Method for manufacturing a biological analysis card involving a heating block

Abstract

The invention relates to a method for manufacturing a biological analysis card (1) comprising: - a step a) of providing a base (2) formed of: - at least two films (3) placed one on top of the other; - a receiving space (4) formed between the two films (3); - ducts (5) leading to the receiving space (4); - a step b) of placing a plate (9) in the receiving space (4), the plate (9) comprising: - a first face (10a) and a second face connected by an edge (11) having a profile; - a plurality of wells (12) opening onto at least the first face (10a) or the second face; - a plurality of channels (13) fluidly connecting the wells (12); - a welding step c) for welding the two films (3) in the receiving space (4) to the plate (9) and a film (3) to segments of the edge (11) of the plate (9) by applying a heating block to the film (3) against the edge (11) on the side of the first face (10a).

Description

[0001] DESCRIPTION

[0002] Title: Manufacturing process for a biological analysis card involving a heating block

[0003] Technical field

[0004] The present invention relates to the field of biological sample analysis, and more specifically relates to a method of manufacturing an analysis card using a heating block, the analysis card obtained by the method and the heating block used.

[0005] Technological background

[0006] In vitro biological analysis of biological samples, such as polymerase chain reaction (PCR) tests, relies on one or more reactions between the biological sample and one or more reagents. A biological sample may include tissues and cells from a human or animal body and their derivatives, organs, blood, its components, or its derivatives. Microfluidic systems and methods are available for performing these biological sample analyses. The reagents are placed in wells of a plate called an array, and the biological sample to be analyzed is brought into contact with the reagents by flooding. The plate is assembled to a flexible base that includes biological sample reservoirs and conduits. The conduits of the base are connected to the channels of the plate to form fluid paths.An analysis card, such as the "FilmArray®" analysis card for running PCR tests, is formed. Assembling this card is a complex procedure that requires adhering at least four layers of film and adhesive layers to the plate before attaching the plate to the base. This involves the use of special machines and a large quantity of consumables (film and adhesive layers), which significantly increases costs and increases the risk of defects due to possible leaks between the layers.

[0007] Furthermore, in the configuration of the "FilmArray®" analysis card, only one biological sample arrives in the plate via a conduit. "FilmArray®" cannot therefore be used for the analysis of several biological solutions, for example the same biological sample at different concentrations, within the same analysis card and therefore for example in the case of endotoxin detection tests. Indeed, during 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 biological sample dilutions do not mix with each other before reaching the reaction wells of the plate so that the reaction results are usable.Finally, if we take the example of endotoxin detection tests, a specificity that must be emphasized is that of the thickness of the plate which is much greater than that of the “FilmArray®”. The detection sensitivity required for endotoxins often requires a volume of reagents greater than that necessary for PCR tests, which has an impact on the thickness of the reaction wells and therefore on the thickness of the analysis card which must therefore be greater (the intensity of the detected fluorescence signal being directly proportional to the thickness of fluorescent liquid present in the reaction wells).This particularity involves problems of sealing the plate in the base since sealing the faces of the plate between films of the base as is the case for “FilmArray®” does not ensure sealing at the edges of the plate, and therefore presents a risk of mixing biological samples, or other liquids.

[0008] There is currently no manufacturing process for obtaining an analysis card with a thick plate between two base films, for example in the context of endotoxin detection tests, which is quick, easy to produce and inexpensive.

[0009] Presentation of the invention

[0010] The invention therefore aims to manufacture an analysis card provided with a plate having a determined thickness capable of ensuring the separation of the fluidic pathways extending in the analysis card, for example making it possible to analyze several dilutions of the same biological sample or different biological samples or different biological samples with several dilutions, in a more controlled manner, without risk of mixing, more quickly, more easily and less expensively.

[0011] According to a first aspect, there is provided a method of manufacturing a biological analysis card configured for the biological analysis of a biological sample, said method comprising:

[0012] - a step a) of providing a formed base:

[0013] - at least two superimposed films,

[0014] - a reception space formed between the two films delimited by a first border and an opening,

[0015] - conduits delimited by contours and opening onto the reception area through the first border,

[0016] - a step b) of placing a plate in the reception space by inserting the plate through the opening, the plate comprising:

[0017] - a first face and a second face connected by an edge presenting a profile,

[0018] - a plurality of wells opening onto at least the first face or the second face,

[0019] - a plurality of channels fluidly connecting wells, - a welding step c) comprising welding the two films of the receiving space to the plate, the method being characterized in that the welding step c) comprises welding a film to segments of the edge of the plate by applying a heating block to said film against said edge on the side of the first face, said heating block comprising a support and a heating element, a first part of the heating element projecting from the support at least during the application of the heating block, said first part of the heating element then having a profile complementary to at least one segment of the profile of the edge of the plate, so that during the application of the heating block, the film takes the shape of the edge when said film is pressed by the first part of the heating element.

