CN105131647A - Dye composition for biological HE dyeing and its application and use method - Google Patents
Dye composition for biological HE dyeing and its application and use method Download PDFInfo
- Publication number
- CN105131647A CN105131647A CN201510456202.1A CN201510456202A CN105131647A CN 105131647 A CN105131647 A CN 105131647A CN 201510456202 A CN201510456202 A CN 201510456202A CN 105131647 A CN105131647 A CN 105131647A
- Authority
- CN
- China
- Prior art keywords
- liquid
- dyeing
- ratio
- water
- dye
- 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.)
- Granted
Links
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention relates to the field of medical biological morphology research and concretely relates to a dye composition for biological HE dyeing and its application and use method. The dye composition comprises a liquid A and a liquid B. The liquid A comprises a dyeing active ingredient a, a dyeing auxiliary ingredient and water. The dyeing active ingredient a comprises hematoxylin, aluminum potassium sulfate and an oxidizing agent. The dyeing auxiliary ingredient comprises citric acid and glycerol. The liquid B comprises a dyeing active ingredient b and water. The dyeing active ingredient b comprises eosin and alcohol with a concentration of 80-85%. A volume ratio of the liquid A to the liquid B is 3: (18-25). The dye composition for biological HE dyeing can effectively shorten the existing HE dyeing process, can simplify a dyeing process and can be grasped easily.
Description
Technical field
The present invention relates to a kind of medical biotechnology Senile Mouse field, be specifically related to a kind of dye for biological HE dye composite, application and using method thereof.
Background technology
Medical biotechnology Senile Mouse field needs to dye to morphological structures such as the organoids in biological tissue usually, is beneficial to reach visual observation, limits and checks it.In order to reach more favourable visual observation effect, to the staining technique of biological tissue in continuous development, usually adopt HE staining technique, H refers to phenodin (Hemaltoxylm), E refers to Yihong (eosin), and its chemical formula is as follows respectively:
Phenodin is the earliest for one of biological natural dyestuff, is dyestuff the most frequently used in biological morphology laboratory, and the pith wood that it derives from bush tree extracts and obtains, and in fawn-coloured Powdered, is soluble in alcohol, also dissolves in hot water.From the chemical structure in Yihong, can see that its acidic colorant group is hydroxyl (-OH) and carboxyl (-COOH), it can generate sodium salt (-COONa) with NaOH effect.And sodium salt is after ionization, its colored ion is the process of negatively charged ion, and the composition in tenuigenin and extracellular matrix can be made to dye redness.
HE staining technique is divided into two kinds usually, and one is traditional classical paraffin section HE dyeing process, and two is existing conventional common monoblock hematoxylin eosin stain methods.Because the dyeing of traditional paraffin tissue sections is all the method taking monolithic sheet to contaminate, it is a kind of tradition and the most frequently used paraffin section technology.Its base program: (i) draws materials and fix; (ii) dehydration and embedding; (iii) cut into slices, dye and sealing.The widespread use and being known by people in Medical Morphology field of this method.But this manufacture method has certain drawback and limitation, such as when Teaching Sections continuously in enormous quantities is done in making; Meanwhile, a large amount of time and efforts of producer can also be expended.The schedule of operation of common monoblock hematoxylin eosin stain method is generally: → → mordant dyeing → brazilwood extract dyeing liquid dyeing → color separation → reduction → running water → distillation washing → eosin stains liquid dyeing → conventional dehydration → transparent → paraffin embedding, section → paster and roasting sheet → transparent → gummy sealing of decolouring of drawing materials → fix.But equally also there is certain drawback in this method, staining fluid staining procedure is more, the not easily shortcoming such as grasp, if staining technique is grasped not in place, just easily cause drawn materials weave construction to occur difference and occur the uneven consistent phenomenon of coloration result, but also through processes such as mordant dyeing → brazilwood extract dyeing liquid dyeing → color separation → reduction → washing → eosin stains liquid dyeing.
No matter be prior paraffin method or monoblock HE dyeing process, dyeing course is all comparatively complicated, and staining procedure is more, not easily grasps.
Summary of the invention
The present inventor finds through years of researches, due to existing dopant composition h and E in dyeing course the staining fluid that configures usually can only dye respectively separately, namely need after a certain dyeing composition dyeing, the staining procedure of another dyeing composition just can be carried out after dyeing target is necessarily processed, otherwise the organoid morphological structure of dyeing biological tissue out cannot visual observation, namely nucleus and cytoplasmic Color are distinguished obvious not, need further improvement.
