means that the membranes 3a and 3b are welded to each other between the ducts 5 in the area 21 of the membrane 3b schematically depicted using a grid pattern, as shown in FIG. Figure 8 As shown. Extending the weld beyond the inter-duct boundary 7a makes it possible to ensure that the films 3a and 3b are completely welded to each other from the edge 11 section to the inter-duct boundary 7a. In this preferred embodiment, the first portion of the heating element 160 has a profile that has a shape complementary to both the edge 11 portion of the plate 9 and the portion of the base 2 in the transition sub-region 20 and extends beyond the inter-duct boundary 7a. Preferably, the duct 5 is not heated to avoid damaging the fragile valve.

[0069] Advantageously, the steps of welding the film 3a to the face 10a of the plate 9 and welding the film 3 to the edge 11 section of the plate 9 are performed simultaneously using the same heating block. In this case, the heating element 16 of the heating block 14 comprises a second portion having a shape complementary to the shape of the face 10 of the plate 9. Thus, when the heating element 16 of the heating block 14 is applied to the film 3a covering the face 10a of the plate 9, the second portion of the heating element matches the shape of the face 10a of the plate 9 and welds the film 3a to the face 10a, while the first portion of the heating element 160 welds the film 3a to the edge 11 section of the plate 9. Welding the film 3a to the face 10a while welding the film 3a to the edge 11 section has several advantages. Firstly, it speeds up the method because the steps are performed simultaneously, rather than separately. It also reduces the number of rejects because problems with the machine operation and the positioning of the plate 9 and the base 2 in the machine are less likely to occur in one step than in two steps. Furthermore, since no special tools are needed for welding the film 3a to the face 10a and for welding the film 3a to the edge 11 section, fewer tools are needed, only the heating block 14, which reduces costs. Finally, the plate 9 is heated only once, which reduces the risk of deterioration of the thermosensitive reagent 27 already present in the wells 12.

[0070] More preferably, the welding of the first film 3a to the first face 10a, the welding of the second film 3b to the second face 10b and the welding of the first film 3a to the edge 11 section are performed simultaneously. Therefore, it should be understood that the welding of the first film 3a to the first face 10a and the welding of the first film 3a to the edge 11 section are performed using a single heating block 14. Another heating object can be used to weld the second film 3b to the second face 10b. While the heating object is applied to the second film 10b, the heating block 14 is applied to the first film 10a. According to this embodiment, the method is therefore even faster.

[0071] According to another embodiment, in particular when the plate 9 has a small thickness (i.e. less than 0.5 mm), the profile of the edge 11 of the plate 9 is not inclined relative to the faces 10a, 10b and can be perpendicular to the faces 10a, 10b. In this case, the films 3a, 3b are welded to the edge 11 by rivet heads.

[0072] Preferably, the method comprises a step of closing the receiving space 4 so that it is no longer open to the outside via the opening 8. The closing step is achieved by welding the two films 3 together. The welding can be performed, for example, using a laser or by applying a heated object to the films 3. Figure 4(depicting the analytical card 1 obtained at the end of the method) shows the welding line 40 of the receiving space 4 closing the pocket 23. Firstly, closing the receiving space 4 ensures that the plate 9 cannot escape from the receiving space 4 and is therefore kept inside the receiving space 4. This also helps to isolate the plate 9 and prevent it from being damaged. It also protects the plate 9 from external influences, thereby preventing any foreign particles or foreign bodies from entering the holes 12 of the plate 9, which would otherwise impair the effectiveness of the reaction in the holes 12 when a biological sample arrives. Preferably, the welding of the two films 3 closing the receiving space 4 tightly surrounds the plate 9 so that the plate 9 is firmly held and fixed inside the base 2. Therefore, preferably, in Figure 3 In the embodiment, the weld closing the opening 8 extends along the side of the plate 9 and more preferably, the distance separating the weld closing the opening 8 from the plate 9 is less than 5 mm, more preferably less than 2 mm.

[0073] The method of manufacturing the analysis card 1 preferably includes a step of placing the plate 9 under vacuum. The step of placing the plate 9 under vacuum is intended to create a vacuum in the wells 12 and channels 13 of the plate 9. When a vacuum is created in the plate 9, any biological sample introduced into the conduit 5 of the base 2 will be sucked into the channel 13 and then enter the wells 12 of the plate 9. Therefore, a reaction will occur between the biological sample in the well 12 and the reagent 28. In addition, before the step of welding the film 3 to the face 10 of the plate 9, a vacuum step may be implemented. Therefore, when the film 3 is welded to the face 10 of the plate 9, there are no bubbles between the film 3 and the face 10.

[0074] To perform this vacuum step, refer to Figure 4 , the supplied plate 9 advantageously has a vacuum port 24. The vacuum port 24 is connected to the channel 13 of the plate 9. Preferably, the vacuum port 24 leads to at least one face 10a of the plate 9. A vacuum member (e.g. a suction cup) is adhered to the vacuum port and creates a vacuum in the channel 13 and the hole 12 of the plate 9. This operation can last from 15 seconds to 2 minutes, typically 30 seconds. Next, a heating member is applied to the channel 13 connected to the vacuum port 24 to melt the plate 9 at the channel 13 and plug the channel 13. Thus, a vacuum is obtained in the hole 12 and the channel 13 of the plate 9.