[0020] According to advantageous and non-limiting characteristics, taken alone or in any combination:

[0021] - the profile of a segment of the edge of the plate is a straight line which defines a salient angle between said segment of the edge of the plate and the second face which is less than or equal to 90° or a curve of which a tangent defines a salient angle between said segment of the edge of the plate and the second face which is less than or equal to 90°;

[0022] - the edge segments to which the film is welded are separation surfaces located on the edge between the channels;

[0023] - the first part of the heating element of the heating block comprises non-heating zones whose width is greater than the width of the channels, so that, when applying the first part of the heating element to the film against the edge segments, the non-heating zones face the channels and fragile valves;

[0024] - each conduit is equipped with at least one fragile valve positioned at one end of the conduit

[0025] - step c) comprises the welding by the heating block of a film from the segments of the edge of the plate to inter-duct borders separating the ducts;

[0026] - step b) comprises a step of positioning the plate in the receiving space until the mouths of the channels are opposite the mouths of the conduits;

[0027] - step c) comprises a step of closing the opening of the reception space by welding the two films to each other;

[0028] - the plate comprises a vacuum orifice connected to channels, said channels being connected to wells and in which the method comprises a vacuum step following the welding step c), the vacuum step comprising piercing the film covering the vacuum orifice and plugging the channels connecting the vacuum orifice to the wells once the vacuum has been created;

[0029] - the thickness of the plate is greater than 0.5 mm; - the welding of the two films of the reception space to the plate comprises the welding of a first film to the first face;

[0030] - the heating block comprises a second part of the heating element whose shape is complementary to that of the first face of the plate, the welding of the two films of the reception space to the plate comprises the welding of a first film to the first face, the first film being pressed onto the first face.

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

[0032] According to a third aspect, a heating block is provided configured to implement the method presented above, said heating block comprising a support and a heating element, a first part of the heating element being configured to protrude at least during the application of the heating block, said first part of the heating element then having a profile complementary to at least part of the profile of the edge of the plate.

[0033] The heating block is advantageously complemented by the following different characteristics taken alone or according to their different possible combinations:

[0034] - the first part of the heating element comprises non-heating zones whose width is greater than the width of the channels;

[0035] - the non-heating areas of the first part of the heating element are recesses;

[0036] - the first part of the heating element includes flexible nichrome (NiCr) elements.

[0037] Detailed description

[0038] Other characteristics, aims and advantages of the invention will emerge from the following description, which is purely illustrative and non-limiting, and which must be read in conjunction with the appended drawings in which: Figure 1 is a diagram showing steps of the method of manufacturing the analysis card according to a possible embodiment of the invention; Figure 2 schematically illustrates a basic example of an analysis card according to a possible embodiment of the invention; Figure 3 shows an example of an analysis card during manufacture according to a possible embodiment of the invention; Figure 4 schematically illustrates an example of an analysis card according to a possible embodiment of the invention; Figure 5 represents a perspective view of a part of an analysis card according to a possible embodiment of the invention;Figure 6 schematically shows the heating block pressing the first film against the edge of an analysis card plate having a profile forming a straight line according to a possible embodiment of the invention; Figure 7 schematically shows the heating block pressing the first film against the edge of an analysis card plate having a curved profile according to a possible embodiment of the invention; Figure 8 represents an enlarged view of the analysis card at an edge of the plate; Figure 9 illustrates a heating block and a plate.;

[0039] The present invention relates to a method for manufacturing an analysis card 1 illustrated in Figure 4. Figure 1 details the steps of the method which firstly comprises a step a) of providing a base 2. With reference to Figure 2, the base 2 is formed from two superimposed thermoplastic films 3. For example, the constituent material of the films 3 may be a polypropylene-based complex or polyethylene. The films 3 are thin, soft and flexible surfaces. By "thin", it is meant that the thickness of the films 3 is less than 200 μm. Preferably, the films 3 have a rectangular shape. More preferably, the two films 3a, 3b have the same shape when superimposed. Thus, it is not possible to distinguish the two films 3a, 3b in Figure 2 since they are one on top of the other. The film 3b is referenced in Figure 5.The two films 3 are thermoplastic, which means that by heating them up to a certain temperature threshold, the films 3 will soften, which will allow them to be subjected to a mechanical deformation which is fixed when the films 3 cool. The temperature threshold from which the films 3 soften and can be deformed depends on the material constituting the films 3. The material constituting the films 3 is preferably a polymer material such as polypropylene or polyethylene or a complex film based on polypropylene or polyethylene. Typically, the temperature threshold necessary to soften the films 3 is greater than 60°C.

[0040] A receiving space 4 is formed between the two films 3. More precisely, the receiving space 4 is comprised between two free film portions 3. By "free" is meant 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 edge 7 and an opening 8. The first edge 7 is an area in which the two films 3 are welded together. In the example illustrated in Figure 2, the first edge 7 forms a straight line parallel to the width of the films 3 and, along this straight line, the two films 3 are welded to each other. This welding can for example be implemented using a laser which melts the films 3 along a precise trajectory so that, along this trajectory, the two films 3 are welded to each other. Other borders where the two films 3 are welded together can delimit the reception space 4.The opening 8 corresponds to an area in which the films 3 are not welded together, and are left free up to their 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 intended to accommodate, typically rectangular.

