CN104614515B - The using method of the microslide couveuse of detachable and airtight space - Google Patents

The using method of the microslide couveuse of detachable and airtight space Download PDF

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Publication number
CN104614515B
CN104614515B CN201510091593.1A CN201510091593A CN104614515B CN 104614515 B CN104614515 B CN 104614515B CN 201510091593 A CN201510091593 A CN 201510091593A CN 104614515 B CN104614515 B CN 104614515B
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microslide
frame
hatches
resilient clamp
upper cover
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CN104614515A (en
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邱奥望
刘庆淮
龚琦
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Jiangsu Province Hospital First Affiliated Hospital With Nanjing Medical University
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Jiangsu Province Hospital First Affiliated Hospital With Nanjing Medical University
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    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/5302Apparatus specially adapted for immunological test procedures
    • G01N33/5304Reaction vessels, e.g. agglutination plates
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/531Production of immunochemical test materials
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00Sampling; Preparing specimens for investigation
    • G01N1/28Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/30Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00Sampling; Preparing specimens for investigation
    • G01N1/28Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/30Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
    • G01N1/31Apparatus therefor

Abstract

The invention discloses a kind of using method of microslide couveuse of detachable and airtight space, couveuse comprises stainless steel and hatches frame, stainless steel is hatched frame and is pressed on microslide, is that bottom surface is formed and hatches cell jointly with the silica gel bed course below it by the coffin of centre with microslide; Separately be provided with and stainless steel hatched frame and silica gel bed course and be clamped in resilient clamp on microslide; Stainless steel is hatched rectangular wall in frame and is provided with step trough, ensures that camera incubata upper cover can cover stably on the step trough that stainless steel hatches rectangular wall in frame, and form airtight space hatch cell; Described camera incubata upper cover comprises camera incubata upper cover cover plate and camera incubata upper cover handle.The present invention is rational in infrastructure, only uses the antibody incubation liquid of trace, all histotomies be pasted onto on microslide just can be allowed to be fully immersed in antibody incubation liquid, both saved the experimental cost of scientific research, maintains again the stable of antibody incubation liquid composition.

