Support for microscopic analysis of a liquid-based biological substance and system comprising such a support and a microscope

Abstract

The support comprises a slide (11) and a coverslip (12) secured by lines of glue (13) peripherally delimiting individual compartments (14) for receiving a sample of said biological substance; characterized in that the upper surface (17) of the slide is bare and exposed over at least one anterior marginal region (41) and at least one posterior marginal region (42), each having an extension in the anterior-posterior direction of at least 2.5 mm and an extension in the left-to-right direction of at least equal to the distance between the respective axes of the two most distant individual compartments. The system comprises the support (40) and a microscope configured to clamp the support between a stage and a support plate provided with fingers whose distal ends are in contact exclusively with the marginal regions (41, 42). (Figure 2)

Description

Title of the invention: Support for the microscopic analysis of a liquid-based biological substance and a system comprising such a support and a microscope Technical field of the invention

[0001] The invention relates to the microscopic analysis of a liquid-based biological substance, such as animal semen. More particularly, the invention relates to a support for such a substance, intended to be mounted in a microscope configured to perform such an analysis, as well as a system comprising such a support and such a microscope. State of the art

[0002] A support for the microscopic analysis of a liquid-based biological substance, such as animal semen, comprising a slide, a coverslip, and lines of glue arranged between the slide and the coverslip to fix them to each other while peripherally delimiting individual compartments for receiving a sample of said biological substance;the slide holder being parallelepiped and having two principal faces, namely a top face and a bottom face, and lateral faces extending from one to the other of the principal faces, namely a left face, a right face, a front face and a back face, with the distance between the left face and the right face being greater than the distance between the front face and the back face which is itself greater than the distance between the bottom face and the top face; said holder having a left-right direction transverse to the left face and the right face while it is parallel to the front face, the back face and the principal faces, a front-back direction transverse to the front face and the back face while it is parallel to the left face, the right face and the principal faces, and a bottom-up direction transverse to the principal faces while it is parallel to the lateral faces;the coverslip being parallelepiped and having two principal faces, namely a superior face and a inferior face, and lateral faces which extend from one to the other of the principal faces, namely a left face, a right face, a front face and a rear face; the inferior face of the coverslip being opposite the superior face of the slide while the front face, the rear face, the left face and the right face of the coverslip are turned towards and at a distance from respectively the front face, the rear face, the; left face and right face of the slide; each individual compartment being delimited in the bottom-up direction respectively by the upper face of the slide and by the lower face of the coverslip and peripherally delimited by the glue lines, each individual compartment opening exclusively on one side into a well and on the other side into a vent, each well and each vent being delimited by the glue lines, each well being turned towards one of the front and rear faces of the slide and each vent being turned towards the other of the front and rear faces of the slide.

[0003] To analyze the biological substance, a drop is placed in each of the wells of the support. The drops are then drawn by capillary action into the individual compartment with which the well communicates. The support is mounted in a microscope, and the individual compartments are arranged one after the other under its objective lens to observe the substance.

[0004] A system comprising such a support and such a microscope is also known, including an objective lens, a stage on which the support rests with the underside of the slide against the stage, and one of the individual compartments aligned with the objective lens. The microscope further comprises a frame on which the objective lens is mounted and on which the stage is mounted to move in translation along the left-right direction of the support so as to allow successive alignment of the other individual compartments with the objective lens. The microscope further comprises a support plate and a clamping element configured to bring the support plate and the stage towards each other. The support plate is hinged to the stage so that it can be moved away from the support or folded down onto the support. The system is further configured to focus the objective lens by autofocus. Description of the invention

[0005] The invention proposes, in a first aspect, a support similar to that described above but with improved performance, in particular with regard to the positioning of the individual compartments in relation to the objective of the microscope.

[0006] The invention provides for this purpose a support for the microscopic analysis of a liquid-based biological substance, such as animal semen, comprising a slide, a coverslip, and lines of adhesive arranged between the slide and the coverslip to secure them to one another while peripherally delimiting individual compartments for receiving a sample of said biological substance; the slide being parallelepiped-shaped and having two principal faces, namely a superior face and a inferior face, and lateral faces extending from one to the other of the principal faces, namely a left face, a right face, a front face, and a rear face, with the distance between the left face and the right face which is greater than the distance between the front face and the rear face which is itself greater than the distance between the lower face and the upper face; said support having a left-right direction transverse to the left face and the right face while it is parallel to the front face, the rear face and the principal faces, a front-back direction transverse to the front face and the rear face while it is parallel to the left face, the right face and the principal faces, and a bottom-up direction transverse to the principal faces while it is parallel to the lateral faces; the coverslip being parallelepiped-shaped and having two principal faces, namely a top face and a bottom face, and lateral faces which extend from one to the other of the principal faces, namely a left face, a right face, a front face and a rear face;the lower face of the coverslip being opposite the upper face of the slide, while the front, rear, left and right faces of the coverslip are turned towards and at a distance from respectively the front, rear, left and right faces of the slide; each said individual compartment being delimited in the bottom-up direction respectively by the upper face of the slide and by the lower face of the coverslip and delimited peripherally by said glue lines, each said individual compartment opening exclusively on one side onto a well and on the other side onto a vent, each well and each vent being delimited by said glue lines, each well being turned towards one of the front and rear faces of the slide and each vent being turned towards the other of the front and rear faces of the slide;characterized in that the upper face of the slide is bare and exposed on at least one front marginal region and on at least one rear marginal region, the front marginal region running along the front face of the slide, the rear marginal region running along the rear face of the slide, each of said front marginal region and rear marginal region having an extension along the front-to-back direction of at least 2.5 mm and an extension along the left-to-right direction of at least equal to the distance between the respective axes of the two individual compartments furthest from each other and at most equal to the distance between the left face and the right face of the slide.