In view of this, the invention provides a kind of dye for biological HE dye composite, application and using method thereof, it effectively can reduce the staining procedure of existing HE staining technique, simplifies dyeing course, more easily grasps.
For solving above technical problem, technical scheme of the present invention adopts a kind of dye composite dyeed for biological HE, and described dye composite is mixed by A liquid and B liquid and forms;
A liquid comprises dyeing effective constituent a, dyeing ancillary component and water, and the ratio of weight and number of the effective constituent a that wherein dyes, dyeing ancillary component and water is (1.5-2.5): (4-5.5): (90-100); The effective constituent a that wherein dyes is respectively by ratio of weight and number that 0.3-0.5 part phenodin, 1.2-1.8 part potassium aluminium sulfate, 0.02-0.08 part are arbitraryly selected from sodium iodate, the oxygenant of potassium permanganate forms, and dyeing ancillary component is made up of components some in citric acid, glycerol;
B liquid is made up of dye effective constituent b and water, and the effective constituent b that wherein dyes is Yihong and 80-85% alcohol, and the ratio of weight and number of Yihong, water, 80-85% alcohol is 1:(3-5): (95-100);
The blending ratio of described A liquid and B liquid is volume ratio 3:(18-25).
Preferably, dye in described A liquid effective constituent a, dyeing ancillary component and the ratio of weight and number of water is 2.05:5.02:95.
Preferably, dyeing in described A liquid, effective constituent a is respectively 0.4 part of phenodin, 1.6 parts of potassium aluminium sulfates by ratio of weight and number, 0.05 part is arbitraryly selected from sodium iodate, the oxygenant of potassium permanganate forms.
Preferably, described oxygenant is sodium iodate.
Preferably, the ancillary component that dyes in described A liquid is that 0.02 part of citric acid and 5 parts of glycerol form by ratio of weight and number.
Preferably, in described B liquid, the ratio of weight and number of Yihong, water, 85% alcohol is 1:5:100.
Preferably, the blending ratio of described A liquid and B liquid is volume ratio 3:20.
The application also provides the second technical scheme, and namely aforementioned arbitrary described dye composite is as the application of dyestuff during biological HE dyeing, it is characterized in that: described biological know-how Yao Zhi biological tissue.
The application also provides the third technical scheme, the i.e. using method of aforementioned arbitrary described dye composite when dyeing for biological HE, it is characterized in that: first biological tissue is fixed in FAA stationary liquid, after flowing water is washed, put into the arbitrary described dye composite of claim 1-7 to dye, then progressively through dehydration, paraffin embedding, section, oven dry sealing.
Preferably, described FAA stationary liquid is 1:1:(18-20 by volume ratio) 40% formaldehyde solution, Glacial acetic acid, 70% alcohol form.
The basic explanation of the dye composite dyeed for biological HE described in the application is as follows:
The present inventor draws by developing for a long time, dye composite described in the application can effectively dye to biological tissue, and gained Color is nucleus is blue, and tenuigenin takes on a red color, color is clearly demarcated, is conducive to effectively observing cellularstructure.Described in the application, dye composite effectively can reduce the staining procedure of existing HE staining technique simultaneously, simplifies dyeing course, more easily grasps.
Accompanying drawing explanation
Fig. 1-3 is the liver,spleen,kidney tissue slice effect schematic diagram that in embodiment one, dye liquor 5 is corresponding respectively.
Embodiment
In order to make those skilled in the art understand technical scheme of the present invention better, below in conjunction with specific embodiment, the present invention is described in further detail.
Some dye composites being used for biological HE dyeing relevant to the application will be enumerated below.
Dye liquor 1:
The dye composite of this dye liquor 1 is mixed by A liquid and B liquid and forms;
A liquid is made up of the effective constituent a that dyes, dyeing ancillary component and water, and the ratio of weight and number of the effective constituent a that wherein dyes, dyeing ancillary component and water is 1.5:5.5:100; Wherein effective constituent a is respectively 0.3 part of phenodin by ratio of weight and number, 1.8 parts of potassium aluminium sulfates, 0.08 part of potassium permanganate form in dyeing, and dyeing ancillary component is made up of citric acid;
B liquid is made up of dye effective constituent b and water, and the effective constituent b that wherein dyes is Yihong and 80% alcohol, and the ratio of weight and number of Yihong, water, 80% alcohol is 1:3:95;
The blending ratio of described A liquid and B liquid is volume ratio 3:18.