Description

The using method of the microslide couveuse of detachable and airtight space
The application is application number: the divisional application of 201410009410.2, the applying date: 2014.1.9, the title microslide couveuse of airtight space " detachable and ".
Technical field
The present invention relates to a kind of microslide couveuse of detachable and airtight space.
Background technology
In the research carrying out life science, people often need to use high specificity, highly sensitive immunohistochemistry technology to position histiocytic predetermined substance and quantitatively.The ultimate principle of immunohistochemistry technology first utilizes the combination needing to identify antigen in immunologic known antibodies (first antibody) and tissues observed.Due to the known antibodies that completes and need identify that the combination of antigen is generally sightless on tissue and cell, certain label (as enzyme, fluorescein) is attached on first antibody by the method for needs mark, then with the fluorescence that histochemical method shows this label or sends at fluorescent microscope, lazer scan confocal microscope or ultrahigh resolution basis of microscopic observation fluorescein.By visual for the need identification antigen in research object tissue.Above-mentioned mention with mark method certain label (as enzyme, fluorescein) is attached on first antibody time, people often adopt the method having connected the second antibody of certain label and the specific binding of first antibody.
Positioning with time quantitative with immunohistochemistry technology to histiocytic predetermined substance, the structural form that during the method that people often adopt paster to dye keeps testing, histotomy is good the tandem relation of clearly each histotomy.The ultimate principle of the method for paster dyeing is on microtome after Cut tissue sheet, is pasted onto on microslide by the definite sequence of this histotomy by section immediately.The processing procedure of the histotomies such as the closing of follow-up histotomy, first antibody is hatched, washed, second antibody is hatched, wash, dewater again, transparent and mounting is all carry out under this histotomy is pasted onto the condition of microslide.
The closing of above-mentioned histotomy, wash, wash again, dewater, in transparent and mounting process, because the regarding liquid cost used is lower, in addition in washing process, the large usage quantity of cleansing solution.In order to obtain good experiment effect, people often adopt the method by the whole microslide being pasted with histotomy is placed in above-mentioned related liquid to process.But when carrying out first antibody and second antibody is hatched, because antibody price is higher, (price of monoclonal primary antibody 100 μ l general is at present about 3000 yuan, special monoclonal first antibody price can be higher), and when using, attenuable multiple again can not be very large.Therefore, antibody especially first antibody become the major part of immunohistochemical assay cost.So positioning with time quantitative with immunohistochemistry technology to histiocytic predetermined substance, the method by the whole microslide being pasted with histotomy is placed in antibody incubation liquid can not be adopted.
People are when paster method carries out immunohistochemical staining, and antibody incubation liquid is directly added in the method on the histotomy of microslide by employing usually.But when using this method, antibody incubation liquid has been added and can not preserved and diffuse out microslide loss, adding the antibody incubation liquid lacked on histotomy can, in the process of hatching, can not keep the histotomy on microslide to be immersed in all the time in antibody incubation liquid because of moisture evaporation in Incubating Solution.Sometimes, people also can be used in the method for to draw a frame around histotomy that microslide attaches with oily material, and add antibody incubation liquid in this oiliness picture frame, prevent the spilling of antibody incubation liquid.But though the method that the method is directly added in than by antibody incubation liquid on the histotomy of microslide attaching has improvements, but use the method, due to the poor ability that oiliness picture frame stops antibody incubation liquid excessive, can not ensure that histotomy is immersed in antibody incubation liquid all the time in whole antibody incubation process.And in antibody incubation, washing, dehydration, clearing process, these oily materials can lose because coming off and stop the effect that in antibody incubation process, antibody incubation liquid is excessive; When oily material occurring and coming off and adhere on tissue sections, final sections observation effect can be affected; And oily material, also can be molten in relevant experimental solutions in the processing procedure of histotomy, pollute experimental solutions and the recycling of experimental solutions that affects and the treatment effect of histotomy.
Summary of the invention
The object of the present invention is to provide a kind of rational in infrastructure, all experimental procedure energy convenience and high-efficiencies carry out, and can obtain the microslide couveuse of the detachable of good immunohistochemical staining experimental result and airtight space.
Technical solution of the present invention is:
A microslide couveuse for detachable and airtight space, it is characterized in that: comprise stainless steel and hatch frame, stainless steel is hatched frame and is pressed on microslide, is that bottom surface is formed and hatches cell jointly with the silica gel bed course below it by the coffin of centre with microslide; Described silica gel bed course has hatches the identical intermediate rectangular space of frame with stainless steel, and sticks to stainless steel and hatch on frame, contacts, prevent the leakage of Incubating Solution with microslide; The peripheral dimension that stainless steel hatches the peripheral dimension of frame and silica gel bed course is all identical with microslide peripheral dimension; Separately be provided with and stainless steel hatched frame and silica gel bed course and be clamped in resilient clamp on microslide; Described resilient clamp comprises resilient clamp root, the clamping part of resilient clamp and the clamp port of resilient clamp; Resilient clamp root connects resilient clamp clamping part, and provides the hand position of resilient clamp when forming microslide couveuse and the couveuse operation of dismounting microslide; Being clamped in by resilient clamp by the clamp port of resilient clamp hatches on frame and microslide, and the clamping part of resilient clamp will be hatched the formation that is clamped together of frame and microslide and be hatched cell; Stainless steel is hatched rectangular wall in frame and is provided with step trough, ensures that camera incubata upper cover can cover stably on the step trough that stainless steel hatches rectangular wall in frame, and form airtight space hatch cell; Described camera incubata upper cover comprises camera incubata upper cover cover plate and camera incubata upper cover handle, and camera incubata upper cover cover plate covers hatching in frame on rectangle step trough, and hatches frame, and what microslide formed airtight space jointly hatches cell; Camera incubata upper cover handle is placed in the upper surface of camera incubata upper cover, opens or cover camera incubata upper cover by this handle hand-held.
Using method is: stainless steel is hatched frame and silicagel pad and be laminated to and be pasted with on the microslide of histotomy, guarantees that all histotomies are all hatched bottom cell what hatch that frame and microslide formed; Select to hatch frame and any one side in microslide up and down overlapping four limits, hand-held resilient clamp root, by the clamp port of resilient clamp, aim at the structure of hatching frame and microslide and being stacked up and down, resilient clamp is pushed the structure of hatching frame and microslide and being stacked up and down, ensure that the clamping part of resilient clamp will hatch frame and microslide forcing together tightly, its excess-three limit is repeated above-mentioned action and is sandwiched by resilient clamp, guarantee to hatch frame and microslide close contact, what formation was not leaked hatches cell; To hatching in cell the Incubating Solution adding first antibody, ensure that being pasted onto the histotomy of hatching bottom cell can be immersed in after in the Incubating Solution of first antibody, hand-held camera incubata upper cover handle, camera incubata upper cover cover plate is covered on the step trough that stainless steel hatches rectangular wall in frame, what form airtight space hatches cell, carries out hatching of histotomy on request; After having hatched for the first time, hand-held camera incubata upper cover handle opens camera incubata upper cover cover plate, the Incubating Solution of sucking-off first antibody, take off the resilient clamp of hatching frame and microslide phase stack structure surrounding up and down, under microslide dismounting, hatch frame, carrying out washing treatment is carried out to the histotomy be pasted onto on microslide; Need to carry out second antibody when hatching, repeat above-mentioned action.
The present invention is rational in infrastructure, when carrying out histiocytic immunohistochemical staining, by repeatedly using resilient clamp to be pasted with hatching frame being pressed in tightly on the microslide of histotomy, and cover camera incubata upper cover, what form an airtight space hatches cell.When carrying out antibody incubation, only use the antibody incubation liquid of trace, the all histotomies be pasted onto on microslide just can be allowed to be fully immersed in antibody incubation liquid, to prevent in the process of hatching and change the volume of Incubating Solution and the concentration of first antibody because of the evaporation of moisture in Incubating Solution.Both save the experimental cost of scientific research, maintain again the stable of antibody incubation liquid composition.Meanwhile, hatching frame by repeatedly unloading, in turn ensure that microslide pre-treatment, organize paster, washing, dehydration, the experimental procedure such as transparent, mounting by the impact of hatching frame, and can not avoid the shortcoming using oily material picture frame around histotomy.All experimental procedure energy convenience and high-efficiencies are carried out, guarantees finally to obtain a good immunohistochemical Coloration experiment result.
Accompanying drawing explanation
Below in conjunction with drawings and Examples, the invention will be further described.
Fig. 1 is detachable and the camera incubata upper cover schematic perspective view of the microslide couveuse of airtight space.
Fig. 2 be detachable and the microslide couveuse of airtight space hatch frame schematic perspective view.
Fig. 3 is detachable and the microslide schematic perspective view of the microslide couveuse of airtight space.
Fig. 4 is detachable and the resilient clamp schematic perspective view of the microslide couveuse of airtight space.
Fig. 5 is detachable and the microslide couveuse longitudinal profile schematic diagram of airtight space.
In figure:
1. camera incubata upper cover handle: the upper surface being placed in camera incubata upper cover, opens or covers camera incubata upper cover by this handle hand-held.
2. camera incubata upper cover cover plate: cover hatching in frame on rectangle step trough, and hatches frame, and what microslide formed airtight space jointly hatches cell.
3. stainless steel hatches frame: be pressed on microslide, is that bottom surface is formed and hatches cell jointly with the silica gel bed course below it by the coffin of centre with microslide
4. stainless steel hatches the step trough of rectangular wall in frame: ensure that camera incubata upper cover can cover stably on the step trough that stainless steel hatches rectangular wall in frame, and form airtight space hatch cell.
