CN105021440A - Endocrine cell rapid dyeing method - Google Patents
Endocrine cell rapid dyeing method Download PDFInfo
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- CN105021440A CN105021440A CN201510474195.8A CN201510474195A CN105021440A CN 105021440 A CN105021440 A CN 105021440A CN 201510474195 A CN201510474195 A CN 201510474195A CN 105021440 A CN105021440 A CN 105021440A
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Abstract
The invention discloses an endocrine cell rapid dyeing method. The method includes the following steps that a tissue slice is placed in a dyeing rack and sequentially placed in dimethylbenzene I and dimethylbenzene II for 8 min respectively, after rehydration and distilled water cleaning, the slice is placed in a dye jar containing silver liquor, dyeing in dark is carried out for 3 hours, the slice is quickly washed twice with distilled water after the dye jar is cooled, and the silver liquor around the dye jar is wiped away through filter paper; then, the slice is reduced in reduction liquid for 1 hour, quickly washed twice with distilled water and processed in sodium thiosulfate for 2 minutes, the liquid is poured, and the liquid around the dye jar is wiped away through filter paper; the slice is placed in newly prepared silver liquor and quickly washed twice with distilled water after dyeing in dark, and the silver liquor around the dye jar is wiped away through filter paper; the slice is placed in newly prepared reduction liquid and quickly washed twice with distilled water after reduction is carried out for 1 minute, the tissue slice is placed in the dyeing rack again and dehydrated after redundant water is removed, and then transparent disposal is conducted. The method is simple in process, convenient to operate, good in dyeing effect and short in coloring time.
Description
Technical field
The present invention relates to cell dyeing field, be specifically related to a kind of rapid dyeing method of endocrine cell.
Background technology
The important step that smear staining film-making is medical test and biological study is carried out to cell or bacterium, has three kinds of methods to carry out smear, dyeing, film-making to cell or bacterium at present: one is traditional manual operation smear, dyeing, film-making; Another kind utilizes instrument and equipment automatic smear, and then take traditional manual operation dyeing, film-making; The third utilizes the automatic film-making of instrument and equipment.First method manual operation smear, dyeing, film-making, its instrument and equipment needs less, it is little to invest, but there is following defect:
1, smear thickness relies on the working experience of operating personnel, is difficult to control.
2, when for cytology film-making, the impurity in sample such as blood, mucus, bacterium, downright bad degenerated cell etc. cannot remove.
3, the experience with operating personnel drips dyeing liquor, and randomness is comparatively large, can not be quantitative, the poor repeatability of Color.
4, dyeing time, temperature can only guestimate, can not guarantee Color.
Second method utilizes instrument and equipment automatic smear, and then take the course of work of traditional manual operation stained preparation to be: utilize negative pressure to collect and sample is after treatment drawn into and is provided with in the filtration unit of filtering membrane, liquid part in sample is excluded by filtering membrane, visible component is distributed on filtering membrane, then utilize equipment by the visible component on filtering membrane, namely cell is transferred on slide, this completes automatic smear, utilize manually-operated method to carry out dyeing and film-making afterwards.
Its shortcoming is: 1, be only applicable to sectioning cells, can not be used for bacterium film-making.Reason is that bacterium is too little, cannot be shifted on slide after being collected by filtering membrane; 2, there is a process extruded when being transferred on slide by the cell that filtering membrane is collected, easily causing cell rupture or distortion, affecting production effect; 3, the experience with operating personnel drips dyeing liquor, and randomness is comparatively large, can not be quantitative, the poor repeatability of Color.4, dyeing time, temperature can only guestimate, can not guarantee Color.
The third method utilizes the automatic film-making of instrument and equipment.There is cytology automatic smear, the instrument and equipment of dyeing function only has the U.S. to produce at present, its type has Auto-Cyte PREP instrument etc., its principle of work is: sample is carried out gradient separations, centrifugal packed cells sample, being transferred to by sample is provided with in the special device of slide, by cell natural subsidence on slide, utilize the coating of sticking on slide to be fixed by cell, then automatic staining completes film-making process.Utilize this equipment production effect good, but automatic benchmarking originally can not carry out dilution process etc.; The cell thickness of film-making can not control; Function is few, can only complete a kind of colouring method of pap staining; Complicated operation, apparatus expensive, the domestic quotation of a set of the said equipment is more than 1,000,000 yuan, and consumables price, cost of use is high.
Summary of the invention
For solving the problem, the invention provides a kind of rapid dyeing method of endocrine cell, technique is simple, and easy to operate, Color is good, and colouring required time is short.