Dye liquor 2:
The dye composite of this dye liquor 2 is mixed by A liquid and B liquid and forms;
A liquid is made up of the effective constituent a that dyes, dyeing ancillary component and water, and the ratio of weight and number of the effective constituent a that wherein dyes, dyeing ancillary component and water is 1.5:5.5:100; Wherein effective constituent a is respectively 0.3 part of phenodin by ratio of weight and number, 1.8 parts of potassium aluminium sulfates, 0.08 part of sodium iodate form in dyeing, and dyeing ancillary component is made up of glycerol;
B liquid is made up of dye effective constituent b and water, and the effective constituent b that wherein dyes is Yihong and 80% alcohol, and the ratio of weight and number of Yihong, water, 80% alcohol is 1:3:95;
The blending ratio of described A liquid and B liquid is volume ratio 3:18.
Dye liquor 3:
The dye composite of this dye liquor 3 is mixed by A liquid and B liquid and forms;
A liquid is made up of the effective constituent a that dyes, dyeing ancillary component and water, and the ratio of weight and number of the effective constituent a that wherein dyes, dyeing ancillary component and water is 2.5:4:90; Wherein effective constituent a is respectively 0.5 part of phenodin by ratio of weight and number, 1.2 parts of potassium aluminium sulfates, 0.02 part of potassium permanganate form in dyeing, and dyeing ancillary component is made up of citric acid;
B liquid is made up of dye effective constituent b and water, and the effective constituent b that wherein dyes is Yihong and 85% alcohol, and the ratio of weight and number of Yihong, water, 85% alcohol is 1:5:100;
The blending ratio of described A liquid and B liquid is volume ratio 3:25.
Dye liquor 4:
The dye composite of this dye liquor 4 is mixed by A liquid and B liquid and forms;
A liquid is made up of the effective constituent a that dyes, dyeing ancillary component and water, and the ratio of weight and number of the effective constituent a that wherein dyes, dyeing ancillary component and water is 2.5:4:90; Wherein effective constituent a is respectively 0.5 part of phenodin by ratio of weight and number, 1.2 parts of potassium aluminium sulfates, 0.02 part of sodium iodate form in dyeing, and dyeing ancillary component is made up of glycerol;
B liquid is made up of dye effective constituent b and water, and the effective constituent b that wherein dyes is Yihong and 85% alcohol, and the ratio of weight and number of Yihong, water, 85% alcohol is 1:5:100;
The blending ratio of described A liquid and B liquid is volume ratio 3:25.
Dye liquor 5:
The dye composite of this dye liquor 5 is mixed by A liquid and B liquid and forms;
A liquid is made up of the effective constituent a that dyes, dyeing ancillary component and water, and the ratio of weight and number of the effective constituent a that wherein dyes, dyeing ancillary component and water is 2.05:5.02:95; Wherein effective constituent a is respectively 0.4 part of phenodin by ratio of weight and number, 1.6 parts of potassium aluminium sulfates, 0.05 part of sodium iodate form in dyeing, and dyeing ancillary component is made up of 0.02 part of citric acid and 5 parts of glycerol;
B liquid is made up of dye effective constituent b and water, and the effective constituent b that wherein dyes is Yihong and 85% alcohol, and the ratio of weight and number of Yihong, water, 85% alcohol is 1:5:100;
The blending ratio of described A liquid and B liquid is volume ratio 3:20.
Embodiment one
The above-mentioned dye liquor 1-5 of the application is respectively used to carry out Coloration experiment and embodiment one to the liver,spleen,kidney tissue of rabbit, first fix through FAA stationary liquid, haematoxylin & eosin dilution staining fluid dyeing, tissue dewatering, paraffin embedding, section, baking procedure, thus realization is to the display object of selected weave construction and cellular constituent.
Now concrete technological method program is described below:
1. draw materials and select rabbit liver,spleen,kidney three kinds of tissue samples as Materials for slide making, size of drawing materials is to organize thick 2 ~ 3mm, and length and width 0.5cm × 0.5cm is good.