5. stainless steel hatches the silica gel bed course of frame: have and hatch the identical intermediate rectangular space of frame with stainless steel, and sticks to stainless steel and hatch on frame, contacts, prevent the leakage of Incubating Solution with microslide.
6. microslide: carrying histotomy, and as hatching the bottom surface of cell.
7. resilient clamp root: connect resilient clamp clamping part, and the hand position of resilient clamp when forming microslide couveuse and the couveuse operation of dismounting microslide is provided.
8. the clamping part of resilient clamp: the formation that is clamped together of frame and microslide will be hatched and hatch cell.
9. the clamp port of resilient clamp: by this clamp port resilient clamp is clamped in and hatches on frame and microslide.
10. camera incubata upper cover.
11. resilient clamps.
12. airtight space hatch cell: form small confined space and deposit Incubating Solution, guarantee under the prerequisite of the Incubating Solution using trace, the histotomy be pasted onto on microslide can be made to be immersed in antibody incubation liquid all the time, also prevent the evaporation of moisture in antibody incubation liquid, maintain the stable of antibody concentration.
Embodiment
A microslide couveuse for detachable and airtight space, comprise stainless steel and hatch frame, stainless steel is hatched frame and is pressed on microslide, is that bottom surface is formed and hatches cell jointly with the silica gel bed course below it by the coffin of centre with microslide; Described silica gel bed course has hatches the identical intermediate rectangular space of frame with stainless steel, and sticks to stainless steel and hatch on frame, contacts, prevent the leakage of Incubating Solution with microslide; The peripheral dimension that stainless steel hatches the peripheral dimension of frame and silica gel bed course is all identical with microslide peripheral dimension; Separately be provided with and stainless steel hatched frame and silica gel bed course and be clamped in resilient clamp on microslide; Described resilient clamp comprises resilient clamp root, the clamping part of resilient clamp and the clamp port of resilient clamp; Resilient clamp root connects resilient clamp clamping part, and provides the hand position of resilient clamp when forming microslide couveuse and the couveuse operation of dismounting microslide; Being clamped in by resilient clamp by the clamp port of resilient clamp hatches on frame and microslide, and the clamping part of resilient clamp will be hatched the formation that is clamped together of frame and microslide and be hatched cell; Stainless steel is hatched rectangular wall in frame and is provided with step trough, ensures that camera incubata upper cover can cover stably on the step trough that stainless steel hatches rectangular wall in frame, and form airtight space hatch cell; Described camera incubata upper cover comprises camera incubata upper cover cover plate and camera incubata upper cover handle, and camera incubata upper cover cover plate covers hatching in frame on rectangle step trough, and hatches frame, and what microslide formed airtight space jointly hatches cell; Camera incubata upper cover handle is placed in the upper surface of camera incubata upper cover, opens or cover camera incubata upper cover by this handle hand-held.
The microslide selected is processed, to ensure that histotomy has good sticking effect on microslide according to immunohistochemical requirement.Choose the required tissue of experiment to cut into slices, and be attached on microslide.The histotomy be attached on microslide is implemented the Seal treatment before antibody incubation.After completing above-mentioned experimental procedure, stainless steel is hatched frame (3) and silica gel bed course (5) be pressed in be pasted with histotomy microslide (6) on, guarantee that all histotomies all hatch cell (12) bottom what hatch that frame and microslide formed.Select to hatch frame and any one side in microslide up and down overlapping four limits, hand-held resilient clamp root (7), the clamp port (9) of resilient clamp is aimed at the structure of hatching frame and microslide and being stacked up and down, resilient clamp is pushed the structure of hatching frame and microslide and being stacked up and down.Ensure that the clamping part (8) of resilient clamp will hatch frame and microslide forcing together tightly.Its excess-three limit is repeated above-mentioned action and is sandwiched by resilient clamp (11).Guarantee to hatch frame and microslide close contact, what formation was not leaked hatches cell.To hatching in cell the Incubating Solution added containing special first antibody, be sure of to be pasted onto after in the Incubating Solution that the histotomy of hatching bottom cell can be immersed in containing special first antibody, hand-held camera incubata upper cover handle (1), being covered by camera incubata upper cover cover plate (2) hatches on the step trough (4) of rectangular wall in frame at stainless steel, what form airtight space hatches cell, carries out hatching of histotomy on request.After first time hatched, hand-held camera incubata upper cover handle has opened camera incubata upper cover cover plate, and sucking-off, containing the Incubating Solution of special first antibody, is unloaded the resilient clamp of hatching frame and microslide phase stack structure surrounding up and down, under microslide is dismantled, hatched frame.Routinely carrying out washing treatment is carried out to the histotomy be pasted onto on microslide.Need to carry out second antibody when hatching, repeat above-mentioned action, what again form airtight space hatches cell, adds the Incubating Solution containing special second antibody, hatches histotomy.After having hatched, remove containing special second antibody Incubating Solution.Frame is hatched under dismounting.The histotomy be pasted onto on microslide is washed, dewaters, the aftertreatment such as transparent, mounting.After all process complete, examine under a microscope the immunohistochemical staining result of histotomy.

