EP4267302A1 - Support for analysing by microscope a liquid-based biological substance and system comprising such a support and a microscope - Google Patents

Support for analysing by microscope a liquid-based biological substance and system comprising such a support and a microscope

Info

Publication number
EP4267302A1
EP4267302A1 EP21851668.0A EP21851668A EP4267302A1 EP 4267302 A1 EP4267302 A1 EP 4267302A1 EP 21851668 A EP21851668 A EP 21851668A EP 4267302 A1 EP4267302 A1 EP 4267302A1
Authority
EP
European Patent Office
Prior art keywords
face
support
plate
slide
marginal region
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
EP21851668.0A
Other languages
German (de)
French (fr)
Inventor
Agnès CAMUS
Eric Schmitt
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
IMV Technologies SA
Original Assignee
IMV Technologies SA
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by IMV Technologies SA filed Critical IMV Technologies SA
Publication of EP4267302A1 publication Critical patent/EP4267302A1/en
Pending legal-status Critical Current

Links

Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/502715—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/502707—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by the manufacture of the container or its components
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L9/00—Supporting devices; Holding devices
    • B01L9/52—Supports specially adapted for flat sample carriers, e.g. for plates, slides, chips
    • B01L9/527—Supports specially adapted for flat sample carriers, e.g. for plates, slides, chips for microfluidic devices, e.g. used for lab-on-a-chip
    • G01N15/1433—
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Electro-optical investigation, e.g. flow cytometers
    • G01N15/1434—Electro-optical investigation, e.g. flow cytometers using an analyser being characterised by its optical arrangement
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/01—Arrangements or apparatus for facilitating the optical investigation
    • G01N21/03—Cuvette constructions
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/34—Microscope slides, e.g. mounting specimens on microscope slides
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/02—Adapting objects or devices to another
    • B01L2200/025—Align devices or objects to ensure defined positions relative to each other
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/06—Fluid handling related problems
    • B01L2200/0684—Venting, avoiding backpressure, avoid gas bubbles
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/06—Fluid handling related problems
    • B01L2200/0689—Sealing
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/12—Specific details about manufacturing devices
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/04—Closures and closing means
    • B01L2300/041—Connecting closures to device or container
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/06—Auxiliary integrated devices, integrated components
    • B01L2300/0609—Holders integrated in container to position an object
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/06—Auxiliary integrated devices, integrated components
    • B01L2300/0627—Sensor or part of a sensor is integrated
    • B01L2300/0654—Lenses; Optical fibres
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0809—Geometry, shape and general structure rectangular shaped
    • B01L2300/0816—Cards, e.g. flat sample carriers usually with flow in two horizontal directions
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0809—Geometry, shape and general structure rectangular shaped
    • B01L2300/0822—Slides
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
    • B01L2300/088—Channel loops
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0887—Laminated structure
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
    • B01L2400/0406—Moving fluids with specific forces or mechanical means specific forces capillary forces
    • G01N15/01—
    • G01N15/075—
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N2015/1006—Investigating individual particles for cytology
    • G01N2015/1027—
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Electro-optical investigation, e.g. flow cytometers
    • G01N15/1434—Electro-optical investigation, e.g. flow cytometers using an analyser being characterised by its optical arrangement
    • G01N2015/1454—Electro-optical investigation, e.g. flow cytometers using an analyser being characterised by its optical arrangement using phase shift or interference, e.g. for improving contrast
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Electro-optical investigation, e.g. flow cytometers
    • G01N2015/1486—Counting the particles

Definitions

  • TITLE Support for microscopic analysis of a liquid-based biological substance and system comprising such a support and a microscope
  • the invention relates to the microscopic analysis of a liquid-based biological substance, such as animal semen.
  • the invention relates more particularly to a support for such a substance, intended to be mounted in a microscope configured to carry out such an analysis, as well as a system comprising such a support and such a microscope.
  • a drop is deposited at each of the wells of the support.
  • the drops are then each driven by capillary action into the individual compartment with which the well communicates.
  • the holder is mounted in a microscope and the individual compartments are arranged one after another under its lens to observe the substance.
  • a system comprising such a support and such a microscope comprising an objective, 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.
  • the microscope further comprises a frame on which the objective is mounted and on which the stage is mounted movable in translation in the left-right direction of the support in order to be able to successively align the other individual compartments with the objective.
  • the microscope further comprises a support plate and a clamping member configured to urge the support plate and the stage towards each other.
  • the support plate is hinged to the plate so that it can be moved away from the support or folded down on the support.
  • the system is further configured to focus the lens by autofocus.
  • the invention proposes, in a first aspect, a support similar to that described above but with improved performance, in particular vis-à-vis the positioning of the individual compartments with respect to the objective of the microscope.
  • the invention proposes for this purpose a support for the microscopic analysis of a liquid-based biological substance, such as animal semen, comprising a glass slide, a cover glass and lines of glue arranged between the microscope slide and the microscope slide to secure them to one another while peripherally delimiting individual compartments for receiving a sample of said biological substance;
  • the microscope slide being parallelepipedal and having two main faces, namely an upper face and a lower face, and side faces which extend from one to the other of the main 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 being itself greater than the distance between the bottom face and the upper face;
  • said support having a left-right direction transverse to the left face and to the right face while it is parallel to the front face, to the rear face and to the main faces, a front-to-rear direction transverse to the front face and to the rear face while it is parallel to the left face,
  • the expression "uncovered” referring to the upper face of the microscope slide means that this upper face is not in no element (such as the cover glass for example) 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.
  • the invention is further based on the observation that the optical quality of the surface of the microscope 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.
  • 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 placed in abutment by its marginal regions in order to have its mechanically determined position.
  • at least one support element - such as for example a finger as explained later - against which the upper face would be placed in abutment by its marginal regions in order to have its mechanically determined position.
  • the position of the marginal regions - namely along the front face and the rear face - means that these regions are as far apart as possible from each other, which is favorable to the precision of the mechanical determination of the position by the support elements and the stability during sliding.
  • the extension in the left-right direction is equal to the distance between the left face and the right face of the microscope slide;
  • each well has an extension in the front-rear direction which is less than the extension in the front-rear direction of each of said front marginal region and rear marginal region;
  • the distance between the left side and the right side of the microscope slide is between 74.8 mm and 75.2 mm
  • the distance between the front side and the back side of the microscope slide is between 24 .8 mm and 25.2 mm
  • the distance between the left face and the right face of the coverslip is between 31.8 mm and 32.2 mm
  • the distance between the front face and the back face of the coverslip is between 16.8 mm and 16.2 mm;
  • 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 viewing position in which the underside of the microscope slide 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 plate is mounted movable in translation in the left-right direction of the support in order to be able to successively align the other individual compartments with the lens, a support plate extending at least partially opposite the support, and a clamp configured to urge the backing plate and the platen toward each other so that the carrier is clamped between the plate support and the plate; the support plate being fixedly mounted on the frame and comprising a base and support fingers projecting from the base towards the plate and the distal ends of which are in contact with the marginal regions exclusively, with at least two support fingers on one of the front marginal region and rear marginal region and at least one support finger
  • the fingers of support determine the position of the upper face relative to the frame in the bottom-top direction.
  • the position of the upper face relative to the frame does not depend on the thickness (i.e. the distance between the lower face and the upper face) of the slide.
  • the support fingers slide on the marginal regions. Since the extension in the left-right direction of the marginal regions is at least equal to the difference between the respective axes of the two individual compartments furthest apart from each other, the fingers remain in contact with the marginal regions whatever or the individual compartment aligned with the lens.
  • the position of the upper face of the microscope slide in the bottom-up direction remains determined in the same way by the support fingers regardless of the individual compartment aligned with the objective.
  • the mechanical reference surface is the surface of the plate on which the lower face of the slide is resting, so that the position of the upper face of the slide bears -Object depends on the thickness of the slide, and therefore these systems do not offer the advantages described above.
  • the support plate has four support fingers, including two support fingers in contact with the front marginal region and the other two support fingers in contact with the rear marginal region;
  • the clamping device comprises magnets fixed to the other of the plate and the support plate
  • the base has an opening through which the objective points on said individual compartment aligned with the objective in said first observation position, at least two of the support fingers which are in contact with the same region marginal being arranged on either side of the opening;
  • the frame comprises a mounting wall and the base comprises two branches each projecting transversely from the mounting wall, extending opposite one another and delimiting the opening, each branch being provided with two fingers support, including a finger in contact with the front marginal region and a finger in contact with the rear marginal region;
  • the clamping member comprises magnets housed in said branches;
  • one of the plate and of the support plate has a ramp and the other of the plate and of the support plate has a boss configured to cooperate with the ramp during the translational movement of the following plate the left-right direction so as to separate the plate from the support plate, against the force exerted by the clamping member, to a position in which the support is accessible in order to be able to be removed from the platinum;
  • the plate is mechanically connected to the frame by a sliding pivot connection comprising a cylindrical guide longitudinally oriented in the left-right direction and a barrel mounted to slide on the guide, with a clearance between the barrel and the guide; and or
  • the lens has a fixed focal length.
  • 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 comprises a step of counting spermatozoa.
  • FIG. 1 is a view of a support for the analysis by microscopy of a liquid-based biological substance, in accordance with the prior art.
  • FIG. 2 is a view similar to Figure 1, and to the same scale, of a support according to the invention.
  • FIG. 3 is a very schematic sectional view of a system according to the invention (illustrated in more detail in Figures 9 to 12), comprising the support illustrated in Figure 2 and a microscope, the system being in a observation configuration in which the support is clamped between a stage and the fingers of a support plate of the microscope with one of the individual compartments of the support aligned with the objective; the cut being in the front-rear direction of the support and passing through the middle of this individual compartment.
  • FIG. 4 is a sectional view of the system illustrated in Figure 3, this time following the left-right direction of the support and passing through the middle of the individual compartments.
  • Figure 5 is a view similar to Figure 4 after the stage has been moved in the left-right direction to align another of the individual compartments with the lens.
  • Figure 6 is a view similar to Figure 2 on which are marked the axes of the individual compartments, as well as the positions of the fingers of the plate support on the upper face of the specimen slide when the stage is in the position it occupies in figure 4 and when the stage is in the position it occupies in figure 5.
  • Figure 7 is a view similar to Figure 6, on which are marked the path of each of the fingers of the support plate during the passage of the plate from the position it occupies in Figure 4 to the position qu 'it occupies in Figure 5.
  • Figure 8 is a view similar to Figure 3, after the plate has been moved away from the support plate to place the system in a loading/unloading configuration in which the support is accessible to a user of the system.
  • FIG. 9 is an exploded and detailed perspective view of the system illustrated schematically in Figures 2 to 8, but without the lens.
  • FIG. 10 is a view similar to figure 9 but for the system in the assembled state, and in a cleaning configuration in which the plate is very far from the support plate.
  • FIG 11 is a sectional view of the system as illustrated in Figures 9 and 10, taken along the front-rear direction of the support and passing through one of the individual compartments, the system being in the loading / unloading configuration schematically shown in Figure 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 shown in Figures 3 to 5.
  • Figure 13 is a view similar to Figure 7 for a variant of the support according to the invention.
  • the support 10 and the support 40 illustrated respectively in FIGS. 1 and 2 each comprise a microscope slide 11, a microscope slide 12 and lines of glue 13 placed between the microscope slide 11 and the microscope slide. 12 to secure them to each other while peripherally delimiting individual compartments 14. It should be noted beforehand that in this report, the same numerical references have been used for simplicity for the corresponding elements of the support 10 and the support 40.
  • 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.
  • the support 10 here comprises individual compartments 14.
  • 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 lines of glue 13.
  • a drop (not shown) of this substance is deposited in the respective well 15. This drop is then drawn by capillarity into the individual compartment 14 while the air initially present in the individual compartment 14 escapes through the vent 16.
  • the microscope slide 11 and the microscope slide 12 are here each made of float glass coated with a surfactant film (in English surfactant) promoting the entrainment of the drop in the individual compartment 14 by capillarity.
  • a surfactant film in English surfactant
  • the microscope slide 11 and the microscope slide 12 are each transparent.
  • the slide 11 is parallelepipedal and has two main faces, namely an upper face 17 and a lower face 18 (FIGS. 3 and 4), and side faces which extend from one to the other of the faces main ones, namely a left face 19, a right face 20, a front face 21 and a rear face 22.
  • 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 even greater than the distance between the lower face 18 and the upper face 19.
  • the distance between the left face 19 and the right face 20 is here between 74.8 mm and 75.2 mm.
  • the distance between the front face 21 and the rear face 22 is here between 24.8 mm and 25.2 mm.