2. fixing the wide-necked bottle that tissue sample puts into FAA stationary liquid (40% formalin 5ml, Glacial acetic acid 5ml, 70% alcohol 90ml) is fixed 20 ~ 24 hours.
3. embathe through deionization washing 8 ~ 12 hours.
4. acquisition is organized to immerse in dye liquor 1-5 respectively and is dyeed by dyeing, can determine dyeing time, be generally 5 ~ 12 hours according to weave construction feature.
5. embathe and use deionized water rinsing 10 ~ 20 hours, more slightly wash with distilled water.
6. dehydration with transparent through 70%, 80%, 95% alcohol each 1 hour, raw spirit I, II dewaters each 30 minutes, dimethylbenzene I, II transparent each 30 ~ 40 minutes.
7. waxdip and embedding were through paraffin I, II, III each 30 minutes ~ 1 hour, embedded with paraffin (fusing point 54 DEG C).
8. section and exhibition sheet microtome thickness 5 ~ 6 μm (precision ± 1 μm), then the water temperature putting into 40 ~ 45 DEG C opens up sheet.
9. paster is put into 50 ~ 55 DEG C of incubators after paster and is carried out roasting sheet 10 ~ 15 hours with drying.
10. dewaxing and sealing dewaxing dimethylbenzene I, II each 30 minutes, uses permanent mounting medium (as neutral gum) to carry out sealing.
Experimental result shows: the animal rabbit tissue slice utilizing above-mentioned dye liquor 1-5 to test demonstrates hematoxylin eosin stain, and basis of microscopic observation draws, nucleus is in blue, and kernel is clear; Tenuigenin takes on a red color, and chromatin particle is high-visible.
Meanwhile, the applicant finds, adopt dye liquor 5 more outstanding compared to other dye liquors 1-4 gained Color in above-described embodiment one, particular embodiment in the following areas.
The tissue section strain effect-1 that table 1, dye liquor 1-5 are corresponding
As seen from Table 1, compared to other l ~ 4 group staining fluid, after the 5th group of staining fluid dyeing, more can observe the cytolemma of several tissue and the edge of nuclear membrane clearly, caryoplasm is clearly more demarcated.
In the 5th group of staining fluid, to the section of tissue sample liver,spleen,kidney, examine under a microscope Color, think the dyeing property all keeping its uniqueness the 1st group of-5 groups, the 5th group of staining fluid be effect the most clearly.Concentration and the Color of blend proportion between phenodin, oxygenant, mordant and dye liquor can be further illustrated for these coloration results, have direct relation for these staining fluids of development.
In addition, the common counter of the photoptometry parameter quantitative analysis in image analysis system is that optical density(OD) (OD) is also known as absorbancy, it refer to light passed through by the incident intensity Ia before solution or material and this light after the logarithm of transmitted intensity Ib ratio, that is: OD=1g (Ia/Ib).Confirm A=OD (absorbancy equals optical density(OD)) in theory.OD value is larger, then light is just larger by degree of absorption, and the color relation of material is darker; Otherwise absorbance is less, then the absorbed degree of light is just less, and the color relation of material is more shallow, can reflect the situation that contained amount of substance is just lower.It can react the shade degree of surveyed target.Adopt determination of absorbance can better in objective cell anyway certain material colored intensity and reflect the content of certain material in measured cell; It has different absorbing wavelength according to the staining reaction of different chemical composition in cell, and there is certain chemical dosimetry relation between these absorption values and cytochemistry content, and it is a kind of visible absorbance art; Meanwhile, mensuration and the application of nucleus and the painted dulling luminosity ratio of tenuigenin is added.
Choose image analysis (Imageanalysis, IA) technology, the absorbancy of liver,spleen,kidney tissue corresponding to 1 ~ 5 group of dye liquor is measured and quantitative analysis.By IA to representing the absorbancy cytoplasm of rabbit liver,spleen,kidney tissue, nucleus and nucleus respectively and the painted absorbance ratio of cytoplasm measures and analyzes, thus inquire into several coloration result be organized under IA.