Claims (2)

1. the using method of the microslide couveuse of a detachable and airtight space, it is characterized in that: described couveuse comprises stainless steel and hatches frame, stainless steel is hatched frame and is pressed on microslide, is that bottom surface is formed and hatches cell jointly with the silica gel bed course below it by the coffin of centre with microslide; Described silica gel bed course has hatches the identical intermediate rectangular space of frame with stainless steel, and sticks to stainless steel and hatch on frame, contacts, prevent the leakage of Incubating Solution with microslide; The peripheral dimension that stainless steel hatches the peripheral dimension of frame and silica gel bed course is all identical with microslide peripheral dimension; Separately be provided with and stainless steel hatched frame and silica gel bed course and be clamped in resilient clamp on microslide; Described resilient clamp comprises resilient clamp root, the clamping part of resilient clamp and the clamp port of resilient clamp; Resilient clamp root connects resilient clamp clamping part, and provides the hand position of resilient clamp when forming microslide couveuse and the couveuse operation of dismounting microslide; Being clamped in by resilient clamp by the clamp port of resilient clamp hatches on frame and microslide, and the clamping part of resilient clamp will be hatched the formation that is clamped together of frame and microslide and be hatched cell; Stainless steel is hatched rectangular wall in frame and is provided with step trough, ensures that camera incubata upper cover can cover stably on the step trough that stainless steel hatches rectangular wall in frame, and form airtight space hatch cell; Described camera incubata upper cover comprises camera incubata upper cover cover plate and camera incubata upper cover handle, and camera incubata upper cover cover plate covers on the step trough of hatching rectangular wall in frame, and hatches frame, and what microslide formed airtight space jointly hatches cell; Camera incubata upper cover handle is placed in the upper surface of camera incubata upper cover, opens or cover camera incubata upper cover by this handle hand-held;
Choose the required tissue of experiment to cut into slices, and be attached on microslide; The histotomy be attached on microslide is implemented the Seal treatment before antibody incubation; After completing above-mentioned experimental procedure, stainless steel is hatched frame and silicagel pad and be laminated to and be pasted with on the microslide of histotomy, guarantee that all histotomies are all hatched bottom cell what hatch that frame and microslide formed; Select to hatch frame and any one side in microslide up and down overlapping four limits, hand-held resilient clamp root, the clamp port of resilient clamp is aimed at the structure of hatching frame and microslide and being stacked up and down, resilient clamp is pushed the structure of hatching frame and microslide and being stacked up and down; Ensure that the clamping part of resilient clamp will hatch frame and microslide forcing together tightly; Its excess-three limit is repeated above-mentioned action and is sandwiched by resilient clamp; Guarantee to hatch frame and microslide close contact, what formation was not leaked hatches cell; To hatching in cell the Incubating Solution added containing special first antibody, be sure of to be pasted onto after in the Incubating Solution that the histotomy of hatching bottom cell can be immersed in containing special first antibody, hand-held camera incubata upper cover handle, camera incubata upper cover cover plate is covered on the step trough that stainless steel hatches rectangular wall in frame, what form airtight space hatches cell, carries out hatching of histotomy on request; After first time hatched, hand-held camera incubata upper cover handle has opened camera incubata upper cover cover plate, and sucking-off, containing the Incubating Solution of special first antibody, is unloaded the resilient clamp of hatching frame and microslide phase stack structure surrounding up and down, under microslide is dismantled, hatched frame; Routinely carrying out washing treatment is carried out to the histotomy be pasted onto on microslide; Need to carry out second antibody when hatching, repeat above-mentioned action, what again form airtight space hatches cell, adds the Incubating Solution containing special second antibody, hatches histotomy; After having hatched, remove containing special second antibody Incubating Solution; Frame is hatched under dismounting; The histotomy be pasted onto on microslide is washed, dewaters, transparent, mounting aftertreatment; After all process complete, examine under a microscope the immunohistochemical staining result of histotomy.
2. the using method of the microslide couveuse of detachable and airtight space according to claim 1, it is characterized in that: airtight space hatch cell: form small confined space and deposit Incubating Solution, guarantee under the prerequisite of the Incubating Solution using trace, the histotomy be pasted onto on microslide can be made to be immersed in antibody incubation liquid all the time, also prevent the evaporation of moisture in antibody incubation liquid, maintain the stable of antibody concentration.
CN201510091593.1A 2014-01-09 2014-01-09 The using method of the microslide couveuse of detachable and airtight space Expired - Fee Related CN104614515B (en)

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