CN105547794A - Immune staining machine - Google Patents
Immune staining machine Download PDFInfo
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- CN105547794A CN105547794A CN201610090715.XA CN201610090715A CN105547794A CN 105547794 A CN105547794 A CN 105547794A CN 201610090715 A CN201610090715 A CN 201610090715A CN 105547794 A CN105547794 A CN 105547794A
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- 238000010186 staining Methods 0.000 title abstract description 9
- 238000010438 heat treatment Methods 0.000 claims abstract description 11
- 239000011521 glass Substances 0.000 claims description 59
- 230000001575 pathological effect Effects 0.000 claims description 34
- 238000012744 immunostaining Methods 0.000 claims description 24
- 239000012530 fluid Substances 0.000 claims description 17
- 238000006073 displacement reaction Methods 0.000 claims description 11
- 229920003023 plastic Polymers 0.000 claims description 3
- 239000000427 antigen Substances 0.000 abstract description 11
- 102000036639 antigens Human genes 0.000 abstract description 11
- 108091007433 antigens Proteins 0.000 abstract description 11
- 238000000034 method Methods 0.000 description 14
- 239000007788 liquid Substances 0.000 description 8
- 238000004043 dyeing Methods 0.000 description 5
- 238000005516 engineering process Methods 0.000 description 3
- 238000000684 flow cytometry Methods 0.000 description 3
- 238000010166 immunofluorescence Methods 0.000 description 3
- 238000003825 pressing Methods 0.000 description 3
- 230000001737 promoting effect Effects 0.000 description 3
- 101710098119 Chaperonin GroEL 2 Proteins 0.000 description 2
- 238000005213 imbibition Methods 0.000 description 2
- 238000009434 installation Methods 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 101000894412 Mycolicibacterium paratuberculosis (strain ATCC BAA-968 / K-10) Bacterioferritin Proteins 0.000 description 1
- 101000588258 Taenia solium Paramyosin Proteins 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000027455 binding Effects 0.000 description 1
- 238000004040 coloring Methods 0.000 description 1
- 230000002380 cytological effect Effects 0.000 description 1
- 238000013016 damping Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000018044 dehydration Effects 0.000 description 1
- 238000006297 dehydration reaction Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000003053 immunization Effects 0.000 description 1
- 238000002649 immunization Methods 0.000 description 1
- 238000003125 immunofluorescent labeling Methods 0.000 description 1
- 238000011532 immunohistochemical staining Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000009870 specific binding Effects 0.000 description 1
- 238000007447 staining method Methods 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
- G01N1/31—Apparatus therefor
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Sampling And Sample Adjustment (AREA)
Abstract
The invention relates to an immune staining machine. The immune staining machine comprises a frame and supporting device, an immune staining device, an image collecting device, a heating device and a control system. Compared with the prior art, the immune staining machine has the advantages that multiple times of immune staining are performed on the same tissue slice, immune staining and image collecting can be performed alternatively on the same tissue slice, and multiple antigen expression conditions of the same cell are easily researched.
Description
Technical field
The invention belongs to checkout equipment technical field, what be specifically related to is a kind of immunostaining machine be applied in Hospital Pathological Department histotomy.
Background technology
Immunostaining is the principle utilizing antigen and antibody specific binding, detect the expression of specific protein (antigen), be find one of body method that changes of function is the most frequently used under physiology or pathological state, the most frequently used is immunohistochemical staining and immunofluorescence dyeing.In experimentation, often need research at least two kinds of antigen/protein expression in same cell or tissue sample, so that their respective location clear and definite or coexpression situation.The method the most often adopted is at present flow cytometry, but the cell concentration that this technology needs is larger, and be used for Flow cytometry poor effect through fixing and paraffin-embedded tissue, accurately can not show various antigen at tissue and cell natural distribution and status in vivo.If tissue specimen amount few (as cytological samples obtained by fine-needle aspiration) or the interior cell concentration to be detected of tissue are seldom, be then difficult to carry out Flow cytometry.By carrying out repeatedly to same sample the expression that immunostaining studies plurality of antigens on tissue sections, then can address this problem.Such as, after histotomy being carried out to the immunofluorescence dyeing of antigen A, due to being combined into and unstable Non-covalent binding of antigen and antibody, tissue can be put into damping fluid, suitable heating, just can make to separate to the antibody that antigen A combines and corresponding developer, can carry out again the immunofluorescence dyeing of antigen B, adopting uses the same method can also carry out the dyeing of antigens c, antigen D.There is the Multiple immunizations staining technique for histotomy at present, namely same histotomy is once dyeed, research plurality of antigens is expressed, as a kind of based on the Double-immunofluorescent labeling method (CN201410099873.2) of same Species origin first antibody, the shortcoming of this method is, in specific implementation process, the antigen carrying out marking is needed to be not only often two kinds, it is in most cases 5-10 kind, even reach more than 20 to plant, this multiple staining method can not be satisfied the demand, and needs repeatedly to dye.Certainly, multiple staining can be combined with repeatedly colouring method, can increase work efficiency.In addition, in repeatedly dyeing course, take pictures after needing to dye to histotomy, compared by stained photographs repeatedly, just can understand which antigen same cell expresses actually, thus, the device provided also will be convenient to take pictures under the microscope at every turn.Applicant discloses a kind of slide glass folder (201520587855.9) for carrying out repeatedly immunostaining to histotomy, a kind of slide glass folder for carrying out repeatedly immunostaining to histotomy being convenient to craft or machine full-automatic operation is provided, increasing and the not easily deficiency such as standardization to overcome labor capacity that manual operations brings, being necessary to provide a kind of full-automatic immunostainer possessing repeatedly immunostaining and image collecting function.
