CN104849126A - Rapid dyeing method for mucous cells - Google Patents

Abstract

The invention discloses a rapid dyeing method for mucous cells. The rapid dyeing method comprises the following steps: putting tissue sections on a staining rack, and then sequentially putting the staining rack in 100% xylene I and 100% xylene II respectively for 8 minutes; carrying out rehydration on the tissue sections according to a gradient of 5 minutes in 100% xylene I, 5 minutes in 100% xylene II, 5 minutes in 95% alcohol, 5 minutes in 85% alcohol and 5 minutes in 75% alcohol, and then rinsing; taking two pieces of sections, washing the tissue sections for three times by using distilled water dropwise, and spin-drying; taking one to two drops of 1% Alcian blue dyeing liquid with a dropper and dyeing the tissue sections, then rinsing, and washing tissues for three times by using distilled water dropwise; taking one to two drops of 1% periodic acid solution with a dropper and dyeing dropwise, then rinsing, and washing the tissues for three times by using distilled water dropwise; then taking one to two drops of 0.5% Schiff reagent with a dropper and dyeing the tissue sections dropwise, throwing away excessive Schiff reagent on the sections, putting the sections on the staining rack and putting the staining rack in a beaker, dropping water to return red, dehydrating and drying, then carrying out transparent processing, and sealing with neutral resin.

Description

A kind of rapid dyeing method of mucilage cell

Technical field

The present invention relates to cell dyeing field, be specifically related to a kind of rapid dyeing method of mucilage 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 mucilage cell.

For achieving the above object, the technical scheme that the present invention takes is:

A rapid dyeing method for mucilage 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 histotomy 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 tap water running water 5min;

Step 3, get 2 section distillation water droplets and wash histotomy 3 times, dry;

Step 4, to get two histotomies that concentration is 1%, PH is 2.5 1% A Li Xinlan dye liquor drips dye step 3 gained with dropper, dyeing 5min.

Step 5, the histotomy of step 4 gained low-speed water flow to be rinsed after 2min, wash tissue 3 times with distillation water droplet, with ear washing bulb, moisture around tissue is dried up;

Step 6, to get with dropper concentration be 0.5% 1% periodic acid solution drip dye step 5 gained histotomy, oxidation 2min;

Step 7, the histotomy of step 6 gained low-speed water flow to be rinsed after 1min, wash tissue 3 times with distillation water droplet, with ear washing bulb, moisture around tissue is dried up;

Step 8, get 0.5% snow husband reagent with dropper and drip dye histotomy, dyeing 5min;

Step 9, get rid of the upper unnecessary snow husband reagent of section, staining rack is put in section and is placed in beaker, drip and return red 9-11min, by chromatic effect in microscopic examination, if poor effect, proceed to return red, until paint; (note: first add a small amount of tap water in beaker and flood tissue, drips and to avoid liquid to flow out beaker when returning red as far as possible, tissue is in and returns in red water environment, return red effect better, return red after can not rinse, Direct Dehydration).

Step 10, 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 5min, acceleration of alcohol volatilizees;

Step 11, 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.

Preferably, described A Li Xinlan dye liquor, by A Li Xinlan 1g, adds 3% aqueous acetic acid and is settled to 100ml gained.

The invention has the beneficial effects as follows:

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 mucilage cell, 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 histotomy 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 tap water running water 5min;

Step 3, get 2 section distillation water droplets and wash histotomy 3 times, dry;

Step 4, to get two histotomies that concentration is 1%, PH is 2.5 1% A Li Xinlan dye liquor drips dye step 3 gained with dropper, dyeing 5min.

Step 5, the histotomy of step 4 gained low-speed water flow to be rinsed after 2min, wash tissue 3 times with distillation water droplet, with ear washing bulb, moisture around tissue is dried up;

Step 6, to get with dropper concentration be 0.5% 1% periodic acid solution drip dye step 5 gained histotomy, oxidation 2min;

Step 7, the histotomy of step 6 gained low-speed water flow to be rinsed after 1min, wash tissue 3 times with distillation water droplet, with ear washing bulb, moisture around tissue is dried up;

Step 8, get 0.5% snow husband reagent with dropper and drip dye histotomy, dyeing 5min;

Step 9, get rid of the upper unnecessary snow husband reagent of section, staining rack is put in section and is placed in beaker, drip and return red 9-11min, by chromatic effect in microscopic examination, if poor effect, proceed to return red, until paint; (note: first add a small amount of tap water in beaker and flood tissue, drips and to avoid liquid to flow out beaker when returning red as far as possible, tissue is in and returns in red water environment, return red effect better, return red after can not rinse, Direct Dehydration).

Step 10, 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 5min, acceleration of alcohol volatilizees;

Step 11, 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.

Described A Li Xinlan dye liquor, by A Li Xinlan 1g, adds 3% aqueous acetic acid and is settled to 100ml gained.

After testing, AB-PAS dye goblet cell result is divided into 4 types:

I type takes on a red color, and AB is negative, and PAS is positive, containing neutral mucopolysacchauide.

II type is in blue, and AB is positive, and PAS is negative, containing acid mucopolysaccharide.

Type III is aubergine, and AB and PAS is the positive, the main neutral mucopolysacchauide containing the PAS positive, the acid mucopolysaccharide simultaneously containing a small amount of AB positive.

IV type is bluish violet, and AB and PAS is the positive, the main acid mucopolysaccharide containing the AB positive, the neutral mucopolysacchauide simultaneously containing a small amount of PAS positive.

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 (2)

1. a rapid dyeing method for mucilage 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 histotomy 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 tap water running water 5min;

Step 3, get 2 section distillation water droplets and wash histotomy 3 times, dry;

Step 4, to get two histotomies that concentration is 1%, PH is 2.5 1% A Li Xinlan dye liquor drips dye step 3 gained with dropper, dyeing 5min.

Step 5, the histotomy of step 4 gained low-speed water flow to be rinsed after 2min, wash tissue 3 times with distillation water droplet, with ear washing bulb, moisture around tissue is dried up;

Step 6, to get with dropper concentration be 0.5% 1% periodic acid solution drip dye step 5 gained histotomy, oxidation 2min;

Step 7, the histotomy of step 6 gained low-speed water flow to be rinsed after 1min, wash tissue 3 times with distillation water droplet, with ear washing bulb, moisture around tissue is dried up;

Step 8, get 0.5% snow husband reagent with dropper and drip dye histotomy, dyeing 5min;

Step 9, get rid of the upper unnecessary snow husband reagent of section, staining rack is put in section and is placed in beaker, drip and return red 9-11min, by chromatic effect in microscopic examination, if poor effect, proceed to return red, until paint;

Step 10, 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 5min, acceleration of alcohol volatilizees;

Step 11, 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 mucilage cell according to claim 1, is characterized in that, described A Li Xinlan dye liquor, by A Li Xinlan 1g, adds 3% aqueous acetic acid and is settled to 100ml gained.