First adopt image acquistion system (OLYMPOSB × 51), cutting into slices under light microscopic by unified magnification (10 × 20 times), carrying out picture shooting to often opening section; Again by the Image-ProPlus image analysis software (Version6.0) that MediaCybertics company of the U.S. produces, comprehensive morphological Measurement and analysis is carried out to picture, selective light density (Opticaldensity), measure choosing positive stained area, data results directly logs in in Excel table, carries out statistical treatment.Finally the data obtained is proceeded to SPSS16.0 version statistical software from Excel table, first carry out homoscedasticity analysis, when variance is neat, carry out One-WayANOVA inspection, and during P<0.05, count difference and have statistical significance, statistic data is used
represent.
Often kind of solution in 5 groups of staining fluids is carried out to liver,spleen,kidney the result obtained that dyes, and carried out nucleus and matter color dulling luminosity ratio ratio respectively, the mensuration of nucleus color absorbancy and tenuigenin color absorbancy, gained measurement result sees the following form.
The tissue section strain effect-2 that table 2, dye liquor 1-5 are corresponding
By to the statistical analysis process of liver,spleen,kidney tissue and compare, can show that the liver,spleen,kidney tissue of present method to rabbit carries out the reliability of the result of disposable monoblock HE dyeing process further, especially the 5th group of staining fluid should be applicable to the dyeing of this method.Bright-coloured degree, readability and repeat usage that after dyeing, dye liquor 5 groups is painted are all better than dye liquor 1st ~ 4 groups.
Dyed effect analysis, can find out that the Color obtained through dye liquor 1 ~ 5 is similar, all can reach the object described in the application's scheme; The staining fluid situation for showed cell core preferably of dye liquor 1 and 2, and through dye liquor 3 and 4 staining fluid dyeing after histocyte core and kytoplasm dyeing all partially dark, cause the poor contrast of nuclear staining and kytoplasm dyeing, the observation of weave construction relative to dye liquor 5 Color be not very clear; The Color of dye liquor 5 is best, nucleus is bright-coloured blueness, kernel, nuclear membrane and nuclear chromatin particle are high-visible, differentiate good, endochylema pink, karyon, endochylema are with distinct contrast, are most suitable for the making of disposable monoblock HE dyeing process, can see table 1, consistent with the cell dyeing effect of light Microscopic observation; It can more clearly display organization structure, can distinguish again the Morphological Features of cell, can reach the object improving experimental teaching and scientific research quality.
As can be seen from above table, the Color center-matter color contrast of the tissue slice of dye liquor 5 correspondence and nucleus color saturation, tenuigenin color saturation are all better than the biopsy tissues Color of dye liquor 1-4.Fig. 1-3 is the liver,spleen,kidney tissue slice effect schematic diagram that in embodiment one, dye liquor 5 is corresponding respectively; Fig. 1 is liver slice Color, and Fig. 2 is spleen section statining effect, and Fig. 3 is kidney segment Color.
In addition, the applicant finds that the proportioning adjusting A liquid and B liquid becomes comparatively crucial factor too, confirms by following experiment:
Embodiment two: get each 3 pieces of rabbit liver,spleen,kidney, tissue is cut into 0.8cm × 0.6cm × 0.2cm, put into FAA stationary liquid and fix 12 ~ 20 hours, after running water 15 ~ 20 hours, to be put in the haematoxylin & eosin dilution staining fluid be mixed with by method below dyeing 1 ~ 3 hour.
Haematoxylin & eosin dilution staining fluid compound method: get brazilwood extract dyeing liquid (A liquid: arbitrary A liquid proportioning in dye liquor 1-5) 10 milliliters, get eosin stains liquid (B liquid: arbitrary B liquid proportioning in dye liquor 1-5) 100 milliliters again, A liquid and B liquid are mixed by 1:10 volume ratio and is made into haematoxylin & eosin and dilutes staining fluid, can use.
After dyed, through running water 20 ~ 24 hours, distilled water wash, step by step from 70%, 80%, 90% each liquid alcohol through 2 hours.Again tissue block is entered each 2 hours of 95%, 100% dehydration of alcohol, put into dimethylbenzene after transparent 16 hours, put into soft wax and soak 2 times, each 60 minutes, totally 2 hours.Hard wax soaks 2 times, each 60 minutes, totally 2 hours, and routine paraffin wax embeds.With slicing machine, wax stone is cut into the wax band of 5 ~ 6 micron thickness.Wax band is divided and is cut into wax disk(-sc) one by one and is attached on slide glass, dewax 2 times through dimethylbenzene after paster, each 30 minutes, neutral gum sealing.Its result of basis of microscopic observation: cell color is light, Yihong differential staining is also very light, and intercellular boundary is still distinguishable, and cell outline still can understand.Another kind of situation is that nuclear targeting is unclear, and cytoplasmic Yihong counterstain is completely invalid or painted light red, and cell boundaries is difficult to identification, and the cellular form had as seen is sufficiently complete.