  • the distance between the lower face 18 and the upper face 19 - that is to say the thickness of the microscope slide 11 - is here approximately 1 mm.
  • the support 10 has a left-right direction transverse to the left face 19 and to the right face 20 while it is parallel to the front face 21, to the rear face 22 and to the main faces 17 and 18, a front- rear transverse to the front face 21 and to the rear face 22 while it is parallel to the left face 19, to the right face 20 and to the main faces 17 and 18, and a low-high direction transverse to the main faces 17 and 18 while it is parallel to the side faces 19, 20, 21 and 22.
  • the coverslip 12 is parallelepipedal and has two main faces, namely an upper face 23 and a lower face 24, and side faces which extend from one to the other of the main faces, namely a face left 25, a right face 26, a front face 27 and a rear face 28.
  • 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.
  • the distance between the left face 25 and the right face 26 is here between 31.8 mm and 32.2 mm.
  • the distance between the front face 27 and the rear face 28 is here between 20.8 mm and 21.2 mm.
  • the distance between the lower face 24 and the upper face 23 - that is to say the thickness of the coverslip 12 - is here 0.7 mm +/- 0.05 mm.
  • the lower face 24 of the coverslip 12 faces 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 slide coverslip 12 are turned towards and at a distance respectively from the front face 21, the rear face 22, the left face 19 and the right face 20 of the slide 11.
  • the cover slip 12 is here arranged centered with respect to the slide 11 in the left-right direction.
  • Each individual compartment 14 is delimited in the bottom-top direction respectively by the upper face 17 of the slide 11 and by the lower face 24 of the slide cover 12 and peripherally delimited by the lines of glue 13.
  • the thickness of the lines of glue 13 is here 20 ⁇ m.
  • Each individual compartment 14 here has a capacity of 3 pl.
  • the individual compartments 14 follow one another in the left-right direction, here being juxtaposed to one another.
  • each individual compartment 14 opens on one side onto a well 15 and on the other side onto a vent 16, the well 15 here being turned towards the rear face 22 of the slide 11 and the vent 16 here being turned towards the front face 21 of the slide 11 .
  • Each well 15 is delimited here by a portion 29 of the lines of glue 13 shaped like a U and extending beyond the space strictly between the slide 11 and the slide cover 12 up to one end 30 located here flush with the rear face 22 of the specimen slide 11 .
  • Each vent 16 is here delimited by straight portions 31 of the lines of glue 13 extending opposite one another, each overflowing the space strictly comprised between the slide 11 and the cover slip. object 12 and up to a respective end 32 located here flush with the front face 21 of the slide 11 .
  • the support 40 illustrated in FIG. 2 is similar to the support 10, except that the respective dimensions of the coverslip 12 and of the glue lines 13 of the support 40 are smaller in the front-to-back direction and that the upper face 17 of the microscope slide 11 of the support 40 is bare and uncovered on a front marginal region 41 and on a rear marginal region 42.
  • 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.
  • 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 cover glass 12, is therefore also smaller than that of the individual compartments 14 of holder 10.
  • the front marginal region 41 runs along the front face 21 of the microscope slide 11, while the rear marginal region 42 runs along the rear face 22 of the microscope slide 11.
  • 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 slide 11 respectively.
  • the front marginal region 41 here has an extension along the front-rear 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 door blade. object 11 .
  • the upper face 17 is coated by the lines of glue 13 and/or is opposite the lower face 24 of the cover slip 12 is n is therefore not bare and uncovered.
  • the rear marginal region 42 here has an extension along the front-rear direction equal to 2.6 mm and has an extension along the left-hand direction. right equal to the distance between the left face 19 and the right face 20 of the specimen slide 11 .
  • the upper face 17 is coated by the lines of glue 13 and/or is opposite the lower face 24 of the cover slip 12 is n is therefore not bare and uncovered.
  • each of the front marginal region 41 and rear marginal region 42 has an extension in the front-rear direction 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.
  • the extension along the front-rear 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 blade covers- object 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 in particular less than both the extension in the front-rear direction of the marginal region front 41 (2.5 mm) and the extension in the front-rear direction of the rear marginal region 42 (2.6 mm).
  • FIGS. 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
  • the system is in an observation configuration in which the microscope 50 is in the process of cooperating with the support 40 in order to observe the samples of biological substance contained in the individual compartments 14.
  • the microscope 50 comprises 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 11 against the stage 52 and one of the individual compartments 14 aligned with the objective
  • the support 40 is in a first viewing position in which the compartment 14 aligned with the objective 51 is the one seen furthest to the left in Figure 2.
  • the microscope 50 is configured to arrange 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.
  • the support 40 is in a second observation position in which the compartment 14 aligned with the lens 51 is the one seen furthest to the right in figure 2.
  • the microscope 50 further comprises a frame (not shown in Figures 3 to 5 and 8 but whose mounting wall 60 is visible in Figures 9 to 11) on which the lens 51 is mounted fixed and on which the plate 52 is mounted movable in translation in the left-right direction of the support 40 in order to be able to successively align the other individual compartments 14 with the lens 51 .
  • the plate 52 is further configured to receive the support 40 in a predetermined position.
  • the plate 52 here comprises positioning stops each configured to cooperate with a respective side face of the slide 11 which it faces in order to immobilize the next support 40 the direction (left-right or front-back depending on the case) transverse this respective side face.
  • the positioning stops here comprise two front positioning stops 33 facing the front side face 21, a left positioning stop 34 facing the left side face 19, a right positioning stop 35 facing the right side face 20 and two rear positioning stops 36 facing the rear side face 22, so that the support 40 is here immobilized with respect to the plate 52 at the same time following the front-back direction in both directions and following the left-right direction in both directions.
  • the positioning stops are here formed by pins projecting from an upper face 37 of the plate 52 on which rests the lower face 18 of the slide 11.
  • the plate 52 further comprises a notch 45 which extends opposite each of the individual compartments 14 and configured to allow light from a lighting device (not shown) arranged under the plate 52 to reach each of the compartments. individual 14.
  • the microscope 50 further comprises a support plate 53 which extends at least partially opposite the support 40, as well as a clamping member configured to urge the support plate 53 and the plate 52 towards one another. other, so that the support 40 is clamped between the support plate 53 and the plate 52.
  • the clamping member here comprises magnets 54 integral with the support plate 53, while the plate 52 has magnetic properties, the magnets 54 being configured to urge the plate 52 towards the support plate 53.
  • the support plate 53 is mounted fixed on the frame and comprises a base 55 and support fingers 56 each projecting from the base 55 towards the plate 52 up to a distal end 57.
  • 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 slide 11.
  • the support plate 53 here comprises four fingers 56, including two fingers 56 which are in contact with the front marginal region 41 and two other fingers 56 which are in contact with the rear marginal region 42.
  • the four support fingers 56 are here arranged so as to be each at the respective vertex of an imaginary rectangle.
  • the base 55 also has an opening 58 through which the lens 51 points to the individual compartment 14 aligned with the lens 51, the latter here being partially received in the opening 58.
  • 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. and other side of opening 58.
  • Lens 51 here has a fixed focal length.
  • System 100 is configured so that focal point 59 of lens 51 is here approximately in the middle of individual compartment 14 which is aligned with lens 51 .
  • the support fingers 56 slide on the respective marginal regions 41 and 42, each remaining exclusively in contact with them.
  • the plate 52 has been moved a distance equal to the difference between the respective axes 38 of the individual compartment 14 the farthest to the left and the individual compartment 14 farthest to the right, that is to say the two individual compartments 14 furthest apart from each other on the support 40.
  • 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 apart from each other is therefore 23.4 mm.
  • FIG. 6 four black rectangles 46 have also identified the respective positions of the support fingers 56 when the plate 52 is in the position it occupies in FIG. 4, and by four hatched rectangles 47 the respective positions of these fingers 56 when the plate 52 is in the position it occupies in Figure 5.
  • Each black 46 or hatched 47 rectangle corresponds to the portion of the upper face 17 which is in contact with the distal end 57 of a bearing finger 56, this portion here having an extension in the front-rear direction equal to approximately 1 mm and an extension along the left-right direction equal to approximately 3 mm.
  • hatched rectangles 48 also identify the path of each of the support fingers 56 when the plate 52 moves from the position it occupies in FIG. 4 to the position it occupies on the figure 5.
  • 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 therefore has here an extension along the front-rear direction equal to approximately 1 mm and an extension along the left-right direction equal to approximately 3 mm added to the length of the path of the finger 56, this latter length being - as explained higher - equal to the difference 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 farthest from each other. the other on support 40.
  • each support finger 56 in order for each support finger 56 to be able to remain entirely in contact with the respective marginal region 41 or 42 which it traverses, whatever the individual compartment 14 which is aligned with the lens 51, the extension following 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.
  • extension in the left-right direction of the hatched rectangles 48 is in practice greater than the difference between the respective axes 38 of the two individual compartments 14 furthest apart from each other on the support 40 (since it is necessary to take into account the extension of the support fingers in the left-right direction).
  • the support has two front marginal regions and/or two rear marginal regions as described above but each with a lesser extension in the left-right direction, and with a coating interposed between these two regions. front marginal regions and/or between these two rear marginal regions.
  • the support 70 shown in Figure 13 is similar to the support 40 except that it has only two individual compartments 14, the gap between the axes 38 of these compartments being in this case equal to 15.84 mm.
  • FIG. 8 illustrates a loading/unloading configuration of the system 100, here obtained after the plate 52 has been moved away from the support plate 53, the support 40 then being accessible to a user to be removed from the plate 52 and possibly replaced by another similar support in order to analyze the samples contained in the compartments of this other support.
  • the plate 52 loaded with the support 40 is brought closer to the support plate 53 until each support finger 56 is at the contact of the marginal regions 41 and 42 in the manner previously described.
  • 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 one another, delimiting the opening 58.
  • Each branch 61 is here provided with two support fingers 56, one finger 56 of which 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 (figure 12).
  • the magnets 54 of the clamping member are here housed in the branches 61, each branch 61 being here provided with three magnets 54 cylindrical.
  • the magnets 54 are here distributed between the bearing fingers 56 of this branch 61 .
  • the support plate 53 here has guide grooves 62 formed 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 33, left 34 and/or right 35 positioning stops 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.
  • the guide grooves 62 thus participate in the correct positioning of the plate 52 in the front-rear 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 movement of plate 52.
  • 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 in the left-right direction of the support 40 so as to separate the plate 52 of 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.
  • the plate 52 is here mechanically connected to the frame by a sliding pivot connection 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.
  • the guide 65 is here fixed to the mounting wall 60 while the barrel 66 is integral with the plate 52.
  • the internal diameter of the barrel 66 is here slightly greater than the external diameter of the guide 65 so that there is a clearance between them allowing the plate 52 to pivot to a certain extent both in the left-right direction in both directions. and following the front-rear direction in both directions.
  • the microscope 50 further comprises here a motorized mechanism (not shown) controlled from a user interface.
  • the mechanism is not motorized and is operated manually using a wheel.
  • the system 100 as described above is in particular suitable for an analysis comprising a step of counting the spermatozoa present in the animal semen samples, with a view to determining the concentration thereof.
  • the system 100 is coupled to a computer configured to automatically analyze the images captured by the lens in order to carry out the counting step and determine the concentration.
  • the focal point of the lens is not in the middle of the individual compartments but is - in the bottom-up direction - at a predetermined distance from the middle of the individual compartments.
  • the lens is mounted to move relative to the frame of the microscope between a plurality of fixed predetermined positions in each of which the lens is at a respective different distance from the support.
  • the term objective must be understood in a broad sense and may in particular be a simple sensor of electronic images, such as for example a CMOS sensor, used for example in so-called “lensless” microscopy methods, using for example the principle of online holography.
  • 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 succession of images in order to analyze the biological substance, for example to determine the motility of spermatozoa contained in animal semen samples.
  • the front marginal strip and the rear marginal strip are located at a short distance from the front face 21 or the rear face 22 respectively, that is to say for example that a coating or a coating portion 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.
  • the support plate comprises three support fingers, with - when the system is in the holding configuration - two of the fingers which are exclusively in contact with one of the front and rear marginal regions and the last of the fingers which is exclusively in contact with each other of the front and back marginal regions.
  • the clamping member comprises magnets which are integral with the plate while the support plate has magnetic properties.
  • the positioning stops are different from pins projecting from 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 blade object holder 11 .
  • the wells and the 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 else compartment to another the wells and the vents are alternately turned towards the front face then towards the rear face.
  • all the wells are turned towards the front face and all the vents are turned towards the rear face, or else compartment to another the wells and the vents are alternately turned towards the front face then towards the rear face.
  • the lens has a variable focal length rather than a fixed one
  • the number of individual compartments is different from four and from two, for example three, five or more than five;
  • the clamping member comprises a number of magnets different from three per branch, for example one, two, four or more than four magnets per branch and/or the magnets have a shape other than cylindrical, for example parallelepipedic;
  • the clamping member is different from magnets housed in the branches of the support plate, and comprises for example springs arranged on the side of the plate which faces away from the support plate and configured to push the plate towards the support plate; and or
  • the ramp 63 is carried by the support plate and the boss 64 is carried by the plate.
  • the boss 64 is carried by the plate.