[0041] The opening 8 may comprise one or more of the three different edge portions of the films 3 completing the first border 7, and located on the right in Figure 2. In this case, the opening 8 therefore comprises an edge portion of the films 3 parallel to the first border 7 and two edge portions of the films 3 perpendicular to the first border 7. Each of these edge portions of the films 3 is free and is not welded to an edge portion of another film 3. According to a preferred embodiment illustrated in Figure 2, the first border 7 and a second border 230 of the base 2 form a pocket 23 such that the opening 8 is located on only one side of a film 3.

[0042] The base 2 further comprises conduits 5 formed between the two films 3 and delimited by contours 6. A conduit 5 can be likened to a tunnel formed between the two films 3 which allows the circulation of fluid such as a liquid biological sample. The contours 6 of the conduits 5 form curves along which the two films 3 are welded to each other, in the same way as the first border 7. Each conduit 5 is delimited by two contours 6. The contours 6 of different conduits 5 are connected by inter-conduit borders 7a, in which the two films 3 are welded to each other. The inter-duct borders 7a are part of the first border 7. The base 2 may also comprise various elements such as reservoirs formed between the films of the base 2 and connected to the conduits 5. The conduits 5 may also each comprise one or more fragile valves blocking said conduits 5.For example, these fragile valves can be defined by an area where the films 3 are weakly welded (a few millimeters or a few centimeters), and are configured to break depending on a pressure that would be applied to said fragile valves. A fragile valve is preferably located close to the receiving space 4, that is to say at the end of a conduit 5 which opens onto the receiving space 4. For example, the reservoirs, which can have the shape of a shell or "blister" in English, can contain the biological sample. The reservoirs can be pressed until the pressure applied to the fragile valves present in the conduits 5 connected to the reservoirs reaches a certain pressure threshold necessary for the rupture of the fragile valves. Once the fragile valves are ruptured, the biological samples contained in the reservoirs flow into the conduits 5 to the receiving space 4.

[0043] The manufacturing method then comprises a step b) of arranging a plate 9 in the base 2 so as to result in a base 2 equipped with a plate 9 as illustrated in Figure 3. A plate 9 is generally defined as a flat surface element 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 less than the widths and lengths of the faces. Preferably, the plate 9 has a thickness greater than or equal to 0.5 mm. In the example illustrated, the plate 9 has a rectangular surface, but other shapes could be chosen.

[0044] Preferably, the plate 9 is formed from a thermoplastic material, at least at its surfaces. For example, the plate 9 can be made from materials such as PP, PE, PMMA, PC, PS, POM, ABS, COP. As shown schematically in Figure 5, the plate 9 is composed of two faces 10a, 10b opposite each other and separated by a small thickness, i.e. the thickness is at least 10 times less than the widths and lengths of the faces 10a, 10b. The largest dimensions and the smallest dimensions of the faces 10a, 10b of the plate 9, in this case the lengths and widths of the faces 10a, 10b, are preferably greater than 2 cm and, also preferably, less than 10 cm, and, 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 sides, including an edge 11.In other words, the edge 11 corresponds to another face of the plate 9, different from the faces 10a and 10b, and whose width is not very significant, i.e. at least 10 times less than the widths and lengths of the faces 10a, 10b. The length of the edge 11 is therefore equal to a length or a width of the faces of the faces 10a, 10b of the plate 9. Preferably, the profile of the edge 11 is inclined relative to the faces 10a, 10b and is therefore not perpendicular to the faces 10a, 10b. Preferably, with reference to Figure 6, the profile of the edge 11 is a straight line which defines a salient angle 0 between the edge 11 of the plate 9 and the second face 10b which is less than 90°. In other words, the smaller of the two angles defined by the profile of the edge 11 between the edge 11 and the second face 10b, i.e. the one which is less than 180°, is less than 90°. In other words, the edge 11 preferably forms a bevel.According to another embodiment illustrated in Figure 7, the profile of the edge 11 is a curve of which a tangent defines a salient angle 0 between the edge 11 of the plate 9 and the second face 10b which is less than 90°. A tangent is for example illustrated in dotted lines in Figure 7. In other words, the smaller of the two angles defined by the profile of the edge 11 between the edge 11 and the second face 10b, i.e. the one which is less than 180°, is less than 90°. According to this embodiment, the profile of the edge 11 is preferably a convex curve.