For achieving the above object, the technical scheme that the present invention takes is:
A rapid dyeing method for endocrine cell, comprises the steps:
Step 1, histotomy is put into staining rack be placed in 100% dimethylbenzene I, each 8min of 100% dimethylbenzene II successively;
Step 2, by the histotomy of step 1 gained by concentration be 100% dimethylbenzene I 5min, concentration to be 100% dimethylbenzene II 5min concentration be 95% alcohol 5min, concentration be 85% alcohol 5min, concentration be after the gradient of the alcohol 5min of 75% carries out rehydration, be placed in distilled water and clean 5min, then wash once with distilled water is anxious;
Step 3, the histotomy of step 2 gained is placed in the dye vat of the silvering solution being equipped with 60 DEG C, 60 DEG C of constant temperature lucifuges dyeing 3h, after dye vat cooling, distilled water is anxious washes 2 times, wipes silvering solution around dye vat with filter paper;
Step 4, the histotomy of step 3 gained put into the reducing solution of 45 DEG C, after 45 DEG C of constant temperature reduction 1h, distilled water is anxious washes 2 times;
Step 5, by reduction after histotomy put into concentration be 5% sodium thiosulfate process 2min after, outwell liquid, wipe dye vat surrounding liquid with filter paper;
Step 6, step 5 gained histotomy is put into the silvering solution of newly joining, after 37 DEG C of lucifuges dyeing 10min, distilled water is anxious washes 2 times, wipes silvering solution around dye vat with filter paper;
Step 7, step 6 gained histotomy is put into the reducing solution of newly joining, after 37 DEG C of reduction 1min, distilled water is anxious washes 2 times, histotomy is put into another dry staining rack, dries excessive moisture;
Step 8, by concentration be 75% alcohol 10s, the concentration alcohol 30s that is 85%, the concentration alcohol 2min that is 95%, concentration be 100% dimethylbenzene I 5min, concentration be that the gradient of the dimethylbenzene II 5min of 100% is dewatered to histotomy, put into baking oven 10min, acceleration of alcohol volatilizees;
Step 9, the histotomy after drying is placed in dimethylbenzene I and each 8min of dimethylbenzene II successively, after carrying out transparent processing, adopts neutral resins mounting.
Wherein, described silvering solution is by the AgNO of 3ml 0.01g/ml
3, 10ml PH is the acetate buffer solution of 5.6, the distilled water of 87ml mixes.
Wherein, described reducing solution is by p-dihydroxy-benzene 1g, anhydrous sodium sulfite 2.5g, and adding distil water is settled to 100ml gained.
The invention has the beneficial effects as follows:
Technique is simple, and easy to operate, Color is good, and colouring required time is short.
Embodiment
In order to make objects and advantages of the present invention clearly understand, below in conjunction with embodiment, the present invention is further elaborated.Should be appreciated that specific embodiment described herein only in order to explain the present invention, be not intended to limit the present invention.
Embodiments provide a kind of rapid dyeing method of endocrine cell, it is characterized in that, comprise the steps:
Step 1, histotomy is put into staining rack be placed in 100% dimethylbenzene I, each 8min of 100% dimethylbenzene II successively;
Step 2, by the histotomy of step 1 gained by concentration be 100% dimethylbenzene I 5min, concentration to be 100% dimethylbenzene II 5min concentration be 95% alcohol 5min, concentration be 85% alcohol 5min, concentration be after the gradient of the alcohol 5min of 75% carries out rehydration, be placed in distilled water and clean 5min, then wash once with distilled water is anxious;
Step 3, the histotomy of step 2 gained is placed in the dye vat of the silvering solution being equipped with 60 DEG C, 60 DEG C of constant temperature lucifuges dyeing 3h, after dye vat cooling, distilled water is anxious washes 2 times, wipes silvering solution around dye vat with filter paper;
Step 4, the histotomy of step 3 gained put into the reducing solution of 45 DEG C, after 45 DEG C of constant temperature reduction 1h, distilled water is anxious washes 2 times;
Step 5, by reduction after histotomy put into concentration be 5% sodium thiosulfate process 2min after, outwell liquid, wipe dye vat surrounding liquid with filter paper;
Step 6, step 5 gained histotomy is put into the silvering solution of newly joining, after 37 DEG C of lucifuges dyeing 10min, distilled water is anxious washes 2 times, wipes silvering solution around dye vat with filter paper;
Step 7, step 6 gained histotomy is put into the reducing solution of newly joining, after 37 DEG C of reduction 1min, distilled water is anxious washes 2 times, histotomy is put into another dry staining rack, dries excessive moisture;
Step 8, by concentration be 75% alcohol 10s, the concentration alcohol 30s that is 85%, the concentration alcohol 2min that is 95%, concentration be 100% dimethylbenzene I 5min, concentration be that the gradient of the dimethylbenzene II 5min of 100% is dewatered to histotomy, put into baking oven 10min, acceleration of alcohol volatilizees;
Step 9, by dry after histotomy be placed in dimethylbenzene I and each 8min of dimethylbenzene II successively, after carrying out transparent processing, visual inspection histotomy is in yellow, and under light microscopic, tissues observed background colour is faint yellow, positive cell is brown color, adopts neutral resins mounting.