Abstract

The support comprises an object-carrying slide (11) and a cover glass (12) secured by lines of adhesive (13) peripherally defining individual compartments (14) for receiving a sample of the biological substance; characterised in that the upper face (17) of the object-carrying slide is bare and uncovered over at least one front marginal region (41) and at least one rear marginal region (42), each having an extension in the front-rear direction equal to at least 2.5 mm and an extension in the left-right direction equal to at least the distance between the respective axes of the two individual compartments furthest apart from one another. The system comprises the support (40) and a microscope for enclosing the support between a stage and a support plate provided with fingers, the distal ends of which are in contact with the marginal regions (41, 42) exclusively.

Description

DESCRIPTION DESCRIPTION
TITRE : Support pour l’analyse par microscope d’une substance biologique à base liquide et système comportant un tel support et un microscopeTITLE: Support for microscopic analysis of a liquid-based biological substance and system comprising such a support and a microscope
Domaine technique de l’invention Technical field of the invention
L’invention a trait à l’analyse par microscope d’une substance biologique à base liquide, telle que de la semence animale. L’invention vise plus particulièrement un support pour une telle substance, destiné à être monté dans un microscope configuré pour réaliser une telle analyse, ainsi qu’un système comportant un tel support et un tel microscope. The invention relates to the microscopic analysis of a liquid-based biological substance, such as animal semen. The invention relates more particularly to a support for such a substance, intended to be mounted in a microscope configured to carry out such an analysis, as well as a system comprising such a support and such a microscope.
Etat de la technique State of the art
On connaît déjà, notamment par le brevet européen EP 0 809 815, auquel correspond le brevet américain US 6,551 ,554 ainsi que par le design patent américain US D566,849 S un support pour l’analyse par microscope d’une substance biologique à base liquide, telle que de la semence animale, comportant une lame porte-objet, une lamelle couvre-objet et des lignes de colle disposées entre la lame porte-objet et la lamelle couvre-objet pour les assujettir l’une à l’autre tout en délimitant périphériquement des compartiments individuels de réception d’un échantillon de ladite substance biologique ; la lame porte-objet étant parallélépipédique et présentant deux faces principales, à savoir une face supérieure et une face inférieure, et des faces latérales qui s’étendent de l’une à l’autre des faces principales, à savoir une face gauche, une face droite, une face avant et une face arrière, avec la distance entre la face gauche et la face droite qui est supérieure à la distance entre la face avant et la face arrière qui est elle-même supérieure à la distance entre la face inférieure et la face supérieure ; ledit support présentant une direction gauche- droite transversale à la face gauche et à la face droite tandis qu’elle est parallèle à la face avant, à la face arrière et aux faces principales, une direction avant-arrière transversale à la face avant et à la face arrière tandis qu’elle est parallèle à la face gauche, à la face droite et aux faces principales, et une direction bas-haut transversale aux faces principales tandis qu’elle est parallèle aux faces latérales ; la lamelle couvre-objet étant parallélépipédique et présentant deux faces principales, à savoir une face supérieure et une face inférieure, et des faces latérales qui s’étendent de l’une à l’autre des faces principales, à savoir une face gauche, une face droite, une face avant et une face arrière ; la face inférieure de la lamelle couvre-objet étant en regard de la face supérieure de la lame porte-objet tandis que la face avant, la face arrière, la face gauche et la face droite de la lamelle couvre-objet sont tournées vers et à distance de respectivement la face avant, la face arrière, la face gauche et la face droite de la lame porte-objet ; chaque dit compartiment individuel étant délimité suivant la direction bas-haut respectivement par la face supérieure de la lame porte-objet et par la face inférieure de la lamelle couvre-objet et délimité périphériquement par lesdites lignes de colle, chaque dit compartiment individuel s’ouvrant exclusivement d’un côté sur un puits et d’un autre côté sur un évent, chaque puits et chaque évent étant délimité par lesdites lignes de colle, chaque puits étant tourné vers l’une des face avant et face arrière de la lame porte-objet et chaque évent étant tourné vers l’autre des face avant et face arrière de la lame porte-objet. We already know, in particular by the European patent EP 0 809 815, to which corresponds the American patent US 6,551,554 as well as by the American patent design US D566,849 S a support for the analysis by microscope of a biological substance based liquid, such as animal semen, comprising a microscope slide, a microscope slide and lines of glue disposed between the microscope slide and the microscope slide to secure them together while by peripherally delimiting individual compartments for receiving a sample of said biological substance; the microscope slide being parallelepipedal and having two main faces, namely an upper face and a lower face, and side faces which extend from one to the other of the main 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 being itself greater than the distance between the bottom face and the upper face; said support having a left-right direction transverse to the left face and to the right face while it is parallel to the front face, to the rear face and to the main faces, a front-to-rear direction transverse to the front face and to the rear face while it is parallel to the left face, the right face and the main faces, and a bottom-up direction transverse to the main faces while it is parallel to the side faces; the coverslip being parallelepipedic and having two main faces, namely an upper face and a lower face, and side faces which extend from one to the other of the main faces, namely a left face, a right face, a front face and a rear side; the underside of the coverslip facing the upper side of the coverslip while the front side, the back side, the left side and the right side of the coverslip face towards and away from distance respectively from the front face, the rear face, the left face and the right face of the microscope slide; each said individual compartment being delimited in the bottom-up direction respectively by the upper face of the microscope slide and by the lower face of the microscope slide and peripherally delimited by said lines of glue, each said individual compartment opening exclusively on one side on a well and on the other side on a vent, each well and each vent being delimited by said lines of glue, each well being turned towards one of the front face and rear face of the carrier blade object and each vent being turned towards the other of the front face and rear face of the slide.
Pour analyser la substance biologique, on en dépose une goutte au niveau de chacun des puits du support. Les gouttes sont alors chacune entraînée par capillarité dans le compartiment individuel avec lequel communique le puits. Le support est monté dans un microscope et les compartiments individuels sont disposés l’un après l’autre sous son objectif pour observer la substance. To analyze the biological substance, a drop is deposited at each of the wells of the support. The drops are then each driven by capillary action into the individual compartment with which the well communicates. The holder is mounted in a microscope and the individual compartments are arranged one after another under its lens to observe the substance.
On connaît également un système comportant un tel support et un tel microscope comportant un objectif, une platine sur laquelle repose le support avec la face inférieure de la lame porte-objet contre la platine et un des compartiments individuels aligné avec l’objectif. Le microscope comporte en outre un bâti sur lequel l’objectif est monté et sur lequel la platine est montée mobile en translation suivant la direction gauche-droite du support afin de pouvoir aligner successivement les autres compartiments individuels avec l’objectif. Le microscope comporte en outre une plaque d’appui et un organe de serrage configuré pour solliciter la plaque d’appui et la platine l’une vers l’autre. La plaque d’appui est articulée à la platine pour pouvoir être écartée du support ou rabattue sur le support. Le système est en outre configuré pour effectuer la mise au point de l’objectif par autofocus. A system is also known comprising such a support and such a microscope comprising an objective, 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. The microscope further comprises a frame on which the objective is mounted and on which the stage is mounted movable in translation in the left-right direction of the support in order to be able to successively align the other individual compartments with the objective. The microscope further comprises a support plate and a clamping member configured to urge the support plate and the stage towards each other. The support plate is hinged to the plate so that it can be moved away from the support or folded down on the support. The system is further configured to focus the lens by autofocus.
Exposé de l’invention Disclosure of Invention
L’invention propose, sous un premier aspect, un support similaire à celui décrit ci-dessus mais aux performances améliorées, notamment vis-à-vis du positionnement des compartiments individuels par rapport à l’objectif du microscope. The invention proposes, in a first aspect, a support similar to that described above but with improved performance, in particular vis-à-vis the positioning of the individual compartments with respect to the objective of the microscope.
L’invention propose à cet effet un support pour l’analyse par microscope d’une substance biologique à base liquide, telle que de la semence animale, comportant une lame porte-objet, une lamelle couvre-objet et des lignes de colle disposées entre la lame porte-objet et la lamelle couvre-objet pour les assujettir l’une à l’autre tout en délimitant périphériquement des compartiments individuels de réception d’un échantillon de ladite substance biologique ; la lame porte-objet étant parallélépipédique et présentant deux faces principales, à savoir une face supérieure et une face inférieure, et des faces latérales qui s’étendent de l’une à l’autre des faces principales, à savoir une face gauche, une face droite, une face avant et une face arrière, avec la distance entre la face gauche et la face droite qui est supérieure à la distance entre la face avant et la face arrière qui est elle-même supérieure à la distance entre la face inférieure et la face supérieure ; ledit support présentant une direction gauche-droite transversale à la face gauche et à la face droite tandis qu’elle est parallèle à la face avant, à la face arrière et aux faces principales, une direction avant-arrière transversale à la face avant et à la face arrière tandis qu’elle est parallèle à la face gauche, à la face droite et aux faces principales, et une direction bas-haut transversale aux faces principales tandis qu’elle est parallèle aux faces latérales ; la lamelle couvre-objet étant parallélépipédique et présentant deux faces principales, à savoir une face supérieure et une face inférieure, et des faces latérales qui s’étendent de l’une à l’autre des faces principales, à savoir une face gauche, une face droite, une face avant et une face arrière ; la face inférieure de la lamelle couvre-objet étant en regard de la face supérieure de la lame porte-objet tandis que la face avant, la face arrière, la face gauche et la face droite de la lamelle couvre-objet sont tournées vers et à distance de respectivement la face avant, la face arrière, la face gauche et la face droite de la lame porte-objet ; chaque dit compartiment individuel étant délimité suivant la direction bas-haut respectivement par la face supérieure de la lame porte-objet et par la face inférieure de la lamelle couvre- objet et délimité périphériquement par lesdites lignes de colle, chaque dit compartiment individuel s’ouvrant exclusivement d’un côté sur un puits et d’un autre côté sur un évent, chaque puits et chaque évent étant délimité par lesdites lignes de colle, chaque puits étant tourné vers l’une des face avant et face arrière de la lame porte-objet et chaque évent étant tourné vers l’autre des face avant et face arrière de la lame porte-objet ; caractérisé en ce que la face supérieure de la lame porte-objet est à nu et à découvert sur au-moins une région marginale avant et sur au-moins une région marginale arrière, la région marginale avant longeant la face avant de la lame porte-objet, la région marginale arrière longeant la face arrière de la lame porte-objet, chacune desdites région marginale avant et région marginale arrière ayant une extension suivant la direction avant-arrière au-moins égale à 2,5 mm et une extension suivant la direction gauche-droite au moins égale à l’écart entre les axes respectifs des deux compartiments individuels les plus éloignés l’un de l’autre et au plus égale à la distance entre la face gauche et la face droite de la lame porte-objet. The invention proposes for this purpose a support for the microscopic analysis of a liquid-based biological substance, such as animal semen, comprising a glass slide, a cover glass and lines of glue arranged between the microscope slide and the microscope slide to secure them to one another while peripherally delimiting individual compartments for receiving a sample of said biological substance; the microscope slide being parallelepipedal and having two main faces, namely an upper face and a lower face, and side faces which extend from one to the other of the main 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 being itself greater than the distance between the bottom face and the upper face; said support having a left-right direction transverse to the left face and to the right face while it is parallel to the front face, to the rear face and to the main faces, a front-to-rear direction transverse to the front face and to the rear face while it is parallel to the left face, the right face and the main faces, and a bottom-up direction transverse to the main faces while it is parallel to the side faces; the coverslip being parallelepipedal and having two main faces, namely an upper face and a lower face, and side faces which extend from one to the other of the main faces, namely a left face, a right side, a front side and a back side; the lower face of the coverslip facing the upper face of the coverslip while the front face, the rear face, the left face and the right face of the coverslip are facing towards and away from respectively the front face, the rear face, the left face and the right face of the slide; each said individual compartment being delimited in the bottom-up direction respectively by the upper face of the object slide and by the underside of the cover glass and peripherally delimited by said lines of glue, each said individual compartment opening exclusively on one side on a well and on the other side on a vent, each well and each vent being delimited by said lines of glue, each well being turned towards one of the front face and rear face of the carrier blade object and each vent being turned towards the other of the front face and rear face of the slide; characterized in that the upper face of the slide is bare and uncovered over at least one front marginal region and over at least one rear marginal region, the front marginal region running along the front face of the slide object, 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 in the front-rear direction at least equal to 2.5 mm and an extension in the direction left-right at least equal to the difference between the respective axes of the two individual compartments farthest from each other and at most equal to the distance between the left face and the right face of the slide.
Grâce aux régions marginales, il est possible d’effectuer un positionnement du support selon l’invention vis-à-vis d’un microscope par appui sur la face supérieure de la lame porte-objet, et non sur sa face inférieure comme cela se fait classiquement. Thanks to the marginal regions, it is possible to carry out a positioning of the support according to the invention vis-à-vis a microscope by pressing on the upper face of the slide, and not on its lower face as it is classically done.
Cela permet un positionnement particulièrement précis suivant la direction bas-haut de l’échantillon à analyser, puisque l’on évite ainsi l’incertitude sur l’épaisseur de la lame porte-objet due aux tolérances de fabrication. This allows particularly precise positioning along the bottom-top direction of the sample to be analyzed, since uncertainty about the thickness of the microscope slide due to manufacturing tolerances is thus avoided.
On notera que l’expression « à nu » visant la face supérieure de la lame porte-objet signifie que cette face supérieure ne porte directement sur elle aucun revêtement ajouté localement, tel que les lignes de colle ou de l’encre. It should be noted that the expression "naked" referring to the upper face of the microscope slide means that this upper face does not bear directly on it any coating added locally, such as lines of glue or ink.
On notera également que l’expression « à découvert » visant la face supérieure de la lame porte-objet signifie que cette face supérieure n’est en regard d’aucun élément (comme par exemple la lame couvre-objet) qui pourrait constituer un obstacle pour un autre élément qui serait rapproché du support suivant la direction bas-haut pour arriver au contact de sa face supérieure. It will also be noted that the expression "uncovered" referring to the upper face of the microscope slide means that this upper face is not in no element (such as the cover glass for example) 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.