[0045] The plate 9 comprises a plurality of wells 12 opening at least onto one face 10a, 10b of the plate 9. The wells 12 can also pass through the plate 9 from one face 10a to another face 10b. The wells 12 can contain reagents 27. For example, in the case of tests for detecting the presence of endotoxins, the wells 12 contain three different reagents 27 comprising a detection agent in a non-active state in the absence of activation free of endotoxins, an agent for activating the detection agent comprising an enzyme and a fluorogenic substrate, and a control reagent 27 adapted to control the functionality of the detection reagent 27. The wells 12 are connected by channels 13 configured to supply the wells 12 with a liquid biological sample or another liquid such as a reference fluid used for example for control wells 12. The channels 13 are delimited by two edges 13a, 13b.It is understood that the edges 13a, 13b constitute the interface between the interior of a channel 13 and the face 10a of the plate 9. The edges 13a, 13b extend opposite each other in the direction in which a channel 13 extends. It is specified that, in Figure 3, the edges 13a, 13b are not distinguished from the channels 13, the channels 13 being very thin and being represented with lines. The edges 13a, 13b are referenced in Figure 8. The channels 13 are 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 stated that the channels 13 extend from the edge 11 to the face 10a or that the channels 13 extend from the face 10a to the edge 11.

[0046] The plate 9 is placed in the receiving space 4 of the base 2. This space (or functional clearance) is necessary for the easy insertion of the plate 9 into the base 2. More precisely, the plate 9 is inserted into the receiving space 4 via the opening 8. The plate 9 is arranged such that the edge 11 of the plate 9 is arranged opposite the first edge 7 of the base 2. Preferably, with reference to Figure 1, step b) of placing the plate 9 in the receiving space 4 comprises a step b1) in which the mouths of the channels 13 of the plate 9 are positioned opposite the mouths of the conduits 5 of the base 2. Thus, each channel 13 opens opposite the mouth of a conduit 5 so that each channel 13 forms a fluid path with a conduit 5.Consequently, it is considered that the plate 9 is correctly positioned in the receiving space 4 when the mouths of the channels 13 are centered on the mouths of the conduits 5 and the fluidic paths are thus formed. The fluidic connection between a channel 13 and a conduit 5 is therefore preferably ensured at the edge 11 of the plate 9, which will make it possible to obtain a less bulky analysis card 1 that is easier to package. The plate 9 can be inserted automatically into the receiving space 4 using a clamp that grips the plate 9 and inserts the plate 9 into the receiving space, the gripping zones of the clamp engaging in the receiving space 4. As explained previously, according to a preferred embodiment, the first edge 7 and the second edge 230 form a pocket 23 in the base 2.By pocket is meant a receiving space whose edges 7, 230 are longer than the opening, and more precisely a container open on only one side of the base 2 and formed by the films 3 and delimited by the first edge 7 and the second edge 230. The arrangement of the plate 9 therefore comprises the insertion of the plate 9 into the pocket 23. Preferably, the pocket 23 is adjusted in such a way that the plate 9 cannot turn over or change orientation if the base 2 equipped with the plate 9 were to be set in motion. In other words, the first edge 7 and the second edge 230 frame the plate 9, except along the opening 8, so that, when the plate 9 is arranged in the receiving space 4, little empty receiving space 4 remains.

[0047] Next comes a welding step c) comprising the welding of the two films 3 of the receiving space 4 to the plate 9. Preferably, step c) comprises the welding of the films 3 on the faces 10 of the plate 9. The two films 3 are therefore welded at the receiving space 4 to the faces 10a, 10b of the plate 9. In other words, a first film 3a is welded to the face 10a and a second film 3b is welded to the face 10b. This welding can be carried out by heating objects which melt the films 3 on the plate 9. The films 3 are thus welded to the faces 10a, 10b of the plate 9. For example, a heating object can have a face with dimensions at least equal to those of one of the faces 10a, 10b. When pressed against a face 10a or 10b, the face of the heating object melts the film 3a, 3b arranged on the face 10a or 10b and the film 3a, 3b welds to the face 10a or 10b. At the end of this welding, it is understood that the plate 9 is fixed relative to the base 2.Plate 9 can no longer move within reception space 4.

[0048] 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 makes it possible to improve the process in various aspects. Firstly, the welding step is implemented more quickly. Secondly, the plate 9 arranged in the base 2 is positioned only once in the welding machine. If step c) comprises two steps of welding the films 3 to the faces 10 of the plate 9 (one for the first film 3a and one for the second film 3b), the plate 9 and the base 2 must be turned over once a film 3 has been welded to one face 10 to weld the other film 3 to the other face 10. Positioning the plate 9 and the base 2 only once in the welding machine makes it possible to speed up the process and limit potential positioning errors.Third, the welding will be more homogeneous since the plate 9 and the base 2 are positioned in exactly the same way for the welding of the film 3a to the face 10a and the welding of the film 3b to the face 10b (since these two welds are implemented at the same time). To implement the welding of the two films 3 on the faces 10 of the plate 9 simultaneously, two heating objects are applied simultaneously, a first heating object being applied to the first film 3a and the first face 10a and a second heating object being applied to the second film 3b and the second face 10b. According to a certain embodiment, the heating objects are made of nichrome and are not heated before their application to the films 3a, 3b and the faces 10a, 10b.In use, these nichrome heating objects are first pressed the films 3a, 3b and the faces 10a, 10b then a current pulse is sent to the nichrome heating objects which induces rapid heating of the nichrome heating objects.