[0007] Thanks to the marginal regions, it is possible to position the support according to the invention in relation to a microscope by pressing on the upper face of the slide holder, and not on its lower face as is conventionally done.

[0008] This allows for particularly precise positioning along the bottom-to-top direction of the sample to be analyzed, since it avoids uncertainty on the thickness of the specimen holder due to manufacturing tolerances.

[0009] It should be noted that the expression "bare" refers to the upper surface of the slide. means that this upper surface does not directly bear any locally added coating, such as lines of glue or ink.

[0010] It should also be noted that the expression "uncovered" referring to the upper face of the slide means that this upper face is not in view of any element (such as the coverslip) which could constitute an obstacle for another element which would be brought closer to the support in the bottom-up direction to come into contact with its upper face.

[0011] Thanks to the marginal regions, it is possible to directly access the upper surface of the slide in order to mechanically determine its position and thus determine the position of other elements whose position is fixed and / or known relative to this upper surface. It should be noted in particular that mechanically determining the position of the upper surface is equivalent to mechanically determining the position of the individual compartments that the upper surface delimits.

[0012] The invention is further based on the observation that the optical quality of the surface of the specimen holder slide (necessary in order to disturb as little as possible the light which passes through the slide to illuminate the sample present in the individual compartment) implies a mechanical regularity of this surface which is favorable to sliding; and that it is in particular possible to exert pressure on this surface while retaining a sliding capacity on this surface.

[0013] The front and rear marginal regions can thus serve as sliding tracks for at least one support element - such as, for example, a finger as explained later - against which the upper face would be supported by its marginal regions in order to have its position mechanically determined.

[0014] It should also be noted that the position of the marginal regions - namely along the front face and the rear face - means that these regions are as far apart from each other as possible, which is favorable to the accuracy of the mechanical determination of the positioning by the support elements and to stability during sliding.

[0015] With regard to the lower limit of the extension of the marginal regions along the front-back direction - namely 2.5 mm -, the applicant determined that, taking into account the manufacturing tolerances of the various elements of the system, a value of at least 2.5 mm made it possible to guarantee that the support fingers remain exclusively in contact with the front and rear marginal regions, regardless of the position of the plate along the left-right direction.

[0016] Particularly simple, convenient and economical advantageous features of the support according to the invention are presented below: - for at least one of the anterior marginal region and the rear marginal region, the extension along the left-right direction is equal to the distance between the left face and the right face of the slide; - each well has an extension along the front-to-back direction which is less than the extension along the front-to-back direction of each of said front marginal region and rear marginal region; - the distance between the left and right faces of the microscope slide is between 74.8 mm and 75.2 mm, the distance between the front and back faces of the microscope slide is between 24.8 mm and 25.2 mm, the distance between the left and right faces of the coverslip is between 31.8 mm and 32.2 mm, and the distance between the front and back faces of the coverslip is between 16.8 mm and 16.2 mm; and / or - all the wells are turned towards one of the front and rear faces of the slide holder and all the vents are turned towards the other of the front and rear faces of the slide holder. The invention proposes, in a second aspect, a system for the analysis of a liquid-based biological substance, comprising a support as described above and a microscope comprising an objective, a stage configured to receive the support in a first observation position in which the lower face of the slide holder is against the stage and one of the individual compartments is aligned with the objective, a frame on which the objective is mounted in at least one fixed predetermined position and on which the stage is mounted movable in translation along the left-right direction of the support in order to successively align the other individual compartments with the objective, a support plate extending at least partially opposite the support, and a clamping member configured to stress the support plate and the stage towards each other so that the support is clamped between the support plate and the stage;the support plate being fixedly mounted on the frame and comprising a base and support fingers projecting from the base towards the plate and whose distal ends are in contact exclusively with the marginal regions, with at least two support fingers on one of the front marginal region and rear marginal region and at least one support finger on the other of the front marginal region and rear marginal region; the plate having positioning stops to immobilize the support relative to the plate in the left-right direction.

[0017] Since the support plate is fixedly mounted on the frame and the upper face of the slide holder is held against the support fingers by the plate, under the effect of the clamping member, the support fingers determine the position of the upper face relative to the frame in the bottom-up direction.

[0018] It should be noted that in this way the position of the upper face relative to the frame does not depend of the thickness (i.e. the distance between the lower face and the upper face) of the slide.

[0019] During the translation of the plate in the left-right direction, the fingers The support points slide on the marginal regions. Since the left-to-right extension of the marginal regions is at least equal to the distance between the respective axes of the two most distant individual compartments, the fingers remain in contact with the marginal regions regardless of which individual compartment is aligned with the objective.

[0020] Thus, the position of the upper face of the slide holder along the bottom-up direction remains determined in the same way by the support fingers regardless of the individual compartment aligned with the objective.

[0021] This allows, in particular, once the focus of the objective has been made for a given individual compartment, to maintain this focus for each of the other individual compartments.

[0022] This also allows this focus to be maintained for another support put in place to replace a first support for which the focus has been made, since the position following the bottom-up direction of the upper face of this other support will be determined by the support fingers in the same way as for the first support.

[0023] It should be noted that such a setting may then correspond to a factory setting made during the manufacture of the microscope.

[0024] It should also be noted that in prior art systems, the mechanical reference surface is the surface of the stage on which the lower face of the slide rests, so that the position of the upper face of the slide depends on the thickness of the slide, and that consequently these systems do not offer the advantages set out above.