Embodiment three: get each 3 pieces of rabbit liver,spleen,kidney, tissue is cut into 0.8cm × 0.6cm × 0.5cm, put into FAA fixing agent and fix 15 ~ 20 hours, take out tissue through running water 20 ~ 24 hours, then put into the haematoxylin & eosin dilution staining fluid dyeing of preparing by following method 1 ~ 2 day.
Haematoxylin & eosin dilution staining fluid compound method: get brazilwood extract dyeing liquid (A liquid: arbitrary A liquid proportioning in dye liquor 1-5) 30 milliliters, get eosin stains liquid (B liquid: arbitrary B liquid proportioning in dye liquor 1-5) 100 milliliters again, A liquid and B liquid are mixed by 3:10 volume ratio and is made into haematoxylin & eosin and dilutes staining fluid, can use.
Again through running water 20 hours, distilled water is slightly washed, step by step from each 2 ~ 3 hours of 70%, 80%, 90% dehydration of alcohol at different levels, put into 95%, 100% dehydration of alcohol more each 1 ~ 2 hour, put into dimethylbenzene after transparent 10 ~ 15 hours, put into soft wax and soak 2 times, each 30 ~ 40 minutes.Hard wax soaks 2 times, each 50 ~ 60 minutes, and routine paraffin wax embeds.With slicing machine, wax stone is cut into the wax band of 6 micron thickness, wax band is divided and is cut into wax disk(-sc) one by one and is attached to and carries on glass, dewax 2 times through dimethylbenzene after paster, each 20 ~ 30 minutes, neutral gum sealing.Basis of microscopic observation result: cell color is slightly dark, and cellularstructure is fuzzy, and cell boundaries is unclear, visible in intensive aterrimus nucleus, tenuigenin is scarlet, and many disperses distortion.
Embodiment four: get each 3 pieces of rabbit liver,spleen,kidney, tissue is cut into 0.8cm × 0.6cm × 0.8cm, puts into FAA fixing agent, after fixing 6 ~ 8 hours, through running water 20 ~ 24 hours, then put into the haematoxylin & eosin dilution staining fluid dyeing 5 ~ 8 days by method preparation below.
The compound method of haematoxylin & eosin dilution staining fluid: get brazilwood extract dyeing liquid (A liquid: arbitrary A liquid proportioning in dye liquor 1-5) 40 milliliters, get eosin stains liquid (B liquid: arbitrary B liquid proportioning in dye liquor 1-5) 100 milliliters again, A liquid and B liquid are mixed by 4:10 volume ratio and is made into haematoxylin & eosin and dilutes staining fluid, can use.
Again through running water 20 ~ 24 hours, distilled water is slightly washed, and 70% alcohol, 80% alcohol, each 2 hours of 90% dehydration of alcohol, enter 95%, 100% dehydration of alcohol each 2 hours step by step, reenters dimethylbenzene and enters soft wax after transparent 16 hours and soak 2 times, each 30 ~ 40 minutes.Hard wax soaks 2 times, each 60 minutes, and routine paraffin wax embeds.With slicing machine, wax stone is cut into the wax band of 5 ~ 6 micron thickness, wax band is divided and is cut into wax disk(-sc) one by one and is attached on slide glass, dewax 2 times through dimethylbenzene after paster, each 20 ~ 30 minutes, neutral gum sealing.Basis of microscopic observation result: nucleus, tenuigenin levelling colour cast are dark, nucleus nuclear membrane is also difficult to differentiate, and cell boundaries is smudgy, and tenuigenin presents drizzly reddish black state.
Above-described embodiment two-four does not all meet the requirement described in the application for the Color of animal rabbit tissue.A liquid described in the application and the ratio of B liquid adopt the volume ratio described in the application then can obtain the nucleus of the Color of the biological tissue described in the application for 3:18-25 and tenuigenin effectively can carry out visual resolution, effectively can reduce the staining procedure of existing HE staining technique simultaneously, simplify dyeing course, more easily grasp.