Described silvering solution is by the AgNO of 3ml 0.01g/ml
3, 10ml PH is the acetate buffer solution of 5.6, the distilled water of 87ml mixes.Wherein, described reducing solution is by p-dihydroxy-benzene 1g, anhydrous sodium sulfite 2.5g, and adding distil water is settled to 100ml gained.
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the premise without departing from the principles of the invention; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (3)
1. a rapid dyeing method for endocrine cell, is characterized in that, comprise the steps:
Step 1, histotomy is put into staining rack be placed in 100% dimethylbenzene I, each 8min of 100% dimethylbenzene II successively;
Step 2, by the histotomy of step 1 gained by concentration be 100% dimethylbenzene I 5min, concentration to be 100% dimethylbenzene II 5min concentration be 95% alcohol 5min, concentration be 85% alcohol 5min, concentration be after the gradient of the alcohol 5min of 75% carries out rehydration, be placed in distilled water and clean 5min, then wash once with distilled water is anxious;
Step 3, the histotomy of step 2 gained is placed in the dye vat of the silvering solution being equipped with 60 DEG C, 60 DEG C of constant temperature lucifuges dyeing 3h, after dye vat cooling, distilled water is anxious washes 2 times, wipes silvering solution around dye vat with filter paper;
Step 4, the histotomy of step 3 gained put into the reducing solution of 45 DEG C, after 45 DEG C of constant temperature reduction 1h, distilled water is anxious washes 2 times;
Step 5, by reduction after histotomy put into concentration be 5% sodium thiosulfate process 2min after, outwell liquid, wipe dye vat surrounding liquid with filter paper;
Step 6, step 5 gained histotomy is put into the silvering solution of newly joining, after 37 DEG C of lucifuges dyeing 10min, distilled water is anxious washes 2 times, wipes silvering solution around dye vat with filter paper;
Step 7, step 6 gained histotomy is put into the reducing solution of newly joining, after 37 DEG C of reduction 1min, distilled water is anxious washes 2 times, histotomy is put into another dry staining rack, dries excessive moisture;
Step 8, by concentration be 75% alcohol 10s, the concentration alcohol 30s that is 85%, the concentration alcohol 2min that is 95%, concentration be 100% dimethylbenzene I 5min, concentration be that the gradient of the dimethylbenzene II 5min of 100% is dewatered to histotomy, put into baking oven 10min, acceleration of alcohol volatilizees;
Step 9, the histotomy after drying is placed in dimethylbenzene I and each 8min of dimethylbenzene II successively, after carrying out transparent processing, adopts neutral resins mounting.
2. the rapid dyeing method of a kind of endocrine cell according to claim 1, is characterized in that, described silvering solution is by the AgNO of 3ml0.01g/ml
3, 10ml PH is the acetate buffer solution of 5.6, the distilled water of 87ml mixes.
3. the rapid dyeing method of a kind of endocrine cell according to claim 1, is characterized in that, described reducing solution is by p-dihydroxy-benzene 1g, anhydrous sodium sulfite 2.5g, and adding distil water is settled to 100ml gained.
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| CN201510474195.8A CN105021440A (en) | 2015-07-31 | 2015-07-31 | Endocrine cell rapid dyeing method |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1837772A (en) * | 2006-04-10 | 2006-09-27 | 暨南大学 | Quick Feulgen dyeing method |
| US20100047860A1 (en) * | 2007-03-29 | 2010-02-25 | National University Corporation University of Toya ma | Device for storing specimen slice and instrument for microscopic observation provided with the same |
| CN102062709B (en) * | 2010-12-13 | 2012-08-29 | 苏州大学 | Method for quickly slicing fish tissues |
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2015
- 2015-07-31 CN CN201510474195.8A patent/CN105021440A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1837772A (en) * | 2006-04-10 | 2006-09-27 | 暨南大学 | Quick Feulgen dyeing method |
| US20100047860A1 (en) * | 2007-03-29 | 2010-02-25 | National University Corporation University of Toya ma | Device for storing specimen slice and instrument for microscopic observation provided with the same |
| CN102062709B (en) * | 2010-12-13 | 2012-08-29 | 苏州大学 | Method for quickly slicing fish tissues |
Non-Patent Citations (3)
| Title |
|---|
| 张哲: "《实用病理组织染色技术》", 31 March 1988 * |
| 滕利荣 等: "《生物学基础实验教程》", 30 June 2008 * |
| 胡继鹰: "《医学生物学实验与习题》", 30 November 1996 * |
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Application publication date: 20151104 |