Grâce aux régions marginales, il est possible d’accéder directement à la face supérieure de la lame porte-objet afin d’en déterminer mécaniquement la position et ainsi déterminer la position d’autres éléments dont la position est fixe et/ou connue par rapport à cette face supérieure. On notera en particulier que déterminer mécaniquement la position de la face supérieure revient à déterminer mécaniquement la position des compartiments individuels que la face supérieure délimite. Thanks to the marginal regions, it is possible to directly access the upper face of the microscope slide in order to mechanically determine its position and thus determine the position of other elements whose position is fixed and/or known with respect to to this upper side. It will be noted in particular that mechanically determining the position of the upper face amounts to mechanically determining the position of the individual compartments that the upper face delimits.
L’invention est en outre basée sur l’observation que la qualité optique de la surface de la lame porte-objet (nécessaire afin de perturber le moins possible la lumière qui traverse la lame pour illuminer l’échantillon présent dans le compartiment individuel) implique une régularité mécanique de cette surface qui est favorable au glissement ; et qu’il est notamment possible d’exercer un appui sur cette surface tout en conservant une capacité de glissement sur cette surface. The invention is further based on the observation that the optical quality of the surface of the microscope 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.
Les régions marginales avant et arrière peuvent ainsi servir de pistes de glissement pour au moins un élément d’appui - tel que par exemple un doigt comme expliqué plus loin - contre lequel la face supérieure serait mise en appui par ses régions marginales afin d’avoir sa position déterminée mécaniquement. 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 placed in abutment by its marginal regions in order to have its mechanically determined position.
On notera en outre que la position des régions marginales - à savoir le long de la face avant et de la face arrière - fait que ces régions sont le plus à l’écart possible l’une de l’autre, ce qui est favorable à la précision de la détermination mécanique du positionnement par les éléments d’appui et à la stabilité lors du glissement. It will 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 as possible from each other, which is favorable to the precision of the mechanical determination of the position by the support elements and the stability during sliding.
S’agissant de la borne inférieure de l’extension des régions marginales suivant la direction avant-arrière - à savoir 2,5 mm -, la demanderesse a déterminé que, compte tenu des tolérances de fabrication des différents éléments du système, une valeur au-moins égale à 2,5 mm permettait de garantir que les doigts d’appui restent exclusivement au contact des régions marginales avant et arrière, quelle que soit la position de la platine suivant la direction gauche-droite. With regard to the lower limit of the extension of the marginal regions in the front-rear direction - namely 2.5 mm -, the applicant has determined that, taking into account the manufacturing tolerances of the various elements of the system, a value at -less equal to 2.5 mm made it possible to guarantee that the support fingers remained exclusively in contact with the regions front and rear marginals, whatever the position of the plate in the left-right direction.
Des caractéristiques avantageuses particulièrement simples, commodes et économiques du support selon l’invention sont présentées ci- dessous : Particularly simple, convenient and economical advantageous characteristics of the support according to the invention are presented below:
- pour l’une au moins de la région marginale avant et de la région marginale arrière, l’extension suivant la direction gauche-droite est égale à la distance entre la face gauche et la face droite de la lame porte-objet ; - for at least one of the front marginal region and the rear marginal region, the extension in the left-right direction is equal to the distance between the left face and the right face of the microscope slide;
- chaque puits a une extension suivant la direction avant-arrière qui est inférieure à l’extension suivant la direction avant-arrière de chacune desdites région marginale avant et région marginale arrière ; - each well has an extension in the front-rear direction which is less than the extension in the front-rear direction of each of said front marginal region and rear marginal region;
- la distance entre la face gauche et la face droite de la lame porte-objet est comprise entre 74,8 mm et 75,2 mm, la distance entre la face avant et la face arrière de la lame porte-objet est comprise entre 24,8 mm et 25,2 mm, la distance entre la face gauche et la face droite de la lamelle couvre-objet est comprise entre 31 ,8 mm et 32,2 mm, et la distance entre la face avant et la face arrière de la lamelle couvre-objet est comprise entre 16,8 mm et 16,2 mm; et/ou- the distance between the left side and the right side of the microscope slide is between 74.8 mm and 75.2 mm, the distance between the front side and the back side of the microscope slide is between 24 .8 mm and 25.2 mm, the distance between the left face and the right face of the coverslip is between 31.8 mm and 32.2 mm, and the distance between the front face and the back face of the coverslip is between 16.8 mm and 16.2 mm; and or
- tous les puits sont tournés vers l’une des face avant et face arrière de la lame porte-objet et tous les évents sont tournés vers l’autre des face avant et face arrière de la lame porte-objet. - all the wells are turned towards one of the front faces and rear faces of the microscope slide and all the vents are turned towards the other front face and rear face of the microscope slide.
L’invention propose, sous un deuxième aspect, un système pour l’analyse d’une substance biologique à base liquide, comportant un support tel que décrit ci- dessus et un microscope comportant un objectif, une platine configurée pour recevoir le support dans une première position d’observation dans laquelle la face inférieure de la lame porte-objet est contre la platine et un des compartiments individuels est aligné avec l’objectif, un bâti sur lequel l’objectif est monté dans au moins une position prédéterminée fixe et sur lequel la platine est montée mobile en translation suivant la direction gauche-droite du support afin de pouvoir aligner successivement les autres compartiments individuels avec l’objectif, une plaque d’appui s’étendant au moins partiellement en regard du support, et un organe de serrage configuré pour solliciter la plaque d’appui et la platine l’une vers l’autre de sorte que le support est enserré entre la plaque d’appui et la platine ; la plaque d’appui étant montée fixe sur le bâti et comportant une embase et des doigts d’appui saillant de l’embase vers la platine et dont les extrémités distales sont au contact des régions marginales exclusivement, avec au moins deux doigts d’appui sur l’une des région marginale avant et région marginale arrière et au moins un doigt d’appui sur l’autre des région marginale avant et région marginale arrière ; la platine présentant des butées de positionnement pour immobiliser le support par rapport à la platine suivant la direction gauche-droite. 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 viewing position in which the underside of the microscope slide 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 plate is mounted movable in translation in the left-right direction of the support in order to be able to successively align the other individual compartments with the lens, a support plate extending at least partially opposite the support, and a clamp configured to urge the backing plate and the platen toward each other so that the carrier is clamped between the plate support and the plate; the support plate being fixedly mounted on the frame and comprising a base and support fingers projecting from the base towards the plate and the distal ends of which are in contact with the marginal regions exclusively, 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.
Puisque la plaque d’appui est montée fixe sur le bâti et que la face supérieure de la lame porte-objet est maintenue contre les doigts d’appui par la platine, sous l’effet de l’organe de serrage, les doigts d’appui déterminent la position de la face supérieure par rapport au bâti suivant la direction bas-haut. Since the support plate is mounted fixed on the frame and the upper face of the slide is held against the support fingers by the plate, under the effect of the clamping member, the fingers of support determine the position of the upper face relative to the frame in the bottom-top direction.
On notera qu’ainsi la position de la face supérieure par rapport au bâti ne dépend pas de l’épaisseur (c’est-à-dire la distance entre la face inférieure et la face supérieure) de la lame porte-objet. It should be noted that thus the position of the upper face relative to the frame does not depend on the thickness (i.e. the distance between the lower face and the upper face) of the slide.
Lors de la translation de la platine suivant la direction gauche-droite, les doigts d’appui glissent sur les régions marginales. Puisque l’extension suivant la direction gauche-droite des régions marginales est au moins égale à l’écart entre les axes respectifs des deux compartiments individuels les plus éloignés l’un de l’autre, les doigts restent au contact des régions marginales quel que soit le compartiment individuel aligné avec l’objectif. During the translation of the stage in the left-right direction, the support fingers slide on the marginal regions. Since the extension in the left-right direction of the marginal regions is at least equal to the difference between the respective axes of the two individual compartments furthest apart from each other, the fingers remain in contact with the marginal regions whatever or the individual compartment aligned with the lens.
Ainsi, la position de la face supérieure de la lame porte-objet suivant la direction bas-haut reste déterminée de la même manière par les doigts d’appui quel que soit le compartiment individuel aligné avec l’objectif. Thus, the position of the upper face of the microscope slide in the bottom-up direction remains determined in the same way by the support fingers regardless of the individual compartment aligned with the objective.
Ceci permet notamment, une fois que la mise au point de l’objectif a été faite pour un compartiment individuel donné, de conserver cette mise au point pour chacun des autres compartiments individuels. This makes it possible in particular, once the focus of the objective has been made for a given individual compartment, to retain this focus for each of the other individual compartments.
Ceci permet en outre de conserver cette mise au point pour un autre support mis en remplacement d’un premier support pour lequel la mise au point a été effectuée, puisque la position suivant la direction bas-haut de la face supérieure de cet autre support sera déterminée par les doigts d’appui de la même manière que pour le premier support. On notera qu’un telle mise au point peut alors correspondre à un réglage d’usine effectué lors de la fabrication du microscope. This also makes it possible to keep this focusing for another support placed in replacement of a first support for which the focusing has been carried out, since the position according to 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. It will be noted that such a focusing can then correspond to a factory adjustment carried out during the manufacture of the microscope.
On notera encore que dans les systèmes de l’art antérieur, la surface de référence mécanique est la surface de la platine sur laquelle repose la face inférieure de la lame porte-objet, de sorte que la position de la face supérieure de la lame porte-objet dépend de l’épaisseur de la lame porte-objet, et que par conséquent ces systèmes n’offrent pas les avantages exposés ci-dessus. It will also be noted that in the systems of the prior art, the mechanical reference surface is the surface of the plate on which the lower face of the slide is resting, so that the position of the upper face of the slide bears -Object depends on the thickness of the slide, and therefore these systems do not offer the advantages described above.
Des caractéristiques avantageuses particulièrement simples, commodes et économiques du système selon l’invention sont présentées ci- dessous : Particularly simple, convenient and economical advantageous characteristics of the system according to the invention are presented below:
- la plaque d’appui comporte quatre doigts d’appui, dont deux doigts d’appui au contact de la région marginale avant et les deux autres doigts d’appui au contact de la région marginale arrière ; - the support plate has four support fingers, including two support fingers in contact with the front marginal region and the other two support fingers in contact with the rear marginal region;
- l’un de la platine et de la plaque d’appui a des propriétés magnétiques et l’organe de serrage comporte des aimants solidaires de l’autre de la platine et de la plaque d’appui ; - one of the plate and the support plate has magnetic properties and the clamping device comprises magnets fixed to the other of the plate and the support plate;
- l’embase présente une ouverture au travers de laquelle l’objectif pointe sur ledit compartiment individuel aligné avec l’objectif dans ladite première position d’observation, au-moins deux des doigts d’appui qui sont au contact d’une même région marginale étant disposés de part et d’autre de l’ouverture ; - the base has an opening through which the objective points on said individual compartment aligned with the objective in said first observation position, at least two of the support fingers which are in contact with the same region marginal being arranged on either side of the opening;
- le bâti comporte une paroi de montage et l’embase comporte deux branches saillant chacune transversalement de la paroi de montage, s’étendant en regard l’une de l’autre et délimitant l’ouverture, chaque branche étant pourvue de deux doigts d’appui, dont un doigt au contact de la région marginale avant et un doigt au contact de la région marginale arrière ; - the frame comprises a mounting wall and the base comprises two branches each projecting transversely from the mounting wall, extending opposite one another and delimiting the opening, each branch being provided with two fingers support, including a finger in contact with the front marginal region and a finger in contact with the rear marginal region;
- l’organe de serrage comporte des aimants logés dans lesdites branches ;- the clamping member comprises magnets housed in said branches;
- l’une de la platine et de la plaque d’appui présente une rampe et l’autre de la platine et de la plaque d’appui présente un bossage configuré pour coopérer avec la rampe au cours du mouvement de translation de la platine suivant la direction gauche-droite de manière à écarter la platine de la plaque d’appui, à l’encontre de la force exercée par l’organe de serrage, jusqu’à une position dans laquelle le support est accessible pour pouvoir être retiré de la platine ; - la platine est mécaniquement reliée au bâti par une liaison pivot glissant comportant un guide cylindrique longitudinalement orienté suivant la direction gauche-droite et un fût monté à coulissement sur le guide, avec un jeu entre le fût et le guide ; et/ou - one of the plate and of the support plate has a ramp and the other of the plate and of the support plate has a boss configured to cooperate with the ramp during the translational movement of the following plate the left-right direction so as to separate the plate from the support plate, against the force exerted by the clamping member, to a position in which the support is accessible in order to be able to be removed from the platinum; - The plate is mechanically connected to the frame by a sliding pivot connection comprising a cylindrical guide longitudinally oriented in the left-right direction and a barrel mounted to slide on the guide, with a clearance between the barrel and the guide; and or
- l’objectif a une distance focale fixe. - the lens has a fixed focal length.
L’invention vise également l’utilisation d’un système tel qu’exposé ci- dessus dans laquelle la substance biologique à base liquide est de la semence animale et ladite analyse comprend une étape de comptage des spermatozoïdes. 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 comprises a step of counting spermatozoa.
Brève description des figures Brief description of figures
On va maintenant poursuivre l'exposé de l'invention par la description détaillée d'exemples de réalisation, donnée ci-après à titre illustratif et non limitatif, en référence aux dessins annexés. Sur ceux-ci : We will now continue the description of the invention with the detailed description of exemplary embodiments, given below by way of non-limiting illustration, with reference to the appended drawings. On these:
- La figure 1 est une vue d’un support pour l’analyse par microscopie d’une substance biologique à base liquide, conforme à l’art antérieur. - Figure 1 is a view of a support for the analysis by microscopy of a liquid-based biological substance, in accordance with the prior art.
- La figure 2 est une vue similaire à la figure 1 , et à la même échelle, d’un support conforme à l’invention. - Figure 2 is a view similar to Figure 1, and to the same scale, of a support according to the invention.
- La figure 3 est une vue en coupe très schématique d’un système conforme à l’invention (illustré plus en détails sur les figures 9 à 12), comportant le support illustré sur la figure 2 et un microscope, le système étant dans une configuration d’observation dans laquelle le support est enserré entre une platine et les doigts d’une plaque d’appui du microscope avec un des compartiment individuels du support aligné avec l’objectif ; la coupe étant suivant la direction avant-arrière du support et passant par le milieu de ce compartiment individuel. - Figure 3 is a very schematic sectional view of a system according to the invention (illustrated in more detail in Figures 9 to 12), comprising the support illustrated in Figure 2 and a microscope, the system being in a observation configuration in which the support is clamped between a stage and the fingers of a support plate of the microscope with one of the individual compartments of the support aligned with the objective; the cut being in the front-rear direction of the support and passing through the middle of this individual compartment.