Below be only the preferred embodiment of the present invention, it should be pointed out that above-mentioned preferred implementation should not be considered as limitation of the present invention, protection scope of the present invention should be as the criterion with claim limited range.For those skilled in the art, without departing from the spirit and scope of the present invention, can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (10)
1. for the dye composite that biological HE dyes, it is characterized in that: described dye composite is mixed by A liquid and B liquid and forms;
A liquid comprises dyeing effective constituent a, dyeing ancillary component and water, and the ratio of weight and number of the effective constituent a that wherein dyes, dyeing ancillary component and water is (1.5-2.5): (4-5.5): (90-100); The effective constituent a that wherein dyes is respectively by ratio of weight and number that 0.3-0.5 part phenodin, 1.2-1.8 part potassium aluminium sulfate, 0.02-0.08 part are arbitraryly selected from sodium iodate, the oxygenant of potassium permanganate forms, and dyeing ancillary component is made up of components some in citric acid, glycerol;
B liquid is made up of dye effective constituent b and water, and the effective constituent b that wherein dyes is Yihong and 80-85% alcohol, and the ratio of weight and number of Yihong, water, 80-85% alcohol is 1:(3-5): (95-100);
The blending ratio of described A liquid and B liquid is volume ratio 3:(18-25).
2. dye composite according to claim 1, is characterized in that: the ratio of weight and number of the effective constituent a that dyes in described A liquid, dyeing ancillary component and water is 2.05:5.02:95.
3. dye composite according to claim 1, is characterized in that: dyeing in described A liquid, effective constituent a is respectively 0.4 part of phenodin, 1.6 parts of potassium aluminium sulfates by ratio of weight and number, 0.05 part is arbitraryly selected from sodium iodate, the oxygenant of potassium permanganate forms.
4. dye composite according to claim 1, is characterized in that: described oxygenant is sodium iodate.
5. dye composite according to claim 1, is characterized in that: the ancillary component that dyes in described A liquid is that 0.02 part of citric acid and 5 parts of glycerol form by ratio of weight and number.
6. dye composite according to claim 1, is characterized in that: in described B liquid, the ratio of weight and number of Yihong, water, 85% alcohol is 1:5:100.
7. dye composite according to claim 1, is characterized in that: the blending ratio of described A liquid and B liquid is volume ratio 3:20.
8. the arbitrary described dye composite of claim 1-7 is as the application of dyestuff during biological HE dyeing, it is characterized in that: described biology is biological tissue.
9. the using method of the arbitrary described dye composite of claim 1-7 when dyeing for biological HE, it is characterized in that: first biological tissue is fixed in FAA stationary liquid, after flowing water is washed, put into the arbitrary described dye composite of claim 1-7 to dye, then progressively through dehydration, paraffin embedding, section, oven dry sealing.
10. using method according to claim 10, is characterized in that: described FAA stationary liquid is 1:1:(18-20 by volume ratio) 40% formaldehyde solution, Glacial acetic acid, 70% alcohol form.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510456202.1A CN105131647B (en) | 2015-07-29 | 2015-07-29 | A kind of dye composite, application and its application method for biological HE dyeing |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510456202.1A CN105131647B (en) | 2015-07-29 | 2015-07-29 | A kind of dye composite, application and its application method for biological HE dyeing |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105131647A true CN105131647A (en) | 2015-12-09 |
| CN105131647B CN105131647B (en) | 2017-12-05 |
Family
ID=54717239
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510456202.1A Expired - Fee Related CN105131647B (en) | 2015-07-29 | 2015-07-29 | A kind of dye composite, application and its application method for biological HE dyeing |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105131647B (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107490511A (en) * | 2017-07-25 | 2017-12-19 | 四川金域医学检验中心有限公司 | A kind of haematoxylin dyeing liquid and HE colouring methods |