[0049] According to another embodiment, the welding of the film 3a to the face 10a and the welding of the film 3b to the face 10b are carried out successively.

[0050] Welding step c) comprises welding the first film 3a to edge segments 11 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. Welding the first film 3a to edge segments 11 of the plate

[0051] 9 is preferably implemented simultaneously or after welding the first film 3a on the faces

[0052] 10 of the plate 9. If the films 3 are welded only to the faces 10 of the plate 9, the edge 11 of the plate 9 would remain free, i.e. no film 3 would be welded to the edge 11. Consequently, the biological samples which would circulate in the channels 13 could mix with each other at the edge 11, each biological sample being able to leave its channel 13 and circulate to another channel 13. As explained previously, this mixing of different biological samples prior to their arrival in the reaction wells 12 is unthinkable if valid and usable reaction results are to be obtained. For example, in the case of endotoxin detection tests, a biological sample must be analyzed according to different concentrations. These fluids must not mix before reaching their respective wells 12, otherwise the reaction results cannot be used.Therefore, it is necessary to weld a film 3 to segments of the edge 11 so that the plate 9 is better fixed to the base 2 and the channels 13 are insulated.

[0053] Preferably, the edge segments 11 to which the film 3a is welded are separation surfaces 18 located on the edge 11 between the channels 13 as illustrated in Figure 8. A separation surface 18 is illustrated in Figure 8 by a surface filled with dashes. At the edge 11, the channels 13 are therefore separated by the separation surfaces 18. It will therefore be understood here that the separation surfaces 18 are edge segments 11 and therefore parts of the plate 9. The separation surfaces 18 have any shape and are delimited by at least one second side 180. Preferably, the separation surfaces 18 have a rectangular shape and are therefore delimited by four sides 180a-180d. A separation surface 18 may extend lengthwise, in the direction x indicated in Figure 8, on the edge 11, from an edge 13a, 13b of a first channel 13 to an edge 13b, 13a of a second channel 13, the second channel 13 being consecutive to the first channel 13.Preferably, the separating surfaces 18 are not in contact with a channel 13 and are thus located between a first limit set at a first distance from an edge 13a, 13b of a first channel 13 and a second limit set at a second distance from an edge 13b, 13a of a second channel 13. In other words, there are virtual margins around the channels 13 that the separating surfaces 18 do not cross. As illustrated in Figure 8, the sides 18a and 18c are not in contact with the edges 130b, 131a of the channels 130, 131. It is repeated that, preferably, the separating surfaces 18 are rectangular and thus, still preferably, as illustrated in Figure 8, the sides 180a and 180c are parallel to the edges 130b, 131a of the channels 130, 131. More precisely, on the diagram, side 180a is located on a virtual margin of edge 130b and side 18c is located on a virtual margin of edge 131a.Now regarding the direction y illustrated in Figure 8, the separation surfaces 18 do not extend beyond the edge 11. The separation surfaces 18 therefore do not extend onto the face 10a of the plate 9, nor onto the face 10b of the plate 9. In other words, the sides 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 sides 180b and 180d are parallel to the interface of the edge 11 with the face 10a and to the interface of the edge 11 with the face 10b. More preferably, the side 180b is merged with the interface of the edge 11 with the face 10a and the side 180d is merged with the interface of the edge 11 with the face 10b.

[0054] The welding of a film 3 to the edge segments 11 is implemented by means of a heating block 14 illustrated in Figure 9. The heating block 14 comprises a support 15 and a heating element 16. The heating element 16 is brought to a heating temperature suitable for at least partially melting a film 3, for example at least 60°C. The support 15 and the heating element 16 may be two independent parts connected to each other. For example, the support 15 and the heating element 16 may be glued or welded. The heating element 16 is made of a material that conducts heat, and is typically metallic. The heating element 16 comprises a first part 160 which projects along the edge 11, substantially from one face 10a to the other face 10b, at least during the application of the heating block 14 to the edge 11. In the example illustrated, the first part 160 projects from the support 15 so as to form a rim from the support 15.

[0055] The first part of the heating element 160 has a profile complementary to at least part of the profile of the edge 11 of the plate 9. Consequently, the first part of the heating element 160 follows the shape of segments of the edge 11. If the edge 11 has a bevel shape, the first part of the heating element 160 then has a shape complementary to parts of this bevel so that segments of the edge 11 can fit 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 of the heating element 16, and welds to the edge segments 11 of 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 surfaces 18 of the edge 11.In other words, the heating block 14 comprises non-heating zones 26 whose width is greater than the width of the channels 13. The non-heating zones 26 may be recesses 26 in the first part of the heating element 16. These recesses 26 may, for example, take the form of grooves. The non-heating zones 26 may also be parts made of an insulating material, which does not conduct heat, in the first part of the heating element 160. When the heating element 16 is applied to the edge 11 of the plate 9, the non-heating zones 26 are positioned at the channels 13 in such a way that no part of the heating element 16 is applied directly to the channels 13. Consequently, the heating element 16 does not weld the film 3a to the channels 13 but welds the film 3a to the separation surfaces 18 and therefore only between the channels 13 of the edge 11.This makes it possible to precisely isolate the channels 13 from each other and to avoid the circulation of biological sample between the different channels 13 at the edge 11. As in the illustrated embodiment, the heating element can be rigid, in which case the first part of the heating element 160 protrudes permanently and always has a profile complementary to the profile of the edge 11. It is also possible for the heating element 16 to be flexible, and therefore for the protrusion of the first part of the heating element 160 to be a consequence of the application of said heating element 16 against the edge 11. For example, the first part of the heating element 160 can comprise strips or a bundle of metal wires, for example made of nichrome (NiCr). The flexibility of the heating element 16 allows self-adjustment compensating for possible small geometric differences, for example generated during the manufacturing process by injection of the plate 9.

[0056] According to a preferred embodiment, the welding of a film 3 to segments of the edge 11 of the plate 9 extends at least to the inter-duct edges 7a separating the ducts 5. It is therefore understood that the welding of the film 3 thus extends continuously from the edge segments 11 at least to the inter-duct edges 7a. In other words, step c) comprises welding the film 3a to transitional sub-zones 20 of a transitional zone 19 of the film 3b delimiting a transitional space 25. With reference to Figure 3, once the plate 9 is fixed in the base 2, a transitional space 25 is formed between the edge 11 and the first edge 7. The transitional space 25 is defined between a portion of film 3a and a portion of film 3b, the portion of film 3b being the transitional zone 19. The transitional zone 19 is therefore a portion of film 3b not welded to the film 3a or to the plate 9, between the edge 11 and the first edge 7. In Figure 3, the transitional zone 19 is the scratched surface.The transitional zone 19 comprises transitional sub-zones 20 located in the extension of the separation surfaces 18 and which extend to the inter-duct borders 7a. Figure 8 shows an enlarged view of Figure 3. A transitional sub-zone 20 is illustrated in Figure 8 and is the dotted surface in the transitional zone 19 which is itself striped in Figure 8. The transitional sub-zones 20 are not located opposite mouths of channels 13 or conduits 5 in the transitional zone 19. On the contrary, the transitional sub-zones 20 are located between the fluid paths. It is recalled that a channel 13 forms with a conduit 5, when each opens opposite the other, a fluid path. Each fluid path therefore comprises a channel 13, a conduit 5 and a portion of transitional zone 19 so that the biological sample which circulates in the conduit 5 can reach the channel 13 via this portion of transitional zone 19.

[0057] It is understood that the transitional space 25 is a part of the reception space 4, which is located between the edge 11 of the plate 9 and the first edge 7 of the base 2. This transitional space 25 constitutes a problem for the analysis of different biological samples. It is not desired to leave the transitional space 25 free since it allows the mixing of different biological samples circulating from the conduits 5 of the base 2 to the channels 13 of the plate 9. It must be understood that, even if the plate 9 is arranged in such a way as to reduce as much as possible the surface area of ​​the transitional zone 19 and therefore the volume of the transitional space 25 before welding the films 3 to the faces 10a, 10b, i.e. attaching the interface between the edge 11 and the face 10b of the plate 9 to the first edge 7, the transitional zones 19 and the transitional space 25 will still exist.This is explained firstly by the fact that the films 3 are flexible and that the transitional zone 19 and the transitional space 25, however narrow they may be, remain if the films 3 are welded to only the faces of the plate 9. This is also explained by the fact that, when welding the films 3 to the faces 10a, 10b, it cannot be guaranteed that the plate 9 will not move, even very slightly, within the receiving space 4. Consequently, it is preferable to weld the transitional sub-zones 20 of the film 3b to the film 3a. It is therefore understood that the film 3a will be welded to the transitional sub-zones 20, and therefore that the films 3a and 3b will be welded to each other between the fluid channels, to allow the circulation of biological sample between the conduit 5 and the channel 13 of each fluid channel 26 while preventing the circulation of biological sample between the conduits 5 and the channels 13 of different fluid channels 26.Thus, welding the films 3a and 3b to each other up to the inter-duct edges 7a makes it possible to improve the insulation of the different fluidic paths so that the biological samples which circulate from the ducts 5 to the channels 13 do not mix at the level of the transitional zone 19. In this preferred embodiment, the first part of the heating element 160 has a profile whose shape is complementary to both the edge segments 11 of the plate 9 and parts of the base 2 at the level of the transitional sub-zones 20.

[0058] More preferably, the welds of the films 3 extend beyond the inter-duct edges 7a. In other words, the welding step c) comprises welding a film 3 to the plate 9 from the edge segments 11 of the plate 9 to beyond the inter-duct edges 7a separating the ducts 5. This therefore means that the films 3a and 3b are welded to each other between the ducts 5 as illustrated in Figure 8 at the area 21 of the film 3b shown schematically having a grid pattern. Extending the welding to beyond the inter-duct edges 7a ensures that the films 3a and 3b are perfectly welded to each other from the edge segments 11 to the inter-duct edges 7a.In this preferred embodiment, the first part of the heating element 160 has a profile whose shape is complementary to both edge parts 11 of the plate 9 and parts of the base 2 at the level of the transitional sub-zones 20 up to beyond the inter-duct borders 7a. Preferably, the ducts 5 are not heated so as not to break the fragile valves.

[0059] Advantageously, the step of welding the film 3a to the face 10a of the plate 9 and the step of welding a film 3 to edge segments 11 of the plate 9 are implemented simultaneously by means of the same heating block. In this case, the heating element 16 of the heating block 14 comprises a second part whose shape is complementary to that of a face 10 of the plate 9. Consequently, when the heating element 16 of the heating block 14 is applied to the film 3a which covers the face 10a of the plate 9, the second part 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 part of the heating element 160 welds the film 3a to edge segments 11 of the plate 9. Welding the film 3a to the face 10a simultaneously with welding the film 3a to edge segments 11 has several advantages. First of all, it speeds up the process, the steps being implemented at the same time and not separately.Furthermore, this reduces the number of rejects since problems with the operation of the machine 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. In addition, fewer tools are required since there is no need for a tool dedicated to welding the film 3a to the face 10a and a tool dedicated to welding the film 3a to edge segments 11, there is only the heating block 14, which reduces costs. Finally, the plate 9 is thus heated only once, which reduces the risks of degradation of reagents 27 which would already be present in the wells 12 and which would be sensitive to heat.

[0060] 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 edge segments 11 are carried out simultaneously. Consequently, it is understood that the welding of the first film 3a to the first face 10a and the welding of the first film 3a to edge segments 11 are implemented by means of a single heating block 14. The welding of the second film 3b to the second face 10b can be implemented by means of another heating object. The heating block 14 is applied to the first film 10a simultaneously with the application of a heating object to the second film 10b. According to this embodiment, the method is thus even faster.

[0061] According to another embodiment, in particular in the case where the plate 9 has a low 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 may be perpendicular to the faces 10a, 10b. In this case, the film(s) 3a, 3b are welded at the edge 11 by riveting.

[0062] Preferably, the method comprises a step of closing the receiving space 4 such that the receiving space 4 is no longer open to the outside via the opening 8. The closing step is implemented by welding the two films 3 together. The welding can be carried out for example with a laser or by applying a heating object to the films 3. Figure 4, which illustrates an analysis map 1 obtained at the end of the present method, shows the weld line 40 closing the receiving space 4 of the pocket 230. Closing the receiving space 4 firstly ensures that the plate 9 does not leave the receiving space 4 and therefore remains inside the receiving space 4. Also, this makes it possible to isolate the plate 9 and prevent it from being damaged.Furthermore, this protects the plate 9 from the outside, which prevents the introduction of any foreign particles or foreign objects into the wells 12 of the plate 9 that would threaten the validity of the reactions in the wells 12 upon arrival of biological samples. Preferably, the weld of the two films 3 that closes the receiving space 4 surrounds the plate 9 closely so that the plate 9 is well held and fixed within the base 2. Thus, preferably, in Figure 3, the weld that closes the opening 8 runs along the sides of the plate 9 and, even more preferably, the distance between the weld that closes the opening 8 and the plate 9 is less than 5 mm, and even more preferably less than 2 mm.

[0063] The method for manufacturing the analysis card 1 preferably comprises a step of placing the plate 9 under vacuum. The step of placing the plate 9 under vacuum aims to create a vacuum in the wells 12 and the channels 13 of the plate 9. The vacuum being created in the plate 9, any biological sample which will be introduced into the conduits 5 of the base 2 will be sucked into the channels 13 then into the wells 12 of the plate 9. The reactions between the biological sample and the reagents 28 of the wells 12 will thus take place. Furthermore, a vacuum step can be implemented prior to the step of welding the films 3 to the faces 10 of the plate 9. Thus, when the films 3 are welded to the faces 10 of the plate 9, there are no air bubbles between the films 3 and the faces 10.

[0064] To carry out this vacuuming step, with reference to Figure 4, the plate 9 provided advantageously has a vacuum orifice 24. The vacuum orifice 24 is connected to the channels 13 of the plate 9. Preferably, the vacuum orifice 24 opens onto at least one face 10a of the plate 9. A vacuum member, such as a suction cup, adheres to the vacuum orifice and creates the vacuum in the channels 13 and the wells 12 of the plate 9. The operation can last between 15 seconds and 2 minutes, generally 30 seconds. Then, a heating member is applied to the channels 13 connected to the vacuum orifice 24 to melt the plate 9 at the channels 13 and plug the channels 13. The vacuum is consequently captured in the wells 12 and the channels 13 of the plate 9.

[0065] According to the embodiment in which the films 3 are already welded to the faces 10 during vacuuming, the vacuuming step comprises piercing the film 3 covering the vacuuming orifice 24. Indeed, it is essential to access the vacuuming orifice 24 to suck in the air and create the vacuum within the plate 9. In addition, it is preferable that the conduits 5 are not open to the outside for the vacuuming to be effective. Thus, for example, the conduits 5 may comprise fragile valves which close the conduits 5 at least during the vacuuming step.

[0066] The invention is not limited to the embodiment described and shown in the attached figures. Modifications remain possible, in particular from the point of view of the constitution of the various technical characteristics or by substitution of technical equivalents, without departing from the scope of protection of the invention.

Claims

DEMANDS 1. A method for manufacturing a biological analysis card (1) configured for the biological analysis of a biological sample, said method comprising: - a step a) of providing a base (2) formed: - of at least two superimposed films (3), - a reception area (4) formed between the two films (3) delimited by a first border (7) and an opening (8), - of conduits (5) delimited by contours (6) and opening onto the reception area (4) through the first border (7), - a step b) of installing a plate (9) in the reception area (4) by inserting the plate (9) through the opening (8), the plate (9) comprising: - a first face (10a) and a second face (10b) connected by an edge (11) presenting a profile, - a plurality of shafts (12) opening onto at least the first face (10a) or the second face (10b), - a plurality of channels (13) fluidly connecting wells (12), - a welding step (c) comprising welding the two films (3) of the receiving space (4) to the plate (9), the process being characterized in that the welding step (c) comprises welding a film (3) to segments of the edge (11) of the plate (9) by applying a heating block (14) to said film (3) against said edge (11) on the side of the first face (10a), said heating block (14) comprising a support (15) and a heating element (16), a first part of the heating element (160) protruding from the support (15) at least during the application of the heating block (14), said first part of the heating element (160) then having a profile complementary to at least one segment of the profile of the edge (11) of the plate (9), so that during the application of the heating block (14), the film (3) conforms to the shape of the edge (11) when said film (3) is pressed by the first part of the heating element (160).

2. Method of manufacturing an analysis card (1) according to claim 1, wherein the profile of a segment of the edge (11) of the plate (9) is a straight line which defines a salient angle (0) between said segment of the edge (11) of the plate (9) and the second face (10b) which is less than or equal to 90° or a curve whose tangent defines a salient angle (0) between said segment of the edge (11) of the plate (9) and the second face (10b) which is less than or equal to 90°.

3. Method of manufacturing an analysis card (1) according to any one of the preceding claims, wherein the edge segments (11) to which the film (3) is welded are separation surfaces (18) located on the edge (11) between the channels (13).

4. Method of manufacturing an analysis card (1) according to the preceding claim, wherein the first part of the heating element (160) of the heating block (14) comprises non-heating areas (26) whose width is greater than the width of the channels (13), such that, when the first part of the heating element (160) is applied to the film (3) against the edge segments (11), the non-heating areas (26) face the channels (13) and fragile valves.

5. Method of manufacturing an analysis card (1) according to any one of the preceding claims, wherein step c) comprises welding by the heating block (14) a film (3) from the edge segments (11) of the plate (9) to inter-duct borders (7a) separating the ducts (5).

6. Method of manufacturing an analysis card (1) according to any one of the preceding claims, wherein step b) includes a step of positioning the plate (9) in the receiving space (4) until the mouths of the channels (13) are facing the mouths of the conduits (5).

7. Method of manufacturing an analysis card (1) according to any one of the preceding claims, wherein step c) includes a 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 analysis card (1) according to any one of the preceding claims, wherein the plate (9) comprises a vacuum orifice (24) connected to channels (13), said channels (13) being connected to wells (12) and wherein the method comprises a vacuum step following step c) of welding, the vacuum step comprising piercing the film (3) covering the vacuum orifice (24) and plugging the channels (13) connecting the vacuum orifice (24) to the wells (12) once the vacuum has been achieved.

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

10. Method of manufacturing an analysis card (1) according to any one of the preceding claims, wherein the welding of the two films (3) of the host space (4) to the plate (9) comprises the welding of a first film (3a) to the first face (10a).

11. Method of manufacturing an analysis card (1) according to any one of the preceding claims, wherein the heating block comprises a second part of the heating element whose shape is complementary to that of the first face (10a) of the plate (9), the welding of the two films (3) of the receiving space (4) to the plate (9) comprises the welding of a first film (3a) to the first face (10a), the first film (3a) being pressed onto the first face (10a).

12. Analysis card (1) manufactured by the process according to any one of claims 1 to 11.

13. Heating block (14) configured to implement a method according to any one of claims 1 to 11, said heating block (14) comprising a support (15) and a heating element (16), a first part of the heating element (160) being configured to protrude at least during the application of the heating block (14), said first part of the heating element (160) then having a profile complementary to at least a part of the profile of the edge (11) of the plate 0).

14. Heating block (14) according to the preceding claim, wherein the first part of the heating element (160) comprises non-heating zones (26) whose width is greater than the width of the channels (13).

15. Heating block (14) according to the preceding claim, in which the non-heating areas (26) of the first part of the heating element (160) are recesses (26).

16. Heating block (14) according to any one of the preceding claims, wherein the first part of the heating element (160) comprises flexible nichrome (NiCr) elements.

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