- La figure 4 est une vue en coupe du système illustré à la figure 3, cette fois suivant la direction gauche-droite du support et passant par le milieu des compartiments individuels. - Figure 4 is a sectional view of the system illustrated in Figure 3, this time following the left-right direction of the support and passing through the middle of the individual compartments.
- La figure 5 est une vue similaire à la figure 4 après que la platine a été déplacée suivant la direction gauche-droite pour aligner un autre des compartiments individuels avec l’objectif. - Figure 5 is a view similar to Figure 4 after the stage has been moved in the left-right direction to align another of the individual compartments with the lens.
- La figure 6 est une vue similaire à la figure 2 sur laquelle sont repérés les axes des compartiments individuels, ainsi que les positions des doigts de la plaque d’appui sur la face supérieure de la lame porte-objet lorsque la platine est dans la position qu’elle occupe à la figure 4 et lorsque la platine est dans la position qu’elle occupe à la figure 5. - Figure 6 is a view similar to Figure 2 on which are marked the axes of the individual compartments, as well as the positions of the fingers of the plate support on the upper face of the specimen slide when the stage is in the position it occupies in figure 4 and when the stage is in the position it occupies in figure 5.
- La figure 7 est une vue similaire à la figure 6, sur laquelle sont repérées le trajet de chacun des doigts de la plaque d’appui lors du passage de la platine de la position qu’elle occupe sur la figure 4 à la position qu’elle occupe sur la figure 5. - Figure 7 is a view similar to Figure 6, on which are marked the path of each of the fingers of the support plate during the passage of the plate from the position it occupies in Figure 4 to the position qu 'it occupies in Figure 5.
- La figure 8 est une vue similaire à la figure 3, après que la platine a été écartée de la plaque d’appui pour passer le système dans une configuration de chargement/déchargement dans laquelle le support est accessible à un utilisateur du système. - Figure 8 is a view similar to Figure 3, after the plate has been moved away from the support plate to place the system in a loading/unloading configuration in which the support is accessible to a user of the system.
- La figure 9 est une vue en perspective éclatée et détaillée du système illustré schématiquement sur les figures 2 à 8, mais sans l’objectif. - Figure 9 is an exploded and detailed perspective view of the system illustrated schematically in Figures 2 to 8, but without the lens.
- La figure 10 est une vue semblable à la figure 9 mais pour le système à l’état assemblé, et dans une configuration de nettoyage dans laquelle la platine est très à l’écart de la plaque d’appui. - Figure 10 is a view similar to figure 9 but for the system in the assembled state, and in a cleaning configuration in which the plate is very far from the support plate.
- La figure 11 est une vue en coupe du système tel qu’illustré sur les figures 9 et 10, prise suivant la direction avant-arrière du support et passant par un des compartiments individuels, le système étant dans la configuration de chargement/déchargement schématiquement représentée à la figure 8. - Figure 11 is a sectional view of the system as illustrated in Figures 9 and 10, taken along the front-rear direction of the support and passing through one of the individual compartments, the system being in the loading / unloading configuration schematically shown in Figure 8.
- La figure 12 est une vue de côté d’une partie du système illustré sur les figures 9 à 11 , le système étant dans la configuration d’observation schématiquement représentée sur les figures 3 à 5. - Figure 12 is a side view of part of the system illustrated in Figures 9 to 11, the system being in the observation configuration schematically shown in Figures 3 to 5.
- La figure 13 est une vue similaire à la figure 7 pour une variante du support conforme à l’invention. - Figure 13 is a view similar to Figure 7 for a variant of the support according to the invention.
Description détaillée detailed description
Le support 10 et le support 40 illustrés respectivement sur les figures 1 et 2 comportent chacun une lame porte-objet 11 , une lamelle couvre-objet 12 et des lignes de colle 13 disposées entre la lame porte-objet 11 et la lamelle couvre-objet 12 pour les assujettir l’une à l’autre tout en délimitant périphériquement des compartiments individuels 14. On notera au préalable que dans le présent rapport, on a employé pour simplifier les mêmes références numériques pour les éléments correspondants du support 10 et du support 40. The support 10 and the support 40 illustrated respectively in FIGS. 1 and 2 each comprise a microscope slide 11, a microscope slide 12 and lines of glue 13 placed between the microscope slide 11 and the microscope slide. 12 to secure them to each other while peripherally delimiting individual compartments 14. It should be noted beforehand that in this report, the same numerical references have been used for simplicity for the corresponding elements of the support 10 and the support 40.
Par conséquent, dans la description du support 10 qui suit, s’il est fait référence à un élément du support 10 qui n’est pas visible sur la figure 1 , il est possible - à des fins de compréhension - de se référer à l’une des figures illustrant le support 40 sur laquelle l’élément correspondant est visible. Consequently, in the description of the support 10 which follows, if reference is made to an element of the support 10 which is not visible in FIG. one of the figures illustrating the support 40 on which the corresponding element is visible.
Les compartiments individuels 14 du support 10 sont chacun configurés pour recevoir un échantillon de la substance biologique à base liquide, ici de la semence animale, diluée ou non. 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.
Le support 10 comporte ici compartiments individuels 14.The support 10 here comprises individual compartments 14.
Chaque compartiment individuel 14 s’ouvre exclusivement d’un côté sur un puits 15 et d’un autre côté sur un évent 16, chaque puits 15 et chaque évent 16 étant délimité par les lignes de colle 13. 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 lines of glue 13.
Pour remplir un compartiment individuel 14 avec un échantillon de la substance biologique, une goutte (non illustrée) de cette substance est déposée dans le puits 15 respectif. Cette goutte est alors entraînée par capillarité dans le compartiment individuel 14 tandis que l’air initialement présent dans le compartiment individuel 14 s’échappe par l’évent 16. 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 capillarity into the individual compartment 14 while the air initially present in the individual compartment 14 escapes through the vent 16.
La lame porte-objet 11 et la lamelle couvre-objet 12 sont ici chacune en verre flotté revêtu d’un film tensioactif (en anglais surfactant) favorisant l’entrainement de la goutte dans le compartiment individuel 14 par capillarité. The microscope slide 11 and the microscope slide 12 are here each made of float glass coated with a surfactant film (in English surfactant) promoting the entrainment of the drop in the individual compartment 14 by capillarity.
La lame porte-objet 11 et la lamelle couvre-objet 12 sont chacune transparente. The microscope slide 11 and the microscope slide 12 are each transparent.
La lame porte-objet 11 est parallélépipédique et présente deux faces principales, à savoir une face supérieure 17 et une face inférieure 18 (figures 3 et 4), et des faces latérales qui s’étendent de l’une à l’autre des faces principales, à savoir une face gauche 19, une face droite 20, une face avant 21 et une face arrière 22. The slide 11 is parallelepipedal and has two main faces, namely an upper face 17 and a lower face 18 (FIGS. 3 and 4), and side faces which extend from one to the other of the faces main ones, namely a left face 19, a right face 20, a front face 21 and a rear face 22.
La distance entre la face gauche 19 et la face droite 20 est supérieure à la distance entre la face avant 21 et la face arrière 22 qui est elle- même supérieure à la distance entre la face inférieure 18 et la face supérieure 19. 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 even greater than the distance between the lower face 18 and the upper face 19.
La distance entre la face gauche 19 et la face droite 20 est ici comprise entre 74,8 mm et 75,2 mm. The distance between the left face 19 and the right face 20 is here between 74.8 mm and 75.2 mm.
La distance entre la face avant 21 et la face arrière 22 est ici comprise entre 24,8 mm et 25,2 mm. The distance between the front face 21 and the rear face 22 is here between 24.8 mm and 25.2 mm.
La distance entre la face inférieure 18 et la face supérieure 19 - c’est-à-dire l’épaisseur de la lame porte-objet 11 - est ici d’environ 1 mm. The distance between the lower face 18 and the upper face 19 - that is to say the thickness of the microscope slide 11 - is here approximately 1 mm.
Le support 10 présente une direction gauche-droite transversale à la face gauche 19 et à la face droite 20 tandis qu’elle est parallèle à la face avant 21 , à la face arrière 22 et aux faces principales 17 et 18, une direction avant- arrière transversale à la face avant 21 et à la face arrière 22 tandis qu’elle est parallèle à la face gauche 19, à la face droite 20 et aux faces principales 17 et 18, et une direction bas-haut transversale aux faces principales 17 et 18 tandis qu’elle est parallèle aux faces latérales 19, 20, 21 et 22. The support 10 has a left-right direction transverse to the left face 19 and to the right face 20 while it is parallel to the front face 21, to the rear face 22 and to the main faces 17 and 18, a front- rear transverse to the front face 21 and to the rear face 22 while it is parallel to the left face 19, to the right face 20 and to the main faces 17 and 18, and a low-high direction transverse to the main faces 17 and 18 while it is parallel to the side faces 19, 20, 21 and 22.
La lamelle couvre-objet 12 est parallélépipédique et présente deux faces principales, à savoir une face supérieure 23 et une face inférieure 24, et des faces latérales qui s’étendent de l’une à l’autre des faces principales, à savoir une face gauche 25, une face droite 26, une face avant 27 et une face arrière 28. The coverslip 12 is parallelepipedal and has two main faces, namely an upper face 23 and a lower face 24, and side faces which extend from one to the other of the main faces, namely a face left 25, a right face 26, a front face 27 and a rear face 28.
La distance entre la face gauche 25 et la face droite 26 est supérieure à la distance entre la face avant 27 et la face arrière 28 qui est elle- même supérieure à la distance entre la face inférieure 24 et la face supérieure 23. 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.
La distance entre la face gauche 25 et la face droite 26 est ici comprise entre 31 ,8 mm et 32,2 mm. The distance between the left face 25 and the right face 26 is here between 31.8 mm and 32.2 mm.
La distance entre la face avant 27 et la face arrière 28 est ici comprise entre 20,8 mm et 21 ,2 mm. The distance between the front face 27 and the rear face 28 is here between 20.8 mm and 21.2 mm.
La distance entre la face inférieure 24 et la face supérieure 23 - c’est-à-dire l’épaisseur de la lamelle couvre-objet 12 - est ici de 0,7 mm +/- 0,05 mm. La face inférieure 24 de la lamelle couvre-objet 12 est en regard de la face supérieure 17 de la lame porte-objet 11 tandis que la face avant 27, la face arrière 28, la face gauche 25 et la face droite 26 de la lamelle couvre-objet 12 sont tournées vers et à distance de respectivement la face avant 21 , la face arrière 22, la face gauche 19 et la face droite 20 de la lame porte-objet 11 . The distance between the lower face 24 and the upper face 23 - that is to say the thickness of the coverslip 12 - is here 0.7 mm +/- 0.05 mm. The lower face 24 of the coverslip 12 faces 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 slide coverslip 12 are turned towards and at a distance respectively from the front face 21, the rear face 22, the left face 19 and the right face 20 of the slide 11.
La lamelle couvre-objet 12 est ici disposée centrée par rapport à la lame porte-objet 11 suivant la direction gauche-droite. The cover slip 12 is here arranged centered with respect to the slide 11 in the left-right direction.
Chaque compartiment individuel 14 est délimité suivant la direction bas-haut respectivement par la face supérieure 17 de la lame porte-objet 11 et par la face inférieure 24 de la lamelle couvre-objet 12 et délimité périphériquement par les lignes de colle 13. Each individual compartment 14 is delimited in the bottom-top direction respectively by the upper face 17 of the slide 11 and by the lower face 24 of the slide cover 12 and peripherally delimited by the lines of glue 13.
L’épaisseur des lignes de colle 13 est ici de 20 pm. The thickness of the lines of glue 13 is here 20 μm.
On notera que sur les figures 3 à 5 et 8, l’épaisseur des lignes de colle 13 a été exagérée par souci de lisibilité. It will be noted that in FIGS. 3 to 5 and 8, the thickness of the lines of glue 13 has been exaggerated for the sake of readability.
Chaque compartiment individuel 14 a ici une contenance de 3 pl.Each individual compartment 14 here has a capacity of 3 pl.
Les compartiments individuels 14 se succèdent suivant la direction gauche-droite, ici en étant juxtaposés l’un à l’autre. The individual compartments 14 follow one another in the left-right direction, here being juxtaposed to one another.
Comme expliqué plus haut, chaque compartiment individuel 14 s’ouvre d’un côté sur un puits 15 et d’un autre côté sur un évent 16, le puits 15 étant ici tourné vers la face arrière 22 de la lame porte-objet 11 et l’évent 16 étant ici tourné vers la face avant 21 de la lame porte-objet 11 . 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 here being turned towards the rear face 22 of the slide 11 and the vent 16 here being turned towards the front face 21 of the slide 11 .
Ici tous les puits 15 sont tournés vers la face arrière 22 et tous les évents 16 sont tournés vers la face avant 21 . Here all the wells 15 face the rear face 22 and all the vents 16 face the front face 21 .
Chaque puits 15 est ici délimité par une portion 29 des lignes de colle 13 conformée en U et s’étendant en débord de l’espace strictement compris entre la lame porte-objet 11 et la lamelle couvre-objet 12 jusqu’à une extrémité 30 située ici au raz de la face arrière 22 de la lame porte-objet 11 . Each well 15 is delimited here by a portion 29 of the lines of glue 13 shaped like a U and extending beyond the space strictly between the slide 11 and the slide cover 12 up to one end 30 located here flush with the rear face 22 of the specimen slide 11 .
Chaque évent 16 est ici délimité par des portions droites 31 des lignes de colle 13 s’étendant en regard l’une de l’autre, chacune en débord de l’espace strictement compris entre la lame porte-objet 11 et la lamelle couvre- objet 12 et jusqu’à une extrémité 32 respective située ici au raz de la face avant 21 de la lame porte-objet 11 . Le support 40 illustré sur la figure 2 est similaire au support 10, sauf que les dimensions respectives de la lamelle couvre-objet 12 et des lignes de colle 13 du support 40 sont plus petites suivant la direction avant-arrière et que la face supérieure 17 de la lame porte-objet 11 du support 40 est à nu et à découvert sur une région marginale avant 41 et sur une région marginale arrière 42. Each vent 16 is here delimited by straight portions 31 of the lines of glue 13 extending opposite one another, each overflowing the space strictly comprised between the slide 11 and the cover slip. object 12 and up to a respective end 32 located here flush with the front face 21 of the slide 11 . The support 40 illustrated in FIG. 2 is similar to the support 10, except that the respective dimensions of the coverslip 12 and of the glue lines 13 of the support 40 are smaller in the front-to-back direction and that the upper face 17 of the microscope slide 11 of the support 40 is bare and uncovered on a front marginal region 41 and on a rear marginal region 42.
Plus précisément, la distance entre la face avant 27 et la face arrière 28 de la lamelle couvre-objet 12 est ici comprise entre 16,8 mm et 16,2 mm. More precisely, 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.
On notera que la longueur des compartiments individuels 14 du support 40, c’est-à-dire la distance séparant le puits 15 de l’évent 16, qui correspond ici à la distance entre la face avant 27 et la face arrière 28 de la lamelle couvre-objet 12, est par conséquent elle aussi plus petite que celle des compartiments individuels 14 du support 10. 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 cover glass 12, is therefore also smaller than that of the individual compartments 14 of holder 10.
La région marginale avant 41 longe la face avant 21 de la lame porte-objet 11 , tandis que la région marginale arrière 42 longe la face arrière 22 de la lame porte-objet 11 . The front marginal region 41 runs along the front face 21 of the microscope slide 11, while the rear marginal region 42 runs along the rear face 22 of the microscope slide 11.
Ici, la région marginale avant 41 et la région marginale arrière 42 sont chacune directement bordée par la face avant 21 et la face arrière 22 de la lame porte-objet 11 respectivement. 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 slide 11 respectively.
La région marginale avant 41 a ici une extension suivant la direction avant-arrière égale à 2,5 mm et a une extension suivant la direction gauche- droite égale à la distance entre la face gauche 19 et la face droite 20 de la lame porte-objet 11 . The front marginal region 41 here has an extension along the front-rear 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 door blade. object 11 .
La limite de l’extension suivant la direction avant-arrière de la région marginale avant 41 est repérée sur la figure 2 par le trait interrompu 43, où elle passe au raz de chacune des extrémités 32 des évents 16. The limit of the extension along the front-rear direction of the front marginal region 41 is marked in FIG. 2 by the broken line 43, where it passes flush with each of the ends 32 of the vents 16.
En effet, au-delà de cette limite 43 en allant vers l’arrière, la face supérieure 17 est revêtue par les lignes de colle 13 et/ou se trouve en regard de la face inférieure 24 de la lamelle couvre-objet 12 est n’est donc pas à nu et à découvert. Indeed, beyond this limit 43 going towards the rear, the upper face 17 is coated by the lines of glue 13 and/or is opposite the lower face 24 of the cover slip 12 is n is therefore not bare and uncovered.
La région marginale arrière 42 a ici une extension suivant la direction avant-arrière égale à 2,6 mm et a une extension suivant la direction gauche- droite égale à la distance entre la face gauche 19 et la face droite 20 de la lame porte-objet 11 . The rear marginal region 42 here has an extension along the front-rear direction equal to 2.6 mm and has an extension along the left-hand direction. right equal to the distance between the left face 19 and the right face 20 of the specimen slide 11 .
La limite de l’extension suivant la direction avant-arrière de la région marginale arrière 42 est repérée sur la figure 2 par le trait interrompu 44, où elle passe au raz des extrémités 30 des puits 15. The limit of the extension along the front-rear direction of the rear marginal region 42 is marked in FIG. 2 by the broken line 44, where it passes flush with the ends 30 of the wells 15.
En effet, au-delà de cette limite 44 en allant vers l’avant, la face supérieure 17 est revêtue par les lignes de colle 13 et/ou se trouve en regard de la face inférieure 24 de la lamelle couvre-objet 12 est n’est donc pas à nu et à découvert. Indeed, beyond this limit 44 going forwards, the upper face 17 is coated by the lines of glue 13 and/or is opposite the lower face 24 of the cover slip 12 is n is therefore not bare and uncovered.
D’une manière générale, chacune des région marginale avant 41 et région marginale arrière 42 a une extension suivant la direction avant-arrière au plus égale à 3 mm, ou en variante égale à plus que 3 mm, par exemple 3,5 mm, 4 mm, 4,5 mm, 5 mm, voire plus que 5 mm. In general, each of the front marginal region 41 and rear marginal region 42 has an extension in the front-rear direction 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.
En outre, on notera que l’extension suivant la direction avant-arrière de chacun des puits 15 - c’est-à-dire la distance, mesurée au niveau de la face supérieure 17, entre la face arrière 28 de la lame couvre-objet 12 et la limite 44 (qui passe au raz des extrémités 30 des puits 15) - est ici égale à 2,4 mm, ce qui est notamment inférieur à la fois à l’extension suivant la direction avant- arrière de la région marginale avant 41 (2,5 mm) et à l’extension suivant la direction avant-arrière de la région marginale arrière 42 (2,6 mm). In addition, it will be noted that the extension along the front-rear 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 blade covers- object 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 in particular less than both the extension in the front-rear direction of the marginal region front 41 (2.5 mm) and the extension in the front-rear direction of the rear marginal region 42 (2.6 mm).
Les figures 3 à 5 et 8 illustrent un système 100 pour l’analyse de la substance biologique à base liquide, comportant le support 40 et un microscopeFIGS. 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. 50.
Sur les figures 3 à 5, le système est dans une configuration d’observation dans laquelle le microscope 50 est en cours de coopération avec le support 40 afin d’observer les échantillons de substance biologique contenus dans les compartiments individuels 14. In FIGS. 3 to 5, the system is in an observation configuration in which the microscope 50 is in the process of cooperating with the support 40 in order to observe the samples of biological substance contained in the individual compartments 14.
Le microscope 50 comporte un objectif 51 et une platine 52 configurée pour recevoir le support 40 dans au-moins une position d’observation dans laquelle la face inférieure 18 de la lame porte-objet 11 contre la platine 52 et un des compartiments individuels 14 aligné avec l’objectifThe microscope 50 comprises 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 11 against the stage 52 and one of the individual compartments 14 aligned with the objective
51. Sur les figures 3 et 4, le support 40 est dans une première position d’observation dans laquelle le compartiment 14 aligné avec l’objectif 51 est celui que l’on voit le plus à gauche sur la figure 2. 51. In Figures 3 and 4, the support 40 is in a first viewing position in which the compartment 14 aligned with the objective 51 is the one seen furthest to the left in Figure 2.
Comme expliqué plus en détails ci-après, le microscope 50 est configuré pour disposer le support 40 dans une pluralité de positions d’observation dans chacune desquelles la platine 52 reçoit le support 40 avec un compartiment individuel 14 respectif aligné avec l’objectif 51. As explained in more detail below, the microscope 50 is configured to arrange 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.
Sur la figure 5 par exemple, le support 40 est dans une deuxième position d’observation dans laquelle le compartiment 14 aligné avec l’objectif 51 est celui que l’on voit le plus à droite sur la figure 2. In figure 5 for example, the support 40 is in a second observation position in which the compartment 14 aligned with the lens 51 is the one seen furthest to the right in figure 2.
Le microscope 50 comporte en outre un bâti (non illustré sur les figures 3 à 5 et 8 mais dont une paroi de montage 60 est visible sur les figures 9 à 11 ) sur lequel l’objectif 51 est monté fixe et sur lequel la platine 52 est montée mobile en translation suivant la direction gauche-droite du support 40 afin de pouvoir aligner successivement les autres compartiments individuels 14 avec l’objectif 51 . The microscope 50 further comprises a frame (not shown in Figures 3 to 5 and 8 but whose mounting wall 60 is visible in Figures 9 to 11) on which the lens 51 is mounted fixed and on which the plate 52 is mounted movable in translation in the left-right direction of the support 40 in order to be able to successively align the other individual compartments 14 with the lens 51 .
La liaison mécanique entre la platine 52 et le bâti sera décrite en détail ultérieurement, à l’appui des figures 9 à 12. The mechanical connection between the plate 52 and the frame will be described in detail later, with the support of figures 9 to 12.
La platine 52 est en outre configurée pour recevoir le support 40 dans une position prédéterminée. The plate 52 is further configured to receive the support 40 in a predetermined position.
Pour positionner et immobiliser le support 40 dans cette position prédéterminée, la platine 52 comporte ici des butées de positionnement chacune configurée pour coopérer avec une face latérale respective de la lame porte-objet 11 dont elle est en regard afin d’immobiliser le support 40 suivant la direction (gauche-droite ou avant-arrière suivant le cas) transversale cette face latérale respective. To position and immobilize the support 40 in this predetermined position, the plate 52 here comprises positioning stops each configured to cooperate with a respective side face of the slide 11 which it faces in order to immobilize the next support 40 the direction (left-right or front-back depending on the case) transverse this respective side face.
Les butées de positionnement comportent ici deux butées de positionnement avant 33 en regard de la face latérale avant 21 , une butée de positionnement gauche 34 en regard de la face latérale gauche 19, une butée de positionnement droite 35 en regard de la face latérale droite 20 et deux butées de positionnement arrière 36 en regard de la face latérale arrière 22, de sorte que le support 40 est ici immobilisé par rapport à la platine 52 à la fois suivant la direction avant-arrière dans les deux sens et suivant la direction gauche-droite dans les deux sens. The positioning stops here comprise two front positioning stops 33 facing the front side face 21, a left positioning stop 34 facing the left side face 19, a right positioning stop 35 facing the right side face 20 and two rear positioning stops 36 facing the rear side face 22, so that the support 40 is here immobilized with respect to the plate 52 at the same time following the front-back direction in both directions and following the left-right direction in both directions.
Les butées de positionnement sont ici formées par des pions saillant d’une face supérieure 37 de la platine 52 sur laquelle repose la face inférieure 18 de la lame porte-objet 11 . The positioning stops are here formed by pins projecting from an upper face 37 of the plate 52 on which rests the lower face 18 of the slide 11.
La platine 52 comporte en outre une échancrure 45 qui s’étend en regard de chacun des compartiments individuels 14 et configurée pour permettre à la lumière d’un organe d’éclairage (non illustré) disposé sous la platine 52 d’atteindre chacun des compartiments individuels 14. The plate 52 further comprises a notch 45 which extends opposite each of the individual compartments 14 and configured to allow light from a lighting device (not shown) arranged under the plate 52 to reach each of the compartments. individual 14.
Le microscope 50 comporte en outre une plaque d’appui 53 qui s’étend au moins partiellement en regard du support 40, ainsi qu’un organe de serrage configuré pour solliciter la plaque d’appui 53 et la platine 52 l’une vers l’autre, de sorte que le support 40 est enserré entre la plaque d’appui 53 et la platine 52. The microscope 50 further comprises a support plate 53 which extends at least partially opposite the support 40, as well as a clamping member configured to urge the support plate 53 and the plate 52 towards one another. other, so that the support 40 is clamped between the support plate 53 and the plate 52.
L’organe de serrage comporte ici des aimants 54 solidaires de la plaque d’appui 53, tandis que la platine 52 a des propriétés magnétiques, les aimants 54 étant configurés pour solliciter la platine 52 vers la plaque d’appui 53. The clamping member here comprises magnets 54 integral with the support plate 53, while the plate 52 has magnetic properties, the magnets 54 being configured to urge the plate 52 towards the support plate 53.
La plaque d’appui 53 est montée fixe sur le bâti et comporte une embase 55 et des doigts d’appui 56 saillant chacun de l’embase 55 vers la platine 52 jusqu’à une extrémité distale 57. The support plate 53 is mounted fixed on the frame and comprises a base 55 and support fingers 56 each projecting from the base 55 towards the plate 52 up to a distal end 57.
Les extrémités distales 57 des doigts d’appui 56 sont chacune exclusivement au contact des régions marginales 41 et 42 de la face supérieure 17 de la lame porte-objet 11 . 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 slide 11.
La lame porte-objet 11 est ainsi enserrée entre les doigts d’appui 56 et la platine 52, la lame porte-objet 11 étant au contact de la plaque d’appui 53 ici exclusivement par l’intermédiaire des doigts d’appui 56. Slide 11 is thus clamped between support fingers 56 and plate 52, slide 11 being in contact with support plate 53 here exclusively via support fingers 56.
La plaque d’appui 53 comporte ici quatre doigts 56, dont deux doigts 56 qui sont au contact de la région marginale avant 41 et deux autres doigts 56 qui sont au contact de la région marginale arrière 42. The support plate 53 here comprises four fingers 56, including two fingers 56 which are in contact with the front marginal region 41 and two other fingers 56 which are in contact with the rear marginal region 42.
Les quatre doigts d’appui 56 sont ici disposés de sorte à se trouver chacun au sommet respectif d’un rectangle imaginaire. L’embase 55 présente en outre une ouverture 58 au travers de laquelle l’objectif 51 pointe sur le compartiment individuel 14 aligné avec l’objectif 51 , ce dernier étant ici partiellement reçu dans l’ouverture 58. The four support fingers 56 are here arranged so as to be each at the respective vertex of an imaginary rectangle. The base 55 also has an opening 58 through which the lens 51 points to the individual compartment 14 aligned with the lens 51, the latter here being partially received in the opening 58.
Les doigts d’appui 56 qui sont au contact de la région marginale avant 41 sont disposés de part et d’autre de l’ouverture 58. Les doigts d’appui 56 qui sont au contact de la région marginale arrière 42 sont disposés de part et d’autre de l’ouverture 58. 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. and other side of opening 58.
L’objectif 51 a ici une distance focale fixe. Lens 51 here has a fixed focal length.
Le système 100 est configuré de sorte que le point focal 59 de l’objectif 51 se trouve ici approximativement au milieu du compartiment individuel 14 qui est aligné avec l’objectif 51 . System 100 is configured so that focal point 59 of lens 51 is here approximately in the middle of individual compartment 14 which is aligned with lens 51 .
Lors de la translation de la platine 52 suivant la direction gauche- droite du support 40, les doigts d’appui 56 glissent sur les régions marginales 41 et 42 respectives, en restant chacun exclusivement à leur contact. During the translation of the plate 52 in 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.
Sur la figure 6, on a repéré les axes 38 de chacun des compartiments individuels 14. In Figure 6, the axes 38 of each of the individual compartments 14 have been identified.
Lorsqu’on aligne un compartiment individuel 14 avec l’objectif 51 pour observer l’échantillon reçu dans ce compartiment 14, on aligne plus précisément l’axe 38 de ce compartiment 14 avec l’axe optique de l’objectif 51 . 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 aligned more precisely with the optical axis of the objective 51 .
Par conséquent, entre la position qu’elle occupe sur la figure 4 et la position qu’elle occupe sur la figure 5, la platine 52 a été déplacée sur une distance égale à l’écart entre les axes 38 respectifs du compartiment individuel 14 le plus à gauche et le compartiment individuel 14 le plus à droite, c’est-à-dire les deux compartiments individuels 14 les plus éloignés l’un de l’autre sur le support 40. Therefore, between the position it occupies in Figure 4 and the position it occupies in Figure 5, the plate 52 has been moved a distance equal to the difference between the respective axes 38 of the individual compartment 14 the farthest to the left and the individual compartment 14 farthest to the right, that is to say the two individual compartments 14 furthest apart from each other on the support 40.
L’écart entre les axes 38 de deux compartiments voisins est ici égal à 7,8 mm et l’écart entre les axes 38 des deux compartiments individuels 14 les plus éloignés l’un de l’autre est donc de 23,4 mm. 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 apart from each other is therefore 23.4 mm.
Sur la figure 6, on a en outre repéré par quatre rectangles noirs 46 les positions respectives des doigts d’appui 56 lorsque la platine 52 est dans la position qu’elle occupe sur la figure 4, et par quatre rectangles hachurés 47 les positions respectives de ces doigts 56 lorsque la platine 52 est dans la position qu’elle occupe sur la figure 5. In FIG. 6, four black rectangles 46 have also identified the respective positions of the support fingers 56 when the plate 52 is in the position it occupies in FIG. 4, and by four hatched rectangles 47 the respective positions of these fingers 56 when the plate 52 is in the position it occupies in Figure 5.
Chaque rectangle noir 46 ou hachuré 47 correspond à la portion de la face supérieure 17 qui est en contact avec l’extrémité distale 57 d’un doigt d’appui 56, cette portion ayant ici une extension suivant la direction avant- arrière égale à environ 1 mm et une extension suivant la direction gauche-droite égale à environ 3 mm. Each black 46 or hatched 47 rectangle corresponds to the portion of the upper face 17 which is in contact with the distal end 57 of a bearing finger 56, this portion here having an extension in the front-rear direction equal to approximately 1 mm and an extension along the left-right direction equal to approximately 3 mm.
Sur la figure 7, on a en outre repéré par des rectangles hachurés 48 le trajet de chacun des doigts d’appui 56 lorsque la platine 52 passe de la position qu’elle occupe sur la figure 4 à la position qu’elle occupe sur la figure 5. In FIG. 7, hatched rectangles 48 also identify the path of each of the support fingers 56 when the plate 52 moves from the position it occupies in FIG. 4 to the position it occupies on the figure 5.
Chaque rectangle hachuré 48 correspond donc à l’intégrale du rectangle 46 ou 47 respectif sur l’ensemble du trajet du doigt 56 correspondant. Chaque rectangle 48 a donc ici une extension suivant la direction avant-arrière égale à environ 1 mm et une extension suivant la direction gauche-droite égale à environ 3 mm additionnée de la longueur du trajet du doigt 56, cette dernière longueur étant - comme expliqué plus haut - égale à l’écart entre les axes 38 respectifs du compartiment individuel 14 le plus à gauche et le compartiment individuel 14 le plus à droite, c’est-à-dire les deux compartiments individuels 14 les plus éloignés l’un de l’autre sur le support 40. 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 therefore has here an extension along the front-rear direction equal to approximately 1 mm and an extension along the left-right direction equal to approximately 3 mm added to the length of the path of the finger 56, this latter length being - as explained higher - equal to the difference 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 farthest from each other. the other on support 40.