Summary of the invention
The object of the present invention is to provide a kind of immunostaining machine, its technical matters to be solved is: adopt existing immunostainer cannot carry out repeatedly immunostaining to same histotomy, cannot carry out immunostaining alternately and image acquisition to same histotomy.
For solving the problems of the technologies described above, the invention provides a kind of immunostaining machine, it comprises: framework and bracing or strutting arrangement, immunostaining device, image collecting device, heating arrangement and control system.
Described framework and bracing or strutting arrangement, by four big frame erection bolts and corresponding big frame mounting nuts, upper cover plate, lower cover, left cover, right cover plate, front shroud, back shroud and slide glass are pressed from both sides mounting sheet to be installed together, formed and be divided into upper and lower two-part rectangular box-like structure with slide glass folder mounting sheet.Described slide glass folder mounting sheet, in " H " shape, comprises a crossbeam and two arms.
Described immunostaining device, comprises micropipettor, pipettor moves carrier, upper hydraulic cylinder, longitudinal hydraulic cylinder, pipettor hydraulic cylinder, middle hydraulic cylinder, lower hydraulic cylinder, pipette slide glass, slide glass folder and vacuum pump, framework upper upper rail and framework upper lower guideway and cylindrical carrier.Cylindrical carrier to be arranged on above slide glass folder mounting sheet crossbeam position on the right side, for storing carrier fluid.Upper cover plate being opened with direction from left to right the window of a bar shaped, moving carrier at framework upper upper rail and framework upper lower guideway for installing pipettor.Described upper hydraulic cylinder is arranged on upper cover plate from left to right, moves carrier left and right displacement for promoting pipettor; Longitudinal hydraulic cylinder is arranged on upper cover plate, from the top down for promoting the upper and lower displacement of pipettor mobile vehicle; Pipettor hydraulic cylinder is arranged on mobile vehicle, can promote micropipettor and get liquid push rod, realizes micropipettor and gets liquid and blow liquid function; Near the position in left side below the crossbeam that middle hydraulic cylinder is arranged on slide glass folder mounting sheet from left to right, for promote pipette slide glass to slide glass folder mounting sheet institute lay slide glass press from both sides immediately below and reset accordingly.Described slide glass folder is arranged on the left side of cylindrical carrier on slide glass folder mounting sheet.It is in the same size that 1-20 opens slide glass folder, and the reserved installation position pressing from both sides mounting sheet with slide glass coincide, and all slide glasses press from both sides with equidirectional in the neat horizontal arranged in parallel of wire.Each hydraulic cylinder described all installs independently displacement transducer.
Described slide glass folder is as patent (a kind of slide glass folder for carrying out repeatedly immunostaining to histotomy: 201520587855.9), be made up of colourless transparent plastic, upper slice be sieve-like, be that there is at least one fluid hole at the edge of bottom sheet with a kind of difference of pressing from both sides for the slide glass carrying out repeatedly immunostaining to histotomy, this fluid hole communicates with the gap between bottom sheet with upper slice, to make slice the liquid between bottom sheet can be sopped up by this hole, upper slice and bottom sheet between carry histotomy.