[0025] Particularly simple, convenient and economical advantageous features of the system according to the invention are presented below: - the support plate has four support fingers, two of which are in contact with the front marginal region and the other two are in contact with the rear marginal region; - One part of the mounting plate and support plate has magnetic properties, and the clamping element includes magnets attached to the other part of the mounting plate and support plate. - the base has an opening through which the lens points to said individual compartment aligned with the lens in said first observation position, at least two of the support fingers which are in contact with the same marginal region being arranged on either side of the opening; - the frame comprises a mounting wall and the base comprises two arms, each projecting transversely from the mounting wall, extending opposite each other and defining the opening, each arm being provided with two support fingers, of which one finger in contact with the anterior marginal region and one finger in contact with the posterior marginal region; - the clamping device includes magnets housed in said arms; - one of the plate and the support plate has a ramp and the other of the plate and the support plate has a boss configured to cooperate with the ramp during the translational movement of the plate along the left-right direction so as to move the plate away from the support plate, against the force exerted by the clamping member, to a position in which the support is accessible so that it can be removed from the plate; - the plate is mechanically connected to the frame by a sliding pivot joint comprising a cylindrical guide longitudinally oriented in the left-right direction and a shaft mounted to slide on the guide, with a play between the shaft and the guide; and / or - the lens has a fixed focal length.

[0026] The invention also relates to the use of a system as described above in which the liquid-based biological substance is animal semen and said analysis includes a sperm counting step. Brief description of the figures

[0027] We will now continue the exposition of the invention with a detailed description of exemplary embodiments, given below by way of illustration and not limitation, with reference to the accompanying drawings. These drawings: - Fig. 1 is a view of a support for the microscopic analysis of a liquid-based biological substance, in accordance with the prior art. - Fig. 2 is a view similar to Fig. 1, and at the same scale, of a support according to the invention. - Fig. 3 is a very schematic cross-sectional view of a system according to the invention (illustrated in more detail in Figures 9 to 12), comprising the support shown in Fig. 2 and a microscope, the system being in an observation configuration in which the support is clamped between a stage and the fingers of a microscope support plate with one of the individual compartments of the support aligned with the objective; the cross-section being along the front-back direction of the support and passing through the middle of this individual compartment. - Fig. 4 is a cross-sectional view of the system illustrated in Fig. 3, this time following the left-right direction of the support and passing through the middle of the individual compartments. - Fig. 5 is a view similar to Fig. 4 after the plate has been moved in the left-right direction to align another of the individual compartments with the objective. - Figure 6 is a view similar to Figure 2, showing the axes of the individual compartments, as well as the positions of the fingers of the support plate on the upper surface of the slide when the stage is in the position shown in Figure 4 and when the stage is in the position shown in Figure 5. - Figure 7 is a view similar to Figure 6, showing the path of each of the fingers of the support plate when the stage is moved from the position shown in Figure 4 to the position shown in Figure 5. - Fig. 8 is a view similar to Fig. 3, after the turntable has been moved away from the support plate to move the system into a loading / unloading configuration in which the support is accessible to a system user. - Fig. 9 is an exploded and detailed perspective view of the system schematically illustrated in figures 2 to 8, but without the lens. - Fig. 10 is a view similar to Fig. 9 but for the system in the assembled state, and in a cleaning configuration in which the turntable is very far from the support plate. - Fig. 11 is a cross-sectional view of the system as illustrated in Figures 9 and 10, taken along the front-to-back direction of the support and passing through one of the individual compartments, the system being in the loading / unloading configuration schematically represented in Fig. 8. - Fig. 12 is a side view of part of the system illustrated in Figures 9 to 11, the system being in the observation configuration schematically represented in Figures 3 to 5. - Fig. 13 is a view similar to Fig. 7 for a variant of the support according to the invention. Detailed description

[0028] The support 10 and the support 40 illustrated respectively in figures 1 and 2 each comprise a specimen slide 11, a coverslip 12 and lines of glue 13 arranged between the specimen slide 11 and the coverslip 12 to fix them to each other while peripherally delimiting individual compartments 14.

[0029] It should be noted beforehand that in the present report, the same numerical references have been used for the corresponding elements of support 10 and support 40 for simplicity.

[0030] Therefore, in the description of the support 10 that follows, if reference is made to an element of the support 10 which is not visible on [fig.1], it is possible - for the purpose of understanding - to refer to one of the figures illustrating the support 40 on which the corresponding element is visible.

[0031] The individual compartments 14 of the support 10 are each configured to receive a sample of the liquid-based biological substance, here animal semen, diluted or not.

[0032] The support 10 here comprises individual compartments 14.

[0033] Each individual compartment 14 opens exclusively on one side onto a well 15 and on the other side onto a vent 16, each well 15 and each vent 16 being delimited by the glue lines 13.

[0034] To fill an individual compartment 14 with a sample of the biological substance, a drop (not shown) of this substance is deposited in the respective well 15. This drop is then drawn by capillary action into the individual compartment 14 while the air initially present in the individual compartment 14 escapes through the vent 16.

[0035] The specimen holder 11 and the specimen coverslip 12 are here each made of float glass coated with a surfactant film promoting the entrainment of the drop into the individual compartment 14 by capillarity.

[0036] The specimen holder slide 11 and the specimen coverslip 12 are each transparent.

[0037] The slide holder 11 is parallelepiped-shaped and has two main faces, at namely an upper face 17 and a lower face 18 (figures 3 and 4), and lateral faces extending from one to the other of the main faces, namely a left face 19, a right face 20, a front face 21 and a rear face 22.

[0038] The distance between the left face 19 and the right face 20 is greater than the distance between the front face 21 and the rear face 22, which is itself greater than the distance between the lower face 18 and the upper face 19.

[0039] The distance between the left face 19 and the right face 20 is here between 74.8 mm and 75.2 mm.

[0040] The distance between the front face 21 and the rear face 22 is here between 24.8 mm and 25.2 mm.

[0041] The distance between the lower face 18 and the upper face 19 - i.e. the thickness of the slide 11 - is here about 1 mm.

[0042] The support 10 has a left-right direction transverse to the left face 19 and the right face 20 while it is parallel to the front face 21, the rear face 22 and the main faces 17 and 18, a front-back direction transverse to the front face 21 and the rear face 22 while it is parallel to the left face 19, the right face 20 and the main faces 17 and 18, and a bottom-up direction transverse to the main faces 17 and 18 while it is parallel to the lateral faces 19, 20, 21 and 22.

[0043] The coverslip 12 is parallelepiped-shaped and has two main faces, namely a superior face 23 and a inferior face 24, and lateral faces extending from one to the other of the main faces, namely a left face 25, a right face 26, a front face 27 and a rear face 28.

[0044] The distance between the left face 25 and the right face 26 is greater than the distance between the front face 27 and the rear face 28, which is itself greater than the distance between the lower face 24 and the upper face 23.

[0045] The distance between the left face 25 and the right face 26 is here between 31.8 mm and 32.2 mm.

[0046] The distance between the front face 27 and the rear face 28 is here between 20.8 mm and 21.2 mm.

[0047] The distance between the lower face 24 and the upper face 23 - i.e. the thickness of the coverslip 12 - is here 0.7 mm + / - 0.05 mm.

[0048] The lower face 24 of the coverslip 12 is opposite the upper face 17 of the slide 11 while the front face 27, the rear face 28, the left face 25 and the right face 26 of the coverslip 12 are turned towards and at a distance from respectively the front face 21, the rear face 22, the left face 19 and the right face 20 of the slide 11.

[0049] The coverslip 12 is here arranged centered relative to the slide 11 in the left-right direction.

[0050] Each individual compartment 14 is delimited in the bottom-up direction respectively by the upper face 17 of the specimen holder slide 11 and by the lower face 24 of the specimen coverslip 12 and delimited peripherally by the glue lines 13.

[0051] The thickness of the glue lines 13 is here 20 µm.

[0052] It should be noted that in figures 3 to 5 and 8, the thickness of the glue lines 13 has been exaggerated for the sake of readability.

[0053] Each individual compartment 14 here has a capacity of 3 ft.

[0054] The individual compartments 14 follow one another in the left-right direction, here being juxtaposed one to the other.

[0055] As explained above, each individual compartment 14 opens on one side onto a well 15 and on the other side onto a vent 16, the well 15 being here turned towards the rear face 22 of the slide holder 11 and the vent 16 being here turned towards the front face 21 of the slide holder 11.

[0056] Here all the wells 15 are turned towards the rear face 22 and all the vents 16 are turned towards the front face 21.

[0057] Each well 15 is here delimited by a portion 29 of the glue lines 13 shaped in a U and extending beyond the space strictly between the slide 11 and the coverslip 12 to an end 30 located here flush with the rear face 22 of the slide 11.

[0058] Each vent 16 is here delimited by straight portions 31 of the glue lines 13 extending opposite each other, each overhanging the strictly defined space included between the specimen slide 11 and the coverslip 12 and up to a respective end 32 located here flush with the front face 21 of the specimen slide 11.

[0059] The support 40 illustrated in [fig.2] is similar to the support 10, except that the respective dimensions of the coverslip 12 and the glue lines 13 of the support 40 are smaller in the front-back direction and that the upper face 17 of the slide 11 of the support 40 is bare and exposed on a front marginal region 41 and on a rear marginal region 42.

[0060] More specifically, the distance between the front face 27 and the rear face 28 of the coverslip 12 is here between 16.8 mm and 16.2 mm.

[0061] It will be noted that the length of the individual compartments 14 of the support 40, that is to say the distance separating the well 15 from the vent 16, which here corresponds to the distance between the front face 27 and the rear face 28 of the coverslip 12, is therefore also smaller than that of the individual compartments 14 of the support 10.

[0062] The front marginal region 41 runs along the front face 21 of the slide holder 11, while the rear marginal region 42 runs along the rear face 22 of the slide holder 11.

[0063] Here, the front marginal region 41 and the rear marginal region 42 are each directly bordered by the front face 21 and the rear face 22 of the specimen holder 11 respectively.

[0064] The front marginal region 41 here has an extension along the front-back direction equal to 2.5 mm and has an extension along the left-right direction equal to the distance between the left face 19 and the right face 20 of the slide holder 11.

[0065] The limit of the extension along the front-back direction of the front marginal region 41 is located on the [fig.2] by the broken line 43, where it passes close to each of the ends 32 of the vents 16.

[0066] Indeed, beyond this limit 43 going towards the rear, the upper face 17 is covered by the glue lines 13 and / or is opposite the lower face 24 of the coverslip 12 and is therefore not bare and exposed.

[0067] The rear marginal region 42 here has an extension along the front-to-back direction equal to 2.6 mm and has an extension along the left-to-right direction equal to the distance between the left face 19 and the right face 20 of the slide holder 11.

[0068] The limit of the extension along the front-back direction of the rear marginal region 42 is located on the [fig.2] by the broken line 44, where it passes close to the ends 30 of the wells 15.

[0069] Indeed, beyond this limit 44 going forward, the upper face 17 is covered by the glue lines 13 and / or is opposite the lower face 24 of the coverslip 12 and is therefore not bare and exposed.

[0070] Generally speaking, each of the marginal regions before 41 and marginal region rear 42 has an extension along the front-to-rear direction of at most equal to 3 mm, or alternatively equal to more than 3 mm, for example 3.5 mm, 4 mm, 4.5 mm, 5 mm, or even more than 5 mm.

[0071] In addition, it will be noted that the extension along the front-back direction of each of the wells 15 - that is to say the distance, measured at the level of the upper face 17, between the rear face 28 of the coverslip 12 and the limit 44 (which passes flush with the ends 30 of the wells 15) - is here equal to 2.4 mm, which is notably less than both the extension along the front-back direction of the front marginal region 41 (2.5 mm) and the extension along the front-back direction of the rear marginal region 42 (2.6 mm).

[0072] Figures 3 to 5 and 8 illustrate a system 100 for the analysis of the liquid-based biological substance, comprising the support 40 and a microscope 50.

[0073] In figures 3 to 5, the system is in an observation configuration in which the microscope 50 is cooperating with the support 40 in order to observe the samples of biological substance contained in the individual compartments 14.

[0074] The microscope 50 includes an objective 51 and a stage 52 configured to receive the support 40 in at least one observation position in which the lower face 18 of the slide holder 11 is against the stage 52 and one of the individual compartments 14 is aligned with the objective 51.

[0075] In figures 3 and 4, the support 40 is in a first observation position in which the compartment 14 aligned with the objective 51 is the one that can be seen furthest to the left in [fig.2].

[0076] As explained in more detail below, the microscope 50 is configured to dispose of the support 40 in a plurality of observation positions in each of which the stage 52 receives the support 40 with a respective individual compartment 14 aligned with the objective 51.

[0077] On [fig.5] for example, the support 40 is in a second observation position in which the compartment 14 aligned with the objective 51 is the one that is seen furthest to the right on [fig.2].

[0078] The microscope 50 further comprises a frame (not illustrated in Figures 3 to 5 and 8 but of which a mounting wall 60 is visible in Figures 9 to 11) on which the objective 51 is fixedly mounted and on which the stage 52 is movable in translation along the left-right direction of the support 40 in order to be able to successively align the other individual compartments 14 with the objective 51.

[0079] The mechanical connection between the plate 52 and the frame will be described in detail later, with reference to Figures 9 to 12.

[0080] The plate 52 is further configured to receive the support 40 in a position predetermined.

[0081] To position and immobilize the support 40 in this predetermined position, the plate 52 here includes positioning stops each configured to cooperate with a respective lateral face of the object-carrying slide 11 with which it is opposite in order to immobilize the support 40 in the direction (left-right or front-back as the case may be) transverse to this respective lateral face.

[0082] The positioning stops here comprise two front positioning stops 33 opposite the front side face 21, a left positioning stop 34 opposite the left side face 19, a right positioning stop 35 opposite the right side face 20 and two rear positioning stops 36 opposite the rear side face 22, so that the support 40 is here immobilized relative to the plate 52 both in the front-to-back direction in both directions and in the left-to-right direction in both directions.

[0083] The positioning stops are here formed by pins protruding from an upper face 37 of the plate 52 on which rests the lower face 18 of the slide holder 11.

[0084] The plate 52 further includes a notch 45 which extends opposite each of the individual compartments 14 and is configured to allow light from a lighting element (not shown) disposed under the plate 52 to reach each of the individual compartments 14.

[0085] The microscope 50 further includes a support plate 53 which extends at least partially in relation to the support 40, as well as a clamping member configured to stress the support plate 53 and the stage 52 towards each other, so that the support 40 is clamped between the support plate 53 and the stage 52.

[0086] The clamping member here includes magnets 54 attached to the support plate 53, while the plate 52 has magnetic properties, the magnets 54 being configured to strain the plate 52 towards the support plate 53.

[0087] The support plate 53 is fixedly mounted on the frame and includes a base 55 and support fingers 56 each projecting from the base 55 towards the plate 52 to a distal end 57.

[0088] The distal ends 57 of the support fingers 56 are each exclusively in contact with the marginal regions 41 and 42 of the upper face 17 of the specimen-bearing blade 11.

[0089] The slide holder 11 is thus held between the support fingers 56 and the plate 52, the slide holder 11 being in contact with the support plate 53 here exclusively through the support fingers 56.

[0090] The support plate 53 here comprises four fingers 56, two of which are in contact with the marginal region before 41 and two other fingers 56 are in contact with the rear marginal region 42.

[0091] The four supporting fingers 56 are arranged here so that each is located at the respective vertex of an imaginary rectangle.

[0092] The base 55 further has an aperture 58 through which the lens 51 points to the individual compartment 14 aligned with the lens 51, the latter being here partially received in the aperture 58.

[0093] The support fingers 56 which are in contact with the front marginal region 41 are arranged on either side of the opening 58. The support fingers 56 which are in contact with the rear marginal region 42 are arranged on either side of the opening 58.

[0094] The lens 51 here has a fixed focal length.

[0095] The system 100 is configured so that the focal point 59 of the lens 51 is located here approximately in the middle of the individual compartment 14 which is aligned with the lens 51.

[0096] During the translation of the plate 52 along the left-right direction of the support 40, the support fingers 56 slide on the respective marginal regions 41 and 42, each remaining exclusively in contact with them.

[0097] On [fig.6], the axes 38 of each of the individual compartments 14 have been identified.

[0098] When an individual compartment 14 is aligned with the objective 51 to observe the sample received in this compartment 14, the axis 38 of this compartment 14 is more precisely aligned with the optical axis of the objective 51.

[0099] Consequently, between the position it occupies on [fig.4] and the position it occupies on [fig.5], the plate 52 has been moved over a distance equal to the gap between the respective axes 38 of the leftmost individual compartment 14 and the rightmost individual compartment 14, i.e. the two individual compartments 14 furthest from each other on the support 40.

[0100] The gap between the axes 38 of two neighboring compartments is here equal to 7.8 mm and the gap between the axes 38 of the two individual compartments 14 furthest from each other is therefore 23.4 mm.

[0101] On [fig.6], the respective positions of the support fingers 56 when the plate 52 is in the position it occupies on [fig.4] have also been identified by four black rectangles 46, and the respective positions of these fingers 56 when the plate 52 is in the position it occupies on [fig.5] have been identified by four hatched rectangles 47.

[0102] Each black rectangle 46 or hatched rectangle 47 corresponds to the portion of the upper face 17 which is in contact with the distal end 57 of a support finger 56, this portion having here an extension along the front-back direction equal to about 1 mm and an extension along the left-right direction equal to about 3 mm.

[0103] In [Fig. 7], the path of each of the support fingers 56 when the plate 52 moves from the position it occupies on the [Fig. 7] has also been marked by hatched rectangles 48. [fig.4] to the position it occupies on the [fig.5].

[0104] Each hatched rectangle 48 therefore corresponds to the integral of the respective rectangle 46 or 47 over the entire path of the corresponding finger 56. Each rectangle 48 thus has an extension in the front-back direction equal to approximately 1 mm and an extension in the left-right direction equal to approximately 3 mm plus the length of the path of the finger 56, this latter length being - as explained above - equal to the distance between the respective axes 38 of the leftmost individual compartment 14 and the rightmost individual compartment 14, that is to say the two individual compartments 14 furthest from each other on the support 40.

[0105] It will be noted that in order for each support finger 56 to remain entirely in contact with the respective marginal region 41 or 42 which it travels through, regardless of which individual compartment 14 is aligned with the objective 51, the extension along the left-right direction of this marginal region must be at least equal to the extension along the left-right direction of the hatched rectangles 48.

[0106] It will also be noted that the extension along the left-right direction of the hatched rectangles 48 is in practice greater than the distance between the respective axes 38 of the two individual compartments 14 furthest from each other on the support 40 (since it is necessary to take into account the extension of the support fingers along the left-right direction).

[0107] It should also be noted that in [Fig. 7], the hatched rectangles 48 of the same marginal region 41 or 42 do not overlap, so that a coating could be present between these two rectangles 48 without being encountered by a support finger 56 during its path. Thus, in an unillustrated variant, the support has two front marginal regions and / or two rear marginal regions as described above, but each with a smaller extension in the left-right direction, and with a coating interposed between these two front marginal regions and / or between these two rear marginal regions.

[0108] The support 70 illustrated in [fig.13] is similar to the support 40 except that it only has two individual compartments 14, the distance between the axes 38 of these compartments being in this case equal to 15.84 mm.

[0109] Fig. 8 illustrates a loading / unloading configuration of the system 100, here obtained after the turntable 52 has been moved away from the support plate 53, the support 40 then being accessible to a user to be removed from the turntable 52 and possibly replaced by another similar support in order to analyze the samples contained in the compartments of this other support.

[0110] Conversely, to switch the system 100 from the loading / unloading configuration to the observation configuration, the platform 52 loaded with the support 40 is brought closer to the support plate 53 until each support finger 56 is at contact of marginal regions 41 and 42 in the manner described previously.

[0111] We will now describe system 100 in more detail with reference to Figures 9 to 12.

[0112] The frame here comprises a mounting wall 60 and the base 55 comprises two branches 61 each projecting transversely from the mounting wall 60. The branches 61 extend opposite each other, delimiting the opening 58.

[0113] Each branch 61 is here provided with two support fingers 56, of which one finger 56 is in contact with the front marginal region 41 and one finger is in contact with the rear marginal region 42 when the system 100 is in the observation configuration ( [fig-12]).

[0114] As can be clearly seen in [Fig. 10], the magnets 54 of the clamping member are housed in the arms 61, each arm 61 being provided with three cylindrical magnets 54. For each arm 61, the magnets 54 are distributed between the support fingers 56 of that arm 61.

[0115] The support plate 53 here has guide grooves 62 provided in the base 55, on the same side as the support fingers 56, each guide groove 62 being configured to receive an end portion of a respective pin forming one of the front positioning stops 33, left 34 and / or right 35 of the plate 52 when the system 100 is in observation configuration, the ends of the pins moving in the respective guide groove when the plate 52 moves in the left-right direction.

[0116] The guide grooves 62 thus contribute to the correct positioning of the plate 52 along the front-back direction of the support 40, that is to say so that the support fingers 56 remain exclusively in contact with the marginal regions 41 and 42 during the movement of the plate 52.

[0117] The plate 52 further has a ramp 63 and the support plate 53 has a boss 64 configured to cooperate with the ramp 63 during the translational movement of the plate 52 along the left-right direction of the support 40 so as to move the plate 52 away from the support plate 53, against the force exerted by the clamping member, here until the plate arrives in the position illustrated in [Fig. 11], where the system 100 is in its loading / unloading configuration.

[0118] The plate 52 is here mechanically connected to the frame by a sliding pivot joint comprising a cylindrical guide 65 and a shaft 66 mounted to slide on the guide 65, the latter being longitudinally oriented in a direction parallel to the left-right direction of the support 40 when the latter is received in its predetermined position on the plate 52.

[0119] Such a mechanical link makes it possible both to move the plate 52 away from and closer to the support plate 53 and to move the plate 52 in the left-right direction.

[0120] The guide 65 is here fixed to the mounting wall 60 while the barrel 66 is integral with turntable 52.

[0121] The internal diameter of the barrel 66 is here slightly greater than the external diameter of the guide 65 so that there is a play between them allowing the plate 52 to pivot to a certain extent both in the left-right direction in both directions and in the front-back direction in both directions.

[0122] This ability of the plate 52 to pivot allows the inclination of the upper face 17 of the slide holder 11 to be adjusted relative to the support fingers 56 in order to ensure that the latter are all in contact with the upper face 17, with support evenly distributed between all the fingers 56.

[0123] To move the stage 52 in the left-right direction, the microscope 50 further includes a motorized mechanism (not shown) controlled from a user interface. Alternatively, the mechanism is not motorized and is operated manually using a knob.

[0124] The system 100 as described above is particularly suitable for an analysis comprising a step of counting the spermatozoa present in animal semen samples, in order to determine their concentration.

[0125] In an unillustrated variant, the system 100 is coupled to a computer configured to automatically analyze the images captured by the lens in order to perform the counting step and determine the concentration.

[0126] In an unillustrated variant, the focal point of the lens is not in the middle of the individual compartments but is located - along the bottom-up direction - at a predetermined distance from the middle of the individual compartments.

[0127] In an unillustrated variant, the objective is mounted movable relative to the microscope frame between a plurality of fixed predetermined positions in each of which the objective is at a different respective distance from the support.

[0128] It is understood that in the present application, the term "objective" should be interpreted broadly and may include a simple electronic image sensor, such as a CMOS sensor, used for example in so-called "lensless" microscopy processes, employing, for example, the principle of online holography. Thus, alternatively, the analysis system as described above is provided with a camera equipped with such an electronic sensor and is configured to record and process a sequence of images in order to analyze the biological substance, for example, to determine the motility of spermatozoa contained in animal semen samples.

[0129] In another unillustrated embodiment, the front marginal strip and the rear marginal strip are located a short distance from the front face 21 or the rear face 22 respectively, i.e. for example, a coating or a portion of a coating is located on the upper face 17 between the front face 21 and the front marginal region and / or between the rear face 22 and the rear marginal region.

[0130] In yet another variant not illustrated, the support plate has three support fingers, with - when the system is in holding configuration - two of the fingers being exclusively in contact with one of the front and rear marginal regions and the last of the fingers being exclusively in contact with the other of the front and rear marginal regions.

[0131] In yet another variant not illustrated, the clamping member includes magnets which are fixed to the plate while the support plate has magnetic properties.

[0132] In yet another variant not illustrated, the positioning stops are different from protruding pins on the upper face 37 of the plate 52 and are for example formed by the edges of a recess made in this upper face 37 and configured to accommodate the specimen holder 11.

[0133] In still other variants not illustrated, the wells and vents are arranged differently than described above, for example all the wells are turned towards the front face and all the vents are turned towards the rear face, or from one compartment to another the wells and vents are alternately turned towards the front face and then towards the rear face.

[0134] In other variants not illustrated: - the lens has a variable focal length rather than a fixed one; - the number of individual compartments is different from four and two, for example three, five or more than five; - the clamping element has a number of magnets other than three per arm, for example one, two, four or more than four magnets per arm and / or the magnets have a shape other than cylindrical, for example parallelepiped; - the clamping element is different from magnets housed in the arms of the support plate, and includes, for example, springs arranged on the side of the plate facing away from the support plate and configured to push the plate towards the support plate; and / or - The ramp 63 is supported by the support plate and the boss 64 is supported by the base plate.

[0135] Many other variations are possible depending on the circumstances, and it should be noted in this regard that the invention is not limited to the examples described and illustrated.

Claims

Demands

1. Support for the microscopic analysis of a liquid-based biological substance, such as animal semen, comprising a slide holder (11), a coverslip (12) and lines of glue (13) arranged between the slide holder (11) and the coverslip (12) to fix them to each other while peripherally delimiting individual compartments (14) for receiving a sample of said biological substance;the specimen holder (11) being parallelepiped and having two main faces, namely a top face (17) and a bottom face (18), and lateral faces which extend from one to the other of the main faces, namely a left face (19), a right face (20), a front face (21) and a back face (22), with the distance between the left face (19) and the right face (20) which is greater than the distance between the front face (21) and the back face (22) which is itself greater than the distance between the bottom face (18) and the top face (17);said support (40) having a left-right direction transverse to the left face (19) and the right face (20) while it is parallel to the front face (21), the rear face (22) and the principal faces, a front-back direction transverse to the front face (21) and the rear face (22) while it is parallel to the left face (19), the right face (20) and the principal faces, and a bottom-up direction transverse to the principal faces while it is parallel to the lateral faces; the coverslip (12) being parallelepiped-shaped and having two principal faces, namely a top face (23) and a bottom face (24), and lateral faces extending from one to the other of the principal faces, namely a left face (25), a right face (26), a front face (27) and a rear face (28);the lower face (24) of the coverslip (12) being opposite the upper face (17) of the slide (11) while the front face (27), the rear face (28), the left face (25) and the right face (26) of the coverslip (12) are turned towards and at a distance from respectively the front face (21), the rear face (22), the left face (19) and the right face (20) of the slide (11); each said individual compartment (14) being delimited in the bottom-up direction respectively by the upper face (17) of the slide (11) and by the lower face (24) of the coverslip (12) and peripherally delimited by said glue lines (13), each said com-; individual compartment (14) opening exclusively on one side onto a well (15) and on the other side onto a vent (16), each well (15) and each vent (16) being delimited by said glue lines (13), each well (15) being turned towards one of the front face (21) and rear face (22) of the slide holder (11) and each vent (16) being turned towards the other of the front face (21) and rear face (22) of the slide holder (11);characterized in that the upper face (17) of the slide (11) is bare and exposed on at least one front marginal region (41) and on at least one rear marginal region (42), the front marginal region (41) running along the front face (21) of the slide (11), the rear marginal region (42) running along the rear face (22) of the slide (11), each of said front marginal region (41) and rear marginal region (42) having an extension along the front-to-back direction of at least 2.5 mm and an extension along the left-to-right direction of at least equal to the distance between the respective axes (38) of the two individual compartments (14) furthest from each other and at most equal to the distance between the left face (19) and the right face (20) of the slide.;