On notera que pour que chaque doigt d’appui 56 puisse rester entièrement au contact de la région marginale 41 ou 42 respective qu’il parcourt, quel que soit le compartiment individuel 14 qui est aligné avec l’objectif 51 , l’extension suivant la direction gauche-droite de cette région marginale doit être au-moins égale à l’extension suivant la direction gauche- droite des rectangles hachurés 48. It will be noted that in order for each support finger 56 to be able to remain entirely in contact with the respective marginal region 41 or 42 which it traverses, whatever the individual compartment 14 which is aligned with the lens 51, the extension following 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.
On notera encore que l’extension suivant la direction gauche-droite des rectangles hachurés 48 est en pratique supérieure à l’écart entre les axes 38 respectifs des deux compartiments individuels 14 les plus éloignés l’un de l’autre sur le support 40 (puisqu’il faut tenir compte de l’extension des doigts d’appui suivant la direction gauche-droite). It will also be noted that the extension in the left-right direction of the hatched rectangles 48 is in practice greater than the difference between the respective axes 38 of the two individual compartments 14 furthest apart from each other on the support 40 ( since it is necessary to take into account the extension of the support fingers in the left-right direction).
On notera encore que sur la figure 7, les rectangles hachurés 48 d’une même région marginale 41 ou 42 ne se recoupent pas, de sorte qu’un revêtement pourrait être présent entre ces deux rectangles 48 sans être rencontré par un doigt d’appui 56 au cours de son trajet. Ainsi, dans une variante non illustrée, le support présente deux régions marginales avant et/ou deux régions marginales arrière telle que décrites ci-dessus mais chacune avec une extension moindre suivant la direction gauche-droite, et avec un revêtement interposé entre ces deux régions marginales avant et/ou entre ces deux régions marginales arrière. It will 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 a variant not shown, the support has two front marginal regions and/or two rear marginal regions as described above but each with a lesser extension in the left-right direction, and with a coating interposed between these two regions. front marginal regions and/or between these two rear marginal regions.
Le support 70 illustré à la figure 13 est similaire au support 40 sauf qu’il ne comporte que deux compartiments individuels 14, l’écart entre les axes 38 de ces compartiments étant dans ce cas égal à 15,84 mm. The support 70 shown in Figure 13 is similar to the support 40 except that it has only two individual compartments 14, the gap between the axes 38 of these compartments being in this case equal to 15.84 mm.
La figure 8 illustre une configuration de chargement/déchargement du système 100, ici obtenue après que la platine 52 a été écartée de la plaque d’appui 53, le support 40 étant alors accessible à un utilisateur pour être retiré de la platine 52 et éventuellement remplacé par un autre support similaire afin d’analyser les échantillons contenus dans les compartiments de cet autre support. FIG. 8 illustrates a loading/unloading configuration of the system 100, here obtained after the plate 52 has been moved away from the support plate 53, the support 40 then being accessible to a user to be removed from the plate 52 and possibly replaced by another similar support in order to analyze the samples contained in the compartments of this other support.
Inversement, pour passer le système 100 de la configuration de chargement/déchargement à la configuration d’observation, la platine 52 chargée du support 40 est rapprochée de la plaque d’appui 53 jusqu’à ce que chaque doigt d’appui 56 soient au contact des régions marginales 41 et 42 de la manière décrite précédemment. Conversely, to pass the system 100 from the loading/unloading configuration to the observation configuration, the plate 52 loaded with the support 40 is brought closer to the support plate 53 until each support finger 56 is at the contact of the marginal regions 41 and 42 in the manner previously described.
On va maintenant décrire plus en détail le système 100 à l’appui des figures 9 à 12. We will now describe the system 100 in more detail with the support of FIGS. 9 to 12.
Le bâti comporte ici une paroi de montage 60 et l’embase 55 comporte deux branches 61 saillant chacune transversalement de la paroi de montage 60. Les branches 61 s’étendent en regard l’une de l’autre en délimitant l’ouverture 58. 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 one another, delimiting the opening 58.
Chaque branche 61 est ici pourvue de deux doigts d’appui 56, dont un doigt 56 est au contact de la région marginale avant 41 et un doigt est au contact de la région marginale arrière 42 lorsque le système 100 est dans la configuration d’observation (figure 12). Comme on le voit bien sur la figure 10, les aimants 54 de l’organe de serrage sont ici logés dans les branches 61 , chaque branche 61 étant ici pourvue de trois aimants 54 cylindriques. Pour chaque branche 61 , les aimants 54 sont ici répartis entre les doigts d’appui 56 de cette branche 61 . Each branch 61 is here provided with two support fingers 56, one finger 56 of which 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 (figure 12). As clearly seen in Figure 10, the magnets 54 of the clamping member are here housed in the branches 61, each branch 61 being here provided with three magnets 54 cylindrical. For each branch 61, the magnets 54 are here distributed between the bearing fingers 56 of this branch 61 .
La plaque d’appui 53 présente ici des rainures de guidage 62 ménagées dans l’embase 55, du même côté que les doigts d’appui 56, chaque rainure de guidage 62 étant configurée pour recevoir une portion d’extrémité d’un pion respectif formant l’une des butées de positionnement avant 33, gauche 34 et/ou droite 35 de la platine 52 lorsque le système 100 est en configuration d’observation, les extrémités des pions se déplaçant dans la rainure de guidage respective lorsque la platine 52 se déplace suivant la direction gauche-droite. The support plate 53 here has guide grooves 62 formed 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 33, left 34 and/or right 35 positioning stops 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.
Les rainures de guidage 62 participent ainsi au bon positionnement de la platine 52 suivant la direction avant-arrière du support 40, c’est-à-dire afin que les doigts d’appui 56 restent exclusivement au contact des régions marginales 41 et 42 lors du déplacement de la platine 52. The guide grooves 62 thus participate in the correct positioning of the plate 52 in the front-rear 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 movement of plate 52.
La platine 52 présente en outre une rampe 63 et la plaque d’appui 53 présente un bossage 64 configuré pour coopérer avec la rampe 63 au cours du mouvement de translation de la platine 52 suivant la direction gauche-droite du support 40 de manière à écarter la platine 52 de la plaque d’appui 53, à l’encontre de la force exercée par l’organe de serrage, ici jusqu’à ce que la platine arrive dans la position illustrée sur la figure 11 , où le système 100 est dans sa configuration de chargement/déchargement. 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 in the left-right direction of the support 40 so as to separate the plate 52 of 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.
La platine 52 est ici mécaniquement reliée au bâti par une liaison pivot glissant comportant un guide cylindrique 65 et un fût 66 monté à coulissement sur le guide 65, ce dernier étant longitudinalement orienté suivant une direction parallèle à la direction gauche-droite du support 40 lorsque ce dernier est reçu dans sa position prédéterminée sur la platine 52. The plate 52 is here mechanically connected to the frame by a sliding pivot connection 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.
Un telle liaison mécanique permet à la fois d’éloigner et rapprocher la platine 52 de la plaque d’appui 53 et de déplacer la platine 52 suivant la direction gauche-droite. Le guide 65 est ici fixé à la paroi de montage 60 tandis que le fût 66 est solidaire de la platine 52. Such a mechanical connection makes it possible both to move the plate 52 away from and bring it closer to the support plate 53 and to move the plate 52 in the left-right direction. The guide 65 is here fixed to the mounting wall 60 while the barrel 66 is integral with the plate 52.
Le diamètre interne du fût 66 est ici légèrement supérieur au diamètre externe du guide 65 de sorte qu’il existe entre eux un jeu permettant à la platine 52 de pivoter dans une certaine mesure à la fois suivant la direction gauche-droite dans les deux sens et suivant la direction avant-arrière dans les deux sens. The internal diameter of the barrel 66 is here slightly greater than the external diameter of the guide 65 so that there is a clearance between them allowing the plate 52 to pivot to a certain extent both in the left-right direction in both directions. and following the front-rear direction in both directions.
Cette capacité de la platine 52 à pivoter permet d’ajuster l’inclinaison de la face supérieure 17 de la lame porte-objet 11 par rapport aux doigts d’appui 56 afin de s’assurer que ces derniers soient tous au contact de la face supérieure 17, avec un appui équitablement réparti entre tous les doigts 56. This ability of the plate 52 to pivot makes it possible to adjust the inclination of the upper face 17 of the slide 11 with respect to the support fingers 56 in order to ensure that the latter are all in contact with the face. upper 17, with support evenly distributed between all the fingers 56.
Pour déplacer la platine 52 suivant la direction gauche-droite, le microscope 50 comporte en outre ici un mécanisme motorisé (non illustré) commandé depuis une interface utilisateur. En variante, le mécanisme n’est pas motorisé et est actionné manuellement à l’aide d’une molette. To move the stage 52 in the left-right direction, the microscope 50 further comprises here a motorized mechanism (not shown) controlled from a user interface. Alternatively, the mechanism is not motorized and is operated manually using a wheel.
Le système 100 tel que décrit ci-dessus est notamment adapté pour une analyse comportant une étape de comptage des spermatozoïdes présents dans les échantillons de semence animale, en vue d’en déterminer la concentration. The system 100 as described above is in particular suitable for an analysis comprising a step of counting the spermatozoa present in the animal semen samples, with a view to determining the concentration thereof.
Dans une variante non illustrée, le système 100 est couplé à un ordinateur configuré pour analyser automatiquement les images captées par l’objectif afin de réaliser l’étape de comptage et déterminer la concentration. In a variant not shown, the system 100 is coupled to a computer configured to automatically analyze the images captured by the lens in order to carry out the counting step and determine the concentration.
Dans une variante non illustrée, le point focal de l’objectif n’est pas au milieu des compartiments individuels mais se trouve - suivant la direction bas-haut - à une distance prédéterminée du milieu des compartiments individuels. In a variant not shown, the focal point of the lens is not in the middle of the individual compartments but is - in the bottom-up direction - at a predetermined distance from the middle of the individual compartments.
Dans une variante non illustrée, l’objectif est monté mobile par rapport au bâti du microscope entre une pluralité de positions prédéterminées fixes dans chacune desquelles l’objectif se trouve à une distance respective du support différente. In a variant not shown, the lens is mounted to move relative to the frame of the microscope between a plurality of fixed predetermined positions in each of which the lens is at a respective different distance from the support.
Il est entendu que dans la présente demande, le terme objectif doit être compris dans un sens large et peut notamment être un simple capteur d’images électronique, comme par exemple un capteur CMOS, utilisé par exemple dans les procédés de microscopie dit « sans lentille », utilisant par exemple le principe de l'holographie en ligne. Ainsi, en variante, le système d’analyse tel que décrit ci-dessus est pourvu d’une caméra équipée d’un tel capteur électronique, et est configuré pour enregistrer et traiter une succession d’image afin d’analyser la substance biologique, par exemple pour déterminer la motilité des spermatozoïdes contenus dans des échantillons de semence animale. It is understood that in the present application, the term objective must be understood in a broad sense and may in particular be a simple sensor of electronic images, such as for example a CMOS sensor, used for example in so-called “lensless” microscopy methods, using for example the principle of online holography. Thus, as a variant, 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 succession of images in order to analyze the biological substance, for example to determine the motility of spermatozoa contained in animal semen samples.
Dans une autre variante non illustrée, la bande marginale avant et la bande marginale arrière se trouvent à une faible distance de la face avant 21 ou de la face arrière 22 respectivement, c’est-à-dire par exemple qu’un revêtement ou une portion de revêtement se trouve sur la face supérieure 17 entre la face avant 21 et la région marginale avant et/ou entre la face arrière 22 et la région marginale arrière. In another variant not shown, the front marginal strip and the rear marginal strip are located at a short distance from the front face 21 or the rear face 22 respectively, that is to say for example that a coating or a coating portion 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.
Dans encore une autre variante non illustrée, la plaque d’appui comporte trois doigts d’appui, avec - lorsque le système est en configuration de maintien - deux des doigts qui sont exclusivement au contact de l’une des régions marginales avant et arrière et le dernier des doigts qui est exclusivement au contact de l’autre des régions marginales avant et arrière. In yet another variant not shown, the support plate comprises three support fingers, with - when the system is in the holding configuration - two of the fingers which are exclusively in contact with one of the front and rear marginal regions and the last of the fingers which is exclusively in contact with each other of the front and back marginal regions.
Dans encore une autre variante non illustrée, l’organe de serrage comporte des aimants qui sont solidaires de la platine tandis que la plaque d’appui a des propriétés magnétiques. In yet another variant not shown, the clamping member comprises magnets which are integral with the plate while the support plate has magnetic properties.
Dans encore une autre variante non illustrée, les butées de positionnement sont différentes de pions saillant de la face supérieure 37 de la platine 52 et sont par exemple formées par les bords d’un renfoncement ménagé dans cette face supérieure 37 et configuré pour accueillir la lame porte- objet 11 . In yet another variant not shown, the positioning stops are different from pins projecting from 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 blade object holder 11 .
Dans encore d’autres variantes non illustrées, les puits et les évents sont disposés différemment que décrit ci-dessus, par exemple tous les puits sont tournés vers la face avant et tous les évents sont tournés vers la face arrière, ou bien d’un compartiment à l’autre les puits et les évents sont en alternance tournés vers la face avant puis vers la face arrière. Dans d’autres variantes non illustrées : In still other variants not illustrated, the wells and the 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 else compartment to another the wells and the vents are alternately turned towards the front face then towards the rear face. In other variants not shown:
- l’objectif a une distance focale variable plutôt que fixe ; - the lens has a variable focal length rather than a fixed one;
- le nombre de compartiments individuels est différent de quatre et de deux, par exemple trois, cinq ou plus que cinq ; - l’organe de serrage comporte un nombre d’aimants différent de trois par branche, par exemple un, deux, quatre ou plus de quatre aimants par branche et/ou les aimants ont une forme différente de cylindrique, par exemple parallélépipédique ; - the number of individual compartments is different from four and from two, for example three, five or more than five; - the clamping member comprises a number of magnets different from three per branch, for example one, two, four or more than four magnets per branch and/or the magnets have a shape other than cylindrical, for example parallelepipedic;
- l’organe de serrage est différent d’aimants logés dans les branches de la plaque d’appui, et comporte par exemple des ressorts disposés du côté de la platine qui regarde à l’opposé de la plaque d’appui et configurés pour pousser la platine vers la plaque d’appui ; et/ou - the clamping member is different from magnets housed in the branches of the support plate, and comprises for example springs arranged on the side of the plate which faces away from the support plate and configured to push the plate towards the support plate; and or
- la rampe 63 est portée par la plaque d’appui et le bossage 64 est porté par la platine. De nombreuses autres variantes sont possibles en fonction des circonstances et l’on rappelle à cet égard que l’invention ne se limite pas aux exemples décrits et représentés. - the ramp 63 is carried by the support plate and the boss 64 is carried by the plate. Many other variants are possible depending on the circumstances and it is recalled in this respect that the invention is not limited to the examples described and represented.