[0075] According to an embodiment in which the film 3 is already welded to the surface 10 when the vacuum is created, the vacuum step comprises piercing the film 3 covering the vacuum port 24. In particular, the vacuum port 24 must be accessed to suck out the air and create a vacuum inside the plate 9. Furthermore, the duct 5 is preferably not open to the outside in order for the vacuum to be effective. Thus, for example, the duct 5 may comprise a frangible valve that closes the duct 5 at least during the vacuum step.

[0076] The invention is not limited to the embodiments described and shown in the drawings. Modifications may be made, particularly from the point of view of the nature of the various technical features or the substitution of technical equivalents, without departing from the scope of protection of the invention.

Claims

1. A method for manufacturing a bioanalysis card (1) configured to perform a biological analysis on a biological sample, the method comprising: - Step a): Providing a substrate (2) consisting of: - at least two overlapping films (3), - a receiving space (4) formed between the two films (3), which is delimited by a first boundary (7) and an opening (8), a duct (5) delimited by a contour line (6) which opens into the receiving space (4) via a first boundary (7), - Step b): Placing a plate (9) in the receiving space (4) by inserting it into the opening (8), said plate (9) comprising: - a first face (10a) and a second face (10b) connected by an edge (11) having a profile, - a plurality of holes (12) opening into at least the first face (10a) or the second face (10b), - a plurality of channels (13) which fluidically connect the holes (12), - a welding step c) consisting in welding the two films (3) of the receiving space (4) to the plate (9), The method is characterized in that the welding step c) comprises welding the film (3) to a section of the edge (11) of the plate (9) by applying a heating block (14) to the film (3) against the edge (11) on the side of the first face (10a), the heating block (14) comprising a support (15) and a heating element (16), a first part (160) of the heating element protruding from the support (15) at least during the application of the heating block (14), the first part (160) of the heating element having a profile complementary to at least a section of the profile of the edge (11) of the plate (9), so that during the application of the heating block (14), when the film (3) is pressed by the first part (160) of the heating element, the film (3) takes the shape of the edge (11).

2. A method for manufacturing an analytical card (1) as described in claim 1, wherein the profile of the segment of the edge (11) of the plate (9) is a straight line, which defines a convex angle (θ) between the segment of the edge (11) of the plate (9) and the second surface (10b), and the convex angle is less than or equal to 90°, or the profile of the segment of the edge (11) of the plate (9) is a curve, and the tangent of the curve defines a convex angle (θ) between the segment of the edge (11) of the plate (9) and the second surface (10b), and the convex angle is less than or equal to 90°.

3. A method for manufacturing an analytical card (1) according to any preceding claim, wherein the edge (11) section to which the film (3) is welded is a separation surface (18) on the edge (11) between the channels (13).

4. A method for manufacturing an analytical card (1) as claimed in the preceding claim, wherein the first portion (160) of the heating element of the heating block (14) comprises a non-heating zone (26) whose width is greater than the width of the channel (13), so that when the first portion (160) of the heating element is applied to the film (3) against the edge (11) section, the non-heating zone (26) faces the channel (13) and the frangible valve.

5. A method for manufacturing an analytical card (1) as claimed in any one of the preceding claims, wherein step c) comprises welding the film (3) from a section of the edge (11) of the plate (9) to the inter-channel boundaries (7a) separating the channels (5) by means of a heating block (14).

6. Method for manufacturing an analytical card (1) according to any of the preceding claims, wherein step b) comprises the step of positioning the plate (9) in the receiving space (4) until the mouth of the channel (13) is opposite the mouth of the duct (5).

7. Method for producing an analytical card (1) according to any of the preceding claims, wherein step c) comprises the step of closing the opening (8) of the receiving space (4) by welding the two films (3) to each other.

8. A method for manufacturing an analytical card (1) as claimed in any one of the preceding claims, wherein the plate (9) comprises a vacuum port (24) connected to a channel (13) connected to the hole (12), and wherein the method comprises a vacuum step after the welding step c), the vacuum step comprising piercing a film (3) covering the vacuum port (24) and blocking the channel (13) connecting the vacuum port (24) and the hole (12) after the vacuum is generated.

9. Method for manufacturing an analytical card (1) according to any of the preceding claims, wherein the thickness of the plate (9) is greater than 0.5 mm.

10. Method for producing an analytical card (1) according to any of the preceding claims, wherein welding the two films (3) of the receiving space (4) to the plate (9) comprises welding the first film (3a) to the first face (10a).

11. A method for manufacturing an analytical card (1) as claimed in any of the preceding claims, wherein the heating block comprises a second portion of the heating element, the shape of the second portion being complementary to the shape of the first face (10a) of the plate (9), and the welding of the two films (3) of the receiving space (4) to the plate (9) comprises welding the first film (3a) to the first face (10a), the first film (3a) being pressed against the first face (10a).

12. An analytical card (1) produced by the method according to any one of claims 1 to 11.

13. A heating block (14) configured to implement the method of any one of claims 1 to 11, the heating block (14) comprising a support (15) and a heating element (16), a first portion (160) of the heating element being configured to protrude at least during application of the heating block (14), the first portion (160) of the heating element having a profile complementary to at least a portion of a profile of an edge (11) of a plate (9).

14. The heating block (14) of the preceding claim, wherein the first portion (160) of the heating element comprises a non-heating zone (26) having a width greater than the width of the channel (13).

15. The heating block (14) of the preceding claim, wherein the non-heating area (26) of the first portion (160) of the heating element is a groove (26).

16. The heating block (14) of one of the preceding claims, wherein the first portion (160) of the heating element comprises a flexible nickel-chromium alloy (NiCr) element.

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