Described image collecting device, comprises lower hydraulic cylinder, framework bottom lower guideway, framework bottom upper rail, digital pathological section scanning system mobile vehicle and digital pathological section scanning system.Described framework bottom lower guideway, framework bottom upper rail are arranged on lower cover with direction from left to right, described digital pathological section scanning system mobile vehicle is arranged on framework bottom lower guideway and framework bottom upper rail, described digital pathological section scanning system is arranged on digital pathological section scanning system mobile vehicle, described lower hydraulic cylinder is arranged on the left side of digital pathological section scanning system mobile vehicle, arrives slide glass folder position for promoting digital pathological section scanning system and resets accordingly.The difference of described digital pathological section scanning system and conventional digital pathological section scanning system is: the microscope adopted is inverted microscope, can provide at least white, red, green and blue four kinds of light sources.
Described master controller obtains the displacement transducer data of each hydraulic cylinder, and the accurate control by regulating corresponding hydraulic cylinder solenoid valve to realize each hydraulic cylinder;
Described master controller sends instruction after digital pathological section scanning system being moved to slide glass folder position and starts scanning to scanner, the data of scanning transfer to PC process, after a scan sample completes, scanner is moved to next sample position by master controller, and again send an instruction scanner, make it restart scanning.
Described master controller obtains temperature sensor data and realizes the control to sample environment temperature by adjustment heating arrangement.Described heating arrangement is arranged in lower plate, and temperature sensor is arranged in framework, and the temperature in framework is controlled at 37 DEG C (operating ambient temperatures must lower than 37 DEG C).
Principle of work and process:
Operationally, the slide glass carrying histotomy (paraffin section need through dehydration) is folded up the predeterminated position being placed in slide glass folder mounting sheet, carrier fluid is filled in cylindrical carrier; According to the carrier fluid position of intending drawing, upper Driven by Hydraulic Cylinder pipettor moves carrier and makes micropipettor displacement in left-right direction, after above arrival cylindrical carrier, longitudinal Hydraulic Cylinder pipettor mobile vehicle makes micropipettor to bottom offset, the suction nozzle of pipettor is declined and arrives carrier fluid, pipettor hydraulic cylinder is pulled upwardly micropipettor and gets liquid push rod, realizes micropipettor and gets liquid function; Then make micropipettor to top offset by longitudinal Driven by Hydraulic Cylinder pipettor mobile vehicle, the suction nozzle of pipettor is made to rise and leave carrier fluid, move carrier by upper Hydraulic Cylinder pipettor and make micropipettor displacement to the right, when moving to the slide glass folder position of intending filling carrier fluid, longitudinal Hydraulic Cylinder pipettor moves carrier and micropipettor is moved to downwards press from both sides close to slide glass, push away micropipettor downwards by pipettor hydraulic cylinder and get liquid push rod, liquid is filled on histotomy accurately.After the certain hour of setting, immediately below middle Hydraulic Cylinder pipette slide glass to slide glass folder, vacuum pump is opened, and suck the carrier fluid in slide glass folder, step that then can be similar carries out the filling of lower a kind of carrier fluid.Carry out image acquisition if intend, lower Driven by Hydraulic Cylinder digital pathological section scanning system presss from both sides position to slide glass, and lower Driven by Hydraulic Cylinder digital pathological section scanning system resets after scanning completes.After completing image acquisition, then carry out next step operation.In whole process, master controller accurately controls corresponding hydraulic cylinder solenoid valve by obtaining the displacement transducer data of each hydraulic cylinder; Accurate control figure pathological section scanning system moves to slide glass folder position and corresponding scanning process, and the data of scanning transfer to PC process; Obtain temperature sensor data and realize the control to sample environment temperature by adjustment heating arrangement.
The present invention's beneficial effect is compared with prior art: can carry out repeatedly immunostaining to same histotomy; Immunostaining alternately and image acquisition can be carried out to same histotomy.
Accompanying drawing illustrates:
Fig. 1 is positive sectional structure chart of the present invention;
Fig. 2 is left side view of the present invention;
Fig. 3 is right side view of the present invention;
Fig. 4 is slide glass of the present invention folder mounting sheet planform figure;
Fig. 5 is slide glass clamping structure figure of the present invention;
Fig. 6 is each hydraulic cylinder mounting means figure of the present invention;
Fig. 7 is control structure module schematic diagram of the present invention.