Claims

REVENDICATIONS
1 . Support pour l’analyse par microscopie d’une substance biologique à base liquide, telle que de la semence animale, comportant une lame porte-objet (11 ), une lamelle couvre-objet (12) et des lignes de colle (13) disposées entre la lame porte-objet (11 ) et la lamelle couvre-objet (12) pour les assujettir l’une à l’autre tout en délimitant périphériquement des compartiments individuels (14) de réception d’un échantillon de ladite substance biologique ; la lame porte-objet (11 ) étant parallélépipédique et présentant deux faces principales, à savoir une face supérieure (17) et une face inférieure (18), et des faces latérales qui s’étendent de l’une à l’autre des faces principales, à savoir une face gauche (19), une face droite (20), une face avant (21 ) et une face arrière (22), avec la distance entre la face gauche (19) et la face droite (20) qui est supérieure à la distance entre la face avant (21 ) et la face arrière (22) qui est elle-même supérieure à la distance entre la face inférieure (18) et la face supérieure (17) ; ledit support (40) présentant une direction gauche-droite transversale à la face gauche (19) et à la face droite (20) tandis qu’elle est parallèle à la face avant (21 ), à la face arrière (22) et aux faces principales, une direction avant-arrière transversale à la face avant (21 ) et à la face arrière (22) tandis qu’elle est parallèle à la face (19) gauche, à la face droite (20) et aux faces principales, et une direction bas-haut transversale aux faces principales tandis qu’elle est parallèle aux faces latérales ; la lamelle couvre-objet (12) étant parallélépipédique et présentant deux faces principales, à savoir une face supérieure (23) et une face inférieure (24), et des faces latérales qui s’étendent de l’une à l’autre des faces principales, à savoir une face gauche (25), une face droite (26), une face avant (27) et une face arrière (28) ; la face inférieure (24) de la lamelle couvre-objet (12) étant en regard de la face supérieure (17) de la lame porte-objet (11 ) tandis que la face avant (27), la face arrière (28), la face gauche (25) et la face droite (26) de la lamelle couvre-objet (12) sont tournées vers et à distance de respectivement la face avant (21 ), la face arrière (22), la face gauche (19) et la face droite (20) de la lame porte-objet (11 ) ; chaque dit compartiment individuel (14) étant délimité suivant la direction bas-haut respectivement par la face supérieure (17) de la lame porte-objet (11 ) et par la 1 . Support for the analysis by microscopy of a liquid-based biological substance, such as animal semen, comprising a microscope slide (11), a microscope slide (12) and lines of glue (13) arranged between the microscope slide (11) and the microscope slide (12) to secure them to one another while peripherally delimiting individual compartments (14) for receiving a sample of said biological substance; the slide (11) being parallelepiped and having two main faces, namely an upper face (17) and a lower face (18), and side faces which extend from one to the other of the faces main ones, namely a left face (19), a right face (20), a front face (21) and a rear 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 rear face (22) which is itself greater than the distance between the lower face (18) and the upper face (17); said support (40) having a left-right direction transverse to the left face (19) and to the right face (20) while it is parallel to the front face (21), to the rear face (22) and to the main faces, a front-rear direction transverse to the front face (21) and to the rear face (22) while it is parallel to the left face (19), to the right face (20) and to the main faces, and a bottom-up direction transverse to the main faces while it is parallel to the side faces; the coverslip (12) being parallelepipedic and having two main faces, namely an upper face (23) and a lower face (24), and side faces which extend from one to the other of the faces main, 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) facing 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 specimen slide (11); each said individual compartment (14) being delimited in the bottom-top direction respectively by the upper face (17) of the slide (11) and by the
25 face inférieure (24) de la lamelle couvre-objet (12) et délimité périphériquement par lesdites lignes de colle (13), chaque dit compartiment individuel (14) s’ouvrant exclusivement d’un côté sur un puits (15) et d’un autre côté sur un évent (16), chaque puits (15) et chaque évent (16) étant délimité par lesdites lignes de colle (13), chaque puits (15) étant tourné vers l’une des face avant (21 ) et face arrière (22) de la lame porte-objet (11 ) et chaque évent (16) étant tourné vers l’autre des face avant (21 ) et face arrière (22) de la lame porte-objet (11 ) ; caractérisé en ce que la face supérieure (17) de la lame porte-objet (11 ) est à nu et à découvert sur au-moins une région marginale avant (41 ) et sur au- moins une région marginale arrière (42), la région marginale avant (41 ) longeant la face avant (21 ) de la lame porte-objet (11 ), la région marginale arrière (42) longeant la face arrière (22) de la lame porte-objet (11 ), chacune desdites région marginale avant (41 ) et région marginale arrière (42) ayant une extension suivant la direction avant-arrière au-moins égale à 2,5 mm et une extension suivant la direction gauche-droite au moins égale à l’écart entre les axes (38) respectifs des deux compartiments individuels (14) les plus éloignés l’un de l’autre et au plus égale à la distance entre la face gauche (19) et la face droite (20) de la lame porte-objet. 25 lower face (24) of the coverslip (12) and delimited peripherally by said lines of glue (13), each said individual compartment (14) opening exclusively on one side onto a well (15) and on another side on a vent (16), each well (15) and each vent (16) being delimited by said lines of glue (13), each well (15) being turned towards one of the front faces (21) and rear face (22) of the object slide (11) and each vent (16) facing the other of the front face (21) and rear face (22) of the object slide (11); characterized in that the upper face (17) of the slide (11) is bare and uncovered 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 microscope slide (11), the rear marginal region (42) running along the rear face (22) of the microscope slide (11), each of said regions front marginal (41) and rear marginal region (42) having an extension along the front-rear direction at least equal to 2.5 mm and an extension along the left-right direction at least equal to the distance between the axes ( 38) respectively of the two individual compartments (14) farthest 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 selon la revendication 1 , caractérisé en ce que pour l’une au moins de la région marginale avant (41 ) et de la région marginale arrière (42), l’extension suivant la direction gauche-droite est égale à la distance entre la face gauche (19) et la face droite (20) de la lame porte-objet (11 ). 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 in the left-right direction is equal to the distance between the left face (19) and the right face (20) of the specimen slide (11).
3. Support selon l'une quelconque des revendications 1 ou 2, caractérisé en ce que chaque puits (15) a une extension suivant la direction avant-arrière qui est inférieure à l’extension suivant la direction avant-arrière de chacune desdites région marginale avant (41 ) et région marginale arrière (42). 3. Support according to any one of claims 1 or 2, characterized in that each well (15) has an extension in the front-rear direction which is less than the extension in the front-rear direction of each of said marginal regions. front (41) and rear marginal region (42).
4. Support selon l'une quelconque des revendications 1 à 3, caractérisé en ce que la distance entre la face gauche (19) et la face droite (20) de la lame porte-objet (11 ) est comprise entre 74,8 mm et 75,2 mm, la distance entre la face avant (21 ) et la face arrière (22) de la lame porte-objet (11 ) est comprise entre 24,8 mm et 25,2 mm, la distance entre la face gauche (25) et la face droite (26) de la lamelle couvre-objet (12) est comprise entre 31 ,8 mm et 32,2 mm, et la distance entre la face avant (27) et la face arrière (28) de la lamelle couvre-objet (12) est comprise entre 16,8 mm et 16,2 mm. 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 (11) is between 74.8 mm and 75.2 mm, the distance between the front face (21) and the rear face (22) of the microscope slide (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 selon l'une quelconque des revendications 1 à 4, caractérisé en ce que tous les puits (15) sont tournés vers l’une des face avant (21 ) et face arrière (22) de la lame porte-objet (11) et tous les évents (16) sont tournés vers l’autre des face avant (21 ) et face arrière (22) de la lame porte- objet (11 ). 5. Support according to any one of claims 1 to 4, characterized in that all the wells (15) face one of the front face (21) and rear face (22) of the slide (11 ) and all the vents (16) are facing the other of the front face (21) and rear face (22) of the slide (11).
6. Système pour l’analyse d’une substance biologique à base liquide, comportant un support (40) selon l'une quelconque des revendications 1 à 5 et un microscope (50) comportant un objectif (51), une platine (52) configurée pour recevoir le support (40) dans une première position d’observation dans laquelle la face inférieure (18) de la lame porte-objet (11 ) est contre la platine6. 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) et un des compartiments individuels (14) est aligné avec l’objectif (51 ), un bâti sur lequel l’objectif (51 ) est monté dans au moins une position prédéterminée fixe et sur lequel la platine (52) est montée mobile en translation suivant la direction gauche-droite du support (40) afin de pouvoir aligner successivement les autres compartiments individuels (14) avec l’objectif (51 ), une plaque d’appui (53) s’étendant au moins partiellement en regard du support (40), et un organe de serrage (54) configuré pour solliciter la plaque d’appui(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 in the left-right direction of the support (40) in order to be able to successively align the other individual compartments (14) with the lens (51), a support plate (53) extending at least partially opposite of the support (40), and a clamp (54) configured to bias the backing plate
(53) et la platine (52) l’une vers l’autre de sorte que le support (40) est enserré entre la plaque d’appui (53) et la platine (52) ; la plaque d’appui (53) étant montée fixe sur le bâti et comportant une embase (55) et des doigts d’appui(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) saillant de l’embase (55) vers la platine (52) et dont les extrémités distales(56) projecting from the base (55) towards the plate (52) and whose distal ends
(57) sont au contact des régions marginales (41 , 42) exclusivement, avec au moins deux doigts d’appui (56) sur l’une des région marginale avant (41 ) et région marginale arrière (42) et au moins un doigt d’appui (56) sur l’autre des région marginale avant (41 ) et région marginale arrière (42) ; la platine (52) présentant des butées de positionnement (34, 35) pour immobiliser le support (40) par rapport à la platine (52) suivant la direction gauche-droite. (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 finger support (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. Système selon la revendication 6, caractérisé en ce que la plaque d’appui (53) comporte quatre doigts d’appui (56), dont deux doigts d’appui (56) au contact de la région marginale avant (41 ) et les deux autres doigts d’appui (56) au contact de la région marginale arrière (42). 7. System according to claim 6, characterized in that the bearing plate (53) comprises four bearing fingers (56), two of which bearing fingers (56) in contact with the front marginal region (41) and the other two support fingers (56) in contact with the rear marginal region (42).
8. Système selon l'une quelconque des revendications 6 ou 7, caractérisé en ce que l’un de la platine (52) et de la plaque d’appui (53) a des propriétés magnétiques et l’organe de serrage comporte des aimants (54) solidaires de l’autre de la platine (52) et de la plaque d’appui (53). 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) secured to the other of the plate (52) and the support plate (53).
9. Système selon l'une quelconque des revendications 6 à 8, caractérisé en ce que l’embase (55) présente une ouverture (58) au travers de laquelle l’objectif (51 ) pointe sur ledit compartiment individuel (14) aligné avec l’objectif (51 ) dans ladite première position d’observation, au-moins deux des doigts d’appui (56) qui sont au contact d’une même région marginale (41 , 42) étant disposés de part et d’autre de l’ouverture (58). 9. System according to any one of claims 6 to 8, characterized in that the base (55) has an opening (58) through which the lens (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. Système selon la revendication 9, caractérisé en ce que le bâti comporte une paroi de montage (60) et l’embase (55) comporte deux branches (61 ) saillant chacune transversalement de la paroi de montage (60), s’étendant en regard l’une de l’autre et délimitant l’ouverture (58), chaque branche (61 ) étant pourvue de deux doigts d’appui (56), dont un doigt (56) au contact de la région marginale avant (41) et un doigt (56) au contact de la région marginale arrière (42). 10. System according to claim 9, characterized in that the frame comprises a mounting wall (60) and the base (55) comprises two branches (61) each projecting transversely from the mounting wall (60), extending opposite each other and delimiting the opening (58), each branch (61) being provided with two support fingers (56), one finger (56) of which is in contact with the front marginal region (41 ) and a finger (56) in contact with the rear marginal region (42).
11 . Système selon la revendication 10, caractérisé en ce que l’organe de serrage comporte des aimants (54) logés dans lesdites branches (61 ). 11 . System according to Claim 10, characterized in that the clamping member comprises magnets (54) housed in the said branches (61).
12. Système selon l'une quelconque des revendications 6 à 11 , caractérisé en ce que l’une de la platine (52) et de la plaque d’appui (53) présente une rampe (63) et l’autre de la platine (52) et de la plaque d’appui (53) présente un bossage (64) configuré pour coopérer avec la rampe (63) au cours du mouvement de translation de la platine (52) suivant la direction gauche- droite de manière à écarter la platine (52) de la plaque d’appui (53), à l’encontre de la force exercée par l’organe de serrage (54), jusqu’à une position dans laquelle le support (40) est accessible pour pouvoir être retiré de la platine (52). 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 separate the plate (52) of the support plate (53), against the force exerted by the clamping member (54), to a position in which the support (40) is accessible in order to be able to be removed from the plate (52).
13. Système selon l'une quelconque des revendications 6 à 12, caractérisé en ce que la platine (52) est mécaniquement reliée au bâti par une 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
28 liaison pivot glissant comportant un guide cylindrique (65) longitudinalement orienté suivant la direction gauche-droite et un fût (66) monté à coulissement sur le guide (65), avec un jeu entre le fût (66) et le guide (65). 28 sliding pivot connection comprising a cylindrical guide (65) longitudinally oriented in the left-right direction and a barrel (66) mounted to slide on the guide (65), with a clearance between the barrel (66) and the guide (65).
14. Système selon l'une quelconque des revendications 6 à 13, caractérisé en ce que l’objectif (51 ) a une distance focale fixe. 14. System according to any one of claims 6 to 13, characterized in that the lens (51) has a fixed focal length.
15. Utilisation d’un système selon l'une quelconque des revendications 6 à 14, caractérisé en ce que la substance biologique à base liquide est de la semence animale et ladite analyse comprend une étape de comptage des spermatozoïdes. 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 comprises a sperm count step.
29 29
EP21851668.0A 2020-12-22 2021-12-13 Support for analysing by microscope a liquid-based biological substance and system comprising such a support and a microscope Pending EP4267302A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
FR2013942A FR3117896A1 (en) 2020-12-22 2020-12-22 Support for microscopic analysis of a liquid-based biological substance and system comprising such a support and a microscope
PCT/FR2021/052305 WO2022136766A1 (en) 2020-12-22 2021-12-13 Support for analysing by microscope a liquid-based biological substance and system comprising such a support and a microscope

Publications (1)

Publication Number Publication Date
EP4267302A1 true EP4267302A1 (en) 2023-11-01

Family

ID=74669105

Family Applications (1)

Application Number Title Priority Date Filing Date
EP21851668.0A Pending EP4267302A1 (en) 2020-12-22 2021-12-13 Support for analysing by microscope a liquid-based biological substance and system comprising such a support and a microscope

Country Status (5)

Country Link
US (1) US20240042433A1 (en)
EP (1) EP4267302A1 (en)
CN (1) CN116847931A (en)
FR (1) FR3117896A1 (en)
WO (1) WO2022136766A1 (en)

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US566849A (en) 1896-09-01 Cable hoisting and conveying apparatus
NL9500281A (en) 1995-02-15 1996-09-02 Jan Pieter Willem Vermeiden Counting chamber for biological research as well as a method for the production of such a counting chamber.
KR970706902A (en) * 1995-09-12 1997-12-01 로드릭 리차드 제이 DEVICE AND METHOD FOR DNA AMPLIFICATION AND ASSAY
US6569674B1 (en) * 1999-12-15 2003-05-27 Amersham Biosciences Ab Method and apparatus for performing biological reactions on a substrate surface
WO2011026136A1 (en) * 2009-08-31 2011-03-03 Life Technologies Corporation Low-volume sequencing system and method of use
CN105675495A (en) * 2014-11-17 2016-06-15 吕慧菁 Detection device having magnifying function

Also Published As

Publication number Publication date
US20240042433A1 (en) 2024-02-08
CN116847931A (en) 2023-10-03
FR3117896A1 (en) 2022-06-24
WO2022136766A1 (en) 2022-06-30

Similar Documents

Publication Publication Date Title
EP0107989B1 (en) Apparatus for mounting optical fibres in a ferrule
EP0959343A1 (en) Optical sensor utilizing an immunological reaction and a fluorescence marker
FR2823373A1 (en) Device and method for automatic cutting of a layer of a substrate, comprises a holder and cutting means including a cleaving plate and two side plates mounted symmetrically
EP0146867B1 (en) Variable optical attenuator for coupling optical fibres
FR2606166A1 (en) OPTICAL FIBER SUPPORT AND DEVICE COMPRISING THE SAME
EP3069185A1 (en) Three-dimensional focusing device and method for a microscope
EP1952088A1 (en) Method for scanning rim groove contour of spectacle frame
EP0402230A1 (en) Process and device for marking and cleaving mono-crystalline semiconductor wafers
BE897778A (en) BLADE FOR MICROSCOPIC EXAMINATION
EP4267302A1 (en) Support for analysing by microscope a liquid-based biological substance and system comprising such a support and a microscope
WO2007068811A1 (en) Housing for assaying a biological sample by immunochromatography
FR2737779A1 (en) HIGH SPATIAL ELLIPSOMETER DEVICE
EP0724134A1 (en) Device for measuring the length and method for adjusting the said device
EP0733226B1 (en) Observation instrument orientation detecting system
FR2569835A1 (en) HEAD FOR MEASURING DIAMETERS OF CYLINDRICAL PARTS
EP0096608B1 (en) Optical fibre collimator, application to the realisation of optical switches
EP3317017B1 (en) Device for the analysis of a liquid sample comprising a blocking and unblocking element
EP4016054A1 (en) Optical component for an atr imaging device
FR3067313B1 (en) END STICK FOR MOTOR VEHICLE WIPER BRUSH
EP0176397B1 (en) Apparatus for orienting an optical eleement
WO2007045734A1 (en) Appliance for tactile sensing a spectacle frame and related grinding machine
FR2823859A1 (en) Support for positioning and holding of optical fibers comprises first part for holding non-stripped part of fiber and second part for holding stripped part of fiber for connection to optical device
EP4165454A1 (en) Optical system with guide rail
WO2005038392A2 (en) Optical device for determining at least one dimensional characteristic of a drop of liquid
EP2813876B1 (en) Mounting intended in particular for systems such as lenses for microscopes and/or sensors

Legal Events

Date Code Title Description
STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: UNKNOWN

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE INTERNATIONAL PUBLICATION HAS BEEN MADE

PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: REQUEST FOR EXAMINATION WAS MADE

17P Request for examination filed

Effective date: 20230704

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR

DAV Request for validation of the european patent (deleted)
DAX Request for extension of the european patent (deleted)