2. Support according to claim 1, characterized in that for at least one of the front marginal region (41) and the rear marginal region (42), the extension along the left-right direction is equal to the distance between the left face (19) and the right face (20) of the specimen holder (11).

3. Support according to any one of claims 1 or 2, characterized in that each well (15) has an extension along the front-back direction which is less than the extension along the front-back direction of each of said front marginal region (41) and rear marginal region (42).

4. Support according to any one of claims 1 to 3, characterized in that the distance between the left face (19) and the right face (20) of the slide holder (11) is between 74.8 mm and 75.2 mm, the distance between the front face (21) and the rear face (22) of the slide holder (11) is between 24.8 mm and 25.2 mm, the distance between the left face (25) and the right face (26) of the coverslip (12) is between 31.8 mm and 32.2 mm, and the distance between the front face (27) and the rear face (28) of the coverslip (12) is between 16.8 mm and 16.2 mm.

5. Support according to any one of claims 1 to 4, characterized in that all the wells (15) are turned towards one of the front face (21) and rear face (22) of the specimen holder slide (11) and all the vents (16) are turned towards the other of the front face (21) and rear face (22) of the specimen holder slide (11).

6. A system for the analysis of a liquid-based biological substance, comprising a support (40) according to any one of claims 1 to 5 and a microscope (50) comprising an objective (51), a stage (52) configured to receive the support (40) in a first observation position in which the lower face (18) of the slide (11) is against the stage (52) and one of the individual compartments (14) is aligned with the objective (51), a frame on which the objective (51) is mounted in at least one fixed predetermined position and on which the stage (52) is mounted movable in translation along the left-right direction of the support (40) so as to be able to successively align the other individual compartments (14) with the objective (51), a support plate (53) extending at least partially opposite the support (40),and a clamping member (54) configured to press the support plate (53) and the plate (52) towards each other so that the support (40) is clamped between the support plate (53) and the plate (52); the support plate (53) being fixedly mounted on the frame and comprising a base (55) and support fingers (56) projecting from the base (55) towards the plate (52) and whose distal ends (57) are in contact with the marginal regions (41, 42) exclusively, with at least two support fingers (56) on one of the front marginal region (41) and rear marginal region (42) and at least one support finger (56) on the other of the front marginal region (41) and rear marginal region (42); the plate (52) having positioning stops (34, 35) to immobilize the support (40) relative to the plate (52) in the left-right direction.

7. System according to claim 6, characterized in that the support plate (53) has four support fingers (56), of which two support fingers (56) are in contact with the front marginal region (41) and the other two support fingers (56) are in contact with the rear marginal region (42).

8. System according to any one of claims 6 or 7, characterized in that one of the plate (52) and the support plate (53) has magnetic properties and the clamping member comprises magnets (54) integral with the other of the plate (52) and the support plate (53).

9. A system according to any one of claims 6 to 8, characterized in that the base (55) has an opening (58) through which the objective (51) points to said individual compartment (14) aligned with the objective (51) in said first observation position, at least two of the support fingers (56) which are in contact with the same marginal region (41, 42) being arranged on either side of the opening (58).

10. System according to claim 9, characterized in that the frame comprises a mounting wall (60) and the base (55) comprises two arms (61) each projecting transversely from the mounting wall (60), extending opposite each other and delimiting the opening (58), each arm (61) being provided with two support fingers (56), one finger (56) in contact with the front marginal region (41) and one finger (56) in contact with the rear marginal region (42).

11. System according to claim 10, characterized in that the clamping member comprises magnets (54) housed in said arms (61).

12. System according to any one of claims 6 to 11, characterized in that one of the plate (52) and the support plate (53) has a ramp (63) and the other of the plate (52) and the support plate (53) has a boss (64) configured to cooperate with the ramp (63) during the translational movement of the plate (52) in the left-right direction so as to move the plate (52) away from the support plate (53), against the force exerted by the clamping member (54), to a position in which the support (40) is accessible so as to be removed from the plate (52).

13. System according to any one of claims 6 to 12, characterized in that the plate (52) is mechanically connected to the frame by a sliding pivot joint comprising a cylindrical guide (65) longitudinally oriented in the left-right direction and a shaft (66) mounted to slide on the guide (65), with a clearance between the shaft (66) and the guide (65).

14. System according to any one of claims 6 to 13, characterized in that the objective (51) has a fixed focal length.

15. Use of a system according to any one of claims 6 to 14, characterized in that the liquid-based biological substance is animal semen and said analysis includes a sperm counting step.

Images