In figure: 1, big frame erection bolt; 2, big frame mounting nuts; 3, upper hydraulic cylinder; 4 ~ 7, upper hydraulic cylinder mounting screw; 8, framework upper upper rail; 9, framework upper upper rail mounting screw; 10 ~ 13, longitudinal hydraulic cylinder mounting screw; 14 ~ 17, longitudinal hydraulic cylinder mounting screw; 18, longitudinal hydraulic cylinder; 19, mobile carrier; 20, pipettor mobile vehicle; 21, framework upper lower guideway; 22, big frame mounting nuts; 23, big frame erection bolt; 24, pipettor hydraulic cylinder; 25, pipettor hydraulic cylinder mounting screw; 26, micropipettor; 27, pipettor mounting bar; 28, pipettor mounting screw; 29, carrier fluid and cylindrical carrier; 30, right cover plate; 31, lower cover; 32, heating arrangement; 33, framework bottom lower guideway; 34, framework bottom upper rail; 35, framework lower guide mounting screw; 36, digital pathological section scanning system mobile vehicle; 37 ~ 38, digital pathological section scanning system mounting screw; 39, digital pathological section scanning system; 40, vacuum pump mounting bracket; 41, vacuum pump; 42, middle hydraulic cylinder leg; 43, lower hydraulic cylinder; 44, middle hydraulic cylinder lateral frame; 45, middle hydraulic cylinder; 46, slide glass folder mounting sheet; 47, imbibition house steward; 48, middle hydraulic cylinder mounting screw; 49, pipette slide glass; 50, imbibition rubber tube; 51, left cover; 52, upper cover plate; 53, slide glass folder; 54, upper slice; 55, bottom sheet; 56, fluid hole; 57, histotomy.
Embodiment
Below in conjunction with accompanying drawing, embodiments of the present invention are described further:
Framework of the present invention and bracing or strutting arrangement, by four big frame erection bolts 1 and corresponding big frame mounting nuts 2, upper cover plate 52, lower cover 31, left cover 51, right cover plate 30, front shroud, back shroud and slide glass are pressed from both sides mounting sheet 46 to be installed together, formed and be divided into upper and lower two-part rectangular box-like structure with slide glass folder mounting sheet 46.Described slide glass folder mounting sheet 46, in " H " shape, comprises a crossbeam and two arms.
Described immunostaining device, comprises micropipettor 26, pipettor moves carrier 19, upper hydraulic cylinder 3, longitudinal hydraulic cylinder 18, pipettor hydraulic cylinder 24, middle hydraulic cylinder 45, lower hydraulic cylinder 43, pipette slide glass 49, slide glass folder 53 and vacuum pump 41, framework upper upper rail 8 and framework upper lower guideway 21 and cylindrical carrier 29.Cylindrical carrier 29 to be arranged on above slide glass folder mounting sheet 46 crossbeam position on the right side.Upper cover plate 52 being opened with direction from left to right the window of a bar shaped, moving carrier 19 at framework upper upper rail 8 and framework upper lower guideway 21 for installing pipettor.Described upper hydraulic cylinder 3 is arranged on upper cover plate 52 from left to right; Longitudinal hydraulic cylinder 18 is arranged on upper cover plate 52 from the top down; Pipettor hydraulic cylinder 24 is arranged on pipettor mobile vehicle 20; Near the position in left side below the crossbeam that middle hydraulic cylinder 45 is arranged on slide glass folder mounting sheet 46 from left to right; The left side of cylindrical carrier 29 on the crossbeam that described slide glass folder 53 is arranged on slide glass folder mounting sheet 46; 20 slide glasses press from both sides 53 in the same size, and the reserved installation position pressing from both sides mounting sheet 46 with slide glass coincide, all slide glass folders 53 with equidirectional in the neat horizontal arranged in parallel of wire; Each hydraulic cylinder described all installs independently displacement transducer.
Described slide glass folder 53, is made up of colourless transparent plastic, upper slice 54 be sieve-like, bottom sheet 55 only has a fluid hole 56 at edge, this fluid hole communicates with upper slice gap between 54 with bottom sheet 55, upper slice carry histotomy 57 between 54 and bottom sheet 55.
Described image collecting device, comprises lower hydraulic cylinder 43, framework bottom lower guideway 33, framework bottom upper rail 34, digital pathological section scanning system mobile vehicle 36 and digital pathological section scanning system 39.Described framework bottom lower guideway 33, framework bottom upper rail 34 to be arranged on lower cover 31 with direction from left to right, described digital pathological section scanning system mobile vehicle 36 is arranged on framework bottom lower guideway 33, framework bottom upper rail 34, described digital pathological section scanning system 39 is arranged on digital pathological section scanning system mobile vehicle 36, and lower hydraulic cylinder 43 is arranged on the left side of digital pathological section scanning system mobile vehicle 36.The microscope that described digital pathological section scanning system 39 adopts is inverted microscope, can provide white, red, green and blue four kinds of light sources.
Described master controller obtains the displacement transducer data of each hydraulic cylinder and accurately controls each hydraulic cylinder, and the data that digital pathological section scanning system 39 obtains are transferred to PC process; Described master controller obtains temperature sensor data and realizes the control to sample environment temperature by adjustment heating arrangement 32.Described heating arrangement 32 is arranged on lower cover 31, and temperature sensor is arranged in framework, makes the temperature in framework control at 37 DEG C; The operating ambient temperature of described a kind of immunostaining machine is room temperature.
Claims (3)
1. the invention provides a kind of immunostaining machine, it comprises: framework and bracing or strutting arrangement, immunostaining device, image collecting device, heating arrangement and control system;
Described framework and bracing or strutting arrangement, by four big frame erection bolts and corresponding big frame mounting nuts, upper cover plate, lower cover, left cover, right cover plate, front shroud, back shroud and slide glass are pressed from both sides mounting sheet to be installed together, formed and be divided into upper and lower two-part rectangular box-like structure with slide glass folder mounting sheet;
Described immunostaining device, comprise micropipettor, pipettor moves carrier, upper hydraulic cylinder, longitudinal hydraulic cylinder, pipettor hydraulic cylinder, middle hydraulic cylinder, lower hydraulic cylinder, pipette slide glass, slide glass folder and vacuum pump, framework upper upper rail and framework upper lower guideway and cylindrical carrier, cylindrical carrier to be arranged on above slide glass folder mounting sheet crossbeam position on the right side, described pipettor moves carrier and is arranged on framework upper upper rail and framework upper lower guideway, described upper hydraulic cylinder is arranged on upper cover plate from left to right, longitudinal hydraulic cylinder is arranged on upper cover plate from the top down, pipettor hydraulic cylinder is arranged on mobile vehicle, near the position in left side below the crossbeam that middle hydraulic cylinder is arranged on slide glass folder mounting sheet from left to right, described slide glass folder is arranged on the left side of cylindrical carrier on slide glass folder mounting sheet, all slide glasses press from both sides with equidirectional in the neat horizontal arranged in parallel of wire, each hydraulic cylinder described all installs independently displacement transducer,
Described image collecting device, comprise lower hydraulic cylinder, framework bottom lower guideway, framework bottom upper rail, digital pathological section scanning system mobile vehicle and digital pathological section scanning system, described framework bottom lower guideway, framework bottom upper rail is arranged on lower cover with direction from left to right, described digital pathological section scanning system mobile vehicle is arranged on framework bottom lower guideway and framework bottom upper rail, described digital pathological section scanning system is arranged on digital pathological section scanning system mobile vehicle, described lower hydraulic cylinder is arranged on the left side of digital pathological section scanning system mobile vehicle,
Described master controller obtains the displacement transducer data of each hydraulic cylinder and each hydraulic cylinder of accuracy controlling, obtains the data of digital pathological section scanning system collection and transfers to PC, obtains temperature sensor data and also regulates and controls heating arrangement.
2. a kind of immunostaining machine according to claim 1, is characterized in that: described slide glass folder is made up of colourless transparent plastic, upper slice be sieve-like, there is at least one fluid hole at the edge of bottom sheet, and this fluid hole communicates with the gap between bottom sheet with upper slice.
3. a kind of immunostaining machine according to claim 1, is characterized in that: the microscope that described digital pathological section scanning system adopts is inverted microscope, can provide at least white, red, green and blue four kinds of light sources.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610090715.XA CN105547794B (en) | 2016-02-18 | 2016-02-18 | A kind of immunostaining machine |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610090715.XA CN105547794B (en) | 2016-02-18 | 2016-02-18 | A kind of immunostaining machine |
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| CN105547794A true CN105547794A (en) | 2016-05-04 |
| CN105547794B CN105547794B (en) | 2018-12-04 |
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| CN201610090715.XA Expired - Fee Related CN105547794B (en) | 2016-02-18 | 2016-02-18 | A kind of immunostaining machine |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106289923A (en) * | 2016-09-29 | 2017-01-04 | 南昌德漫多科技有限公司 | A kind of device for same tissue slice being carried out repeatedly immunostaining |
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Effective date of registration: 20230420 Address after: 550081 area B, Jinhua garden south, Guanshan North Road, Jinhua Garden Street, guanshanhu District, Guiyang City, Guizhou Province Patentee after: Guiyang demandoo Medical Technology Co.,Ltd. Address before: 330001 Room 202, Unit 1, Residential Building, No. 39, Vanke Runyuan, Qingyunpu District, Nanchang City, Jiangxi Province Patentee before: NANCHANG DEMANDUO TECHNOLOGY Co.,Ltd. |
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