| CN108007754A (en) * | 2016-10-31 | 2018-05-08 | 陈石磊 | A kind of one step decoration method of cast-off cells, dye combinations used and kit |
| CN110079116A (en) * | 2019-04-18 | 2019-08-02 | 中国人民解放军第八一医院 | The reactive monoazo dyestuffs TB-EMB of caryoplasm simultaneously dyeing |
| CN110887719A (en) * | 2019-12-20 | 2020-03-17 | 云南中医药大学 | Method for processing tissue sample of histopathology |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103697732A (en) * | 2013-12-25 | 2014-04-02 | 南宁广发重工集团有限公司 | Cooling barrel for multi-barrel cooler |
-
2015
- 2015-07-29 CN CN201510456202.1A patent/CN105131647B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103697732A (en) * | 2013-12-25 | 2014-04-02 | 南宁广发重工集团有限公司 | Cooling barrel for multi-barrel cooler |
Non-Patent Citations (3)
| Title |
|---|
| 旺庆 等: ""动物组织块的苏木精-曙红整块染色法"", 《解剖学杂志-中国解剖学会2002年年会文摘汇编》 * |
| 柴建权: ""苏木素伊红混合液一分钟快速染色法"", 《陕西医药杂志》 * |
| 涂琦 等: ""苏木素伊红染色一步法的研究及应用"", 《江西医药》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108007754A (en) * | 2016-10-31 | 2018-05-08 | 陈石磊 | A kind of one step decoration method of cast-off cells, dye combinations used and kit |
| CN107490511A (en) * | 2017-07-25 | 2017-12-19 | 四川金域医学检验中心有限公司 | A kind of haematoxylin dyeing liquid and HE colouring methods |
| CN107490511B (en) * | 2017-07-25 | 2019-05-07 | 四川金域医学检验中心有限公司 | A kind of haematoxylin dyeing liquid and HE colouring method |
| CN110079116A (en) * | 2019-04-18 | 2019-08-02 | 中国人民解放军第八一医院 | The reactive monoazo dyestuffs TB-EMB of caryoplasm simultaneously dyeing |
| CN110887719A (en) * | 2019-12-20 | 2020-03-17 | 云南中医药大学 | Method for processing tissue sample of histopathology |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105131647B (en) | 2017-12-05 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107490511B (en) | A kind of haematoxylin dyeing liquid and HE colouring method | |
| CN106644656A (en) | Hematoxylin-eosin one-step dyeing method | |
| CN105131647A (en) | Dye composition for biological HE dyeing and its application and use method | |
| CN103415762A (en) | Hematoxylin staining method | |
| CN111811909A (en) | Preparation and application of tissue autofluorescence quencher | |
| CN105842037B (en) | Colouring method that is a kind of while showing mast cell and acidophic cell | |
| CN109612807A (en) | A kind of urinary formed element dyeing liquor | |
| RU2536502C2 (en) | Cytological and histological fixing composition and staining method | |
| Dapson et al. | Hematoxylin shortages: their causes and duration, and other dyes that can replace hemalum in routine hematoxylin and eosin staining | |
| CN103884560A (en) | Preparation method and application of Chinese giant salamander phalanx section | |
| Flowers et al. | Colouring of Pacific barkcloths: identification of the brown, red and yellow colourants used in the decoration of historic Pacific barkcloths | |
| CN108680418A (en) | A kind of rapid fluorescence colouring method of crop in cruciferae pollen | |
| Alshamar et al. | Use of roselle extracted from Hibiscus sabdariffa for histological staining: a critical review and rational stain formulation | |
| Itodo et al. | Phytochemical properties and staining ability of red onion (Allium cepa) extract on histological sections | |
| CN109187127A (en) | Observe the preparation method of the slice dyed blended liquid and slice of xylem parenchyma | |
| Sánchez-Hernández et al. | Formalin fixation and paraffin embedding interfere with the preservation of optical metabolic assessments based on endogenous NAD (P) H and FAD two-photon excited fluorescence | |
| CN106323723B (en) | Double indigo plant decoration methods | |
| CN101804082B (en) | Method for identifying cordyceps sinensis powder through microscopic dyeing | |
| CN104390834A (en) | Sarranine and methyl violet mixed staining method for resin slices and staining solution thereof | |
| JPS6047960A (en) | Staining method and staining test solution of cell in cytology | |
| CN102589956B (en) | Method for identifying cordyceps sinensis through microscopic staining | |
| Stefanović et al. | Romanowsky-Giemsa as a counterstain for immunohistochemistry: optimizing a traditional reagent | |
| Benard | Iron-roselle: A progressive nuclear stain substitute for hematoxylin | |
| RU2593343C1 (en) | Method for staining the histological cuts in diagnosing trichinosis | |
| CN114279795A (en) | Rapid detection system, detection method and application of tissue sample |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20171205 Termination date: 20210729 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |