CN105131647B - A kind of dye composite, application and its application method for biological HE dyeing - Google Patents
A kind of dye composite, application and its application method for biological HE dyeing Download PDFInfo
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Abstract
The present invention discloses a kind of medical biotechnology Senile Mouse field, and in particular to a kind of dye composite, application and its application method for biological HE dyeing.The dye composite is made up of A liquid and the mixing of B liquid；A liquid includes dyeing active ingredient a, dyeing auxiliary element and water；Wherein dyeing active ingredient a is made up of haematine, alum, oxidant, and dyeing auxiliary element is made up of some components in citric acid, glycerine；B liquid is made up of dyeing active ingredient b and water, wherein dyeing active ingredient b is Yihong and 80 85% alcohol；The mixed proportion of the A liquid and B liquid is volume ratio 3:(18‑25).It can effectively reduce the staining procedure of existing HE staining techniques to dye composite of the present invention for biological HE dyeing, simplify dyeing course, be easier to grasp.
Description
Technical field
The present invention relates to a kind of medical biotechnology Senile Mouse field, and in particular to a kind of dyestuff for biological HE dyeing
Composition, application and its application method.
Background technology
Medical biotechnology Senile Mouse field usually requires to dye the morphosis such as the organelle in biological tissue,
In favor of reaching visual observation, limit and it is checked.In order to reach more favourable visual observation effect, to biology
The staining technique of tissue is constantly developing, and generally use HE staining techniques, H refers to haematine (Hemaltoxylm), E refers to Yihong
(eosin), its chemical formula difference is as follows：
Haematine is to be used for one of natural dye of biology earliest, is dye the most frequently used in biological morphology laboratory
Material, it extracts and obtained from the pith wood of bush tree, in fawn-coloured powdered, be soluble in alcohol, also dissolve in
Hot water.From the chemical constitution in Yihong, it can be seen that its acidic colorant group is hydroxyl (- OH) and carboxyl (- COOH), its energy
Generation sodium salt (- COONa) is acted on NaOH.And sodium salt is after ionization, its coloured ion is the process of anion, can be made thin
Composition in kytoplasm and extracellular matrix dyes red.
HE staining techniques are generally divided into two kinds, when traditional classical paraffin section HE colouring methods, second, existing conventional
Common monoblock hematoxylin eosin stain method.Because traditional paraffin tissue sections dyeing is all to take the side of monolithic piece dye
Method, it is a kind of traditional and the most frequently used paraffin section technology.Its base program：(i) draw materials and fixed；(ii) it is dehydrated and wraps
Bury；(iii) section, dyeing and sealing.This method in Medical Morphology field extensively using and be known.But
This facture has the drawbacks of certain and limitation, such as when the continuous Teaching Sections of high-volume are done in making；Meanwhile it can also consume
Take the substantial amounts of time and efforts of producer.Commonly the operation sequence of monoblock hematoxylin eosin stain method is usually：Gu materials →
Fixed → decolouring → mordant dyeing → brazilwood extract dyeing liquid dyeing → color separation → reduction → flowing water flushing → distillation washing → eosin stains liquid
Dyeing → routine dehydration → transparent → FFPE, section → paster and roasting piece → transparent → gummy sealing.But this method is same
There is also certain drawback, dyeing liquor staining procedure is more, the shortcomings of being not easy to grasp, if staining technique grasp is not in place, is easy for
Cause drawn materials institutional framework difference occur and the uneven consistent phenomenon of coloration result occur, but also process mordant dyeing →
The processes such as brazilwood extract dyeing liquid dyeing → color separation → reduction → washing → eosin stains liquid dyeing.
Whether prior paraffin method or monoblock HE colouring methods, dyeing course is all complex, and staining procedure is more,
It is not easy to grasp.
The content of the invention
The present inventor by years of researches find, by existing dopant composition h and E in dyeing course institute
Configure the dyeing liquor formed to be typically only capable to individually be dyed respectively, that is, need after a certain dyeing composition dyeing, to dyeing pair
Staining procedure as can just be carried out after necessarily handling another dyeing composition, the organelle for the biological tissue for otherwise dyeing out
Morphosis can not visual observation, i.e. nucleus and cytoplasmic Color is distinguished not obvious enough, up for further changing
Enter.
In view of this, the present invention provides a kind of dye composite, application and its application method for biological HE dyeing, its
The staining procedure of existing HE staining techniques can effectively be reduced, simplify dyeing course, be easier to grasp.
To solve above technical problem, the technical scheme is that using a kind of dye combinations for biological HE dyeing
Thing, the dye composite are made up of A liquid and the mixing of B liquid；
A liquid includes dyeing active ingredient a, dyeing auxiliary element and water, wherein dyeing active ingredient a, dyeing auxiliary into
Divide and the ratio of weight and number of water is (1.5-2.5):(4-5.5):(90-100)；Active ingredient a is wherein dyed by parts by weight
Than being respectively that 0.3-0.5 parts haematine, 1.2-1.8 parts alum, 0.02-0.08 parts are any selected from sodium iodate, potassium permanganate
Oxidant composition, dyeing auxiliary element be made up of some components in citric acid, glycerine；
B liquid is made up of dyeing active ingredient b and water, wherein dyeing active ingredient b is Yihong and 80-85% alcohol, and
Yihong, water, the ratio of weight and number of 80-85% alcohol are 1:(3-5):(95-100)；
The mixed proportion of the A liquid and B liquid is volume ratio 3:(18-25).
Preferably, the ratio of weight and number that active ingredient a, dyeing auxiliary element and water are dyed in the A liquid is 2.05:
5.02:95。
Preferably, it is respectively 0.4 part of haematine, 1.6 parts of sulfuric acid active ingredient a to be dyed in the A liquid by ratio of weight and number
Aluminium potassium, 0.05 part of any oxidant composition selected from sodium iodate, potassium permanganate.
Preferably, the oxidant is sodium iodate.
Preferably, it is 0.02 part of citric acid and 5 parts of glycerine groups auxiliary element to be dyed in the A liquid by ratio of weight and number
Into.
Preferably, Yihong, water, the ratio of weight and number of 85% alcohol are 1 in the B liquid:5:100.
Preferably, the mixed proportion of the A liquid and B liquid is volume ratio 3:20.
The application also provides second of technical scheme, i.e., when foregoing any described dye composite dyes as biological HE
The application of dyestuff, it is characterised in that：The biology refers mainly to biological tissue.
The application also provides the third technical scheme, i.e., foregoing any described dye composite dyes for biological HE
When application method, it is characterised in that：Biological tissue is fixed in FAA fixers first, after flowing water is washed, being put into right will
Ask and dyed in any described dye composites of 1-7, be then through dehydration, FFPE, section, drying sealing progressively
Can.
Preferably, the FAA fixers be by volume ratio be 1:1:40% formalin of (18-20), glacial acetic acid, 70%
Alcohol forms.
The basic explanation of the herein described dye composite for biological HE dyeing is as follows：
The present inventor show that dye composite described herein can be effectively to biological group by developing for a long time
Knit and dyed, gained Color is that nucleus takes on a red color in blueness, cytoplasm, and color is clearly demarcated, is advantageous to eucaryotic cell structure
Effectively observed.Herein described dye composite can effectively reduce the dyeing step of existing HE staining techniques simultaneously
Suddenly, simplify dyeing course, be easier to grasp.
Brief description of the drawings
Fig. 1-3 is liver,spleen,kidney histotomy effect diagram corresponding to the difference of dye liquor 5 in embodiment one.
Embodiment
In order that those skilled in the art more fully understands technical scheme, with reference to specific embodiment pair
The present invention is described in further detail.
Some dye composites for being used for biological HE dyeing related to the application will be enumerated below.
Dye liquor 1：
The dye composite of the dye liquor 1 is made up of A liquid and the mixing of B liquid；
A liquid is made up of dyeing active ingredient a, dyeing auxiliary element and water, wherein dyeing active ingredient a, dyeing auxiliary
The ratio of weight and number of composition and water is 1.5:5.5:100；It is respectively 0.3 part that active ingredient a, which is wherein dyed, by ratio of weight and number
Haematine, 1.8 parts of alums, 0.08 part of potassium permanganate composition, dyeing auxiliary element are made up of citric acid；
B liquid is made up of dyeing active ingredient b and water, wherein dyeing active ingredient b is Yihong and 80% alcohol, and she
Red, water, the ratio of weight and number of 80% alcohol are 1:3:95；
The mixed proportion of the A liquid and B liquid is volume ratio 3:18.
Dye liquor 2：
The dye composite of the dye liquor 2 is made up of A liquid and the mixing of B liquid；
A liquid is made up of dyeing active ingredient a, dyeing auxiliary element and water, wherein dyeing active ingredient a, dyeing auxiliary
The ratio of weight and number of composition and water is 1.5:5.5:100；It is respectively 0.3 part that active ingredient a, which is wherein dyed, by ratio of weight and number
Haematine, 1.8 parts of alums, 0.08 part of sodium iodate composition, dyeing auxiliary element are made up of glycerine；
B liquid is made up of dyeing active ingredient b and water, wherein dyeing active ingredient b is Yihong and 80% alcohol, and she
Red, water, the ratio of weight and number of 80% alcohol are 1:3:95；
The mixed proportion of the A liquid and B liquid is volume ratio 3:18.
Dye liquor 3：
The dye composite of the dye liquor 3 is made up of A liquid and the mixing of B liquid；
A liquid is made up of dyeing active ingredient a, dyeing auxiliary element and water, wherein dyeing active ingredient a, dyeing auxiliary
The ratio of weight and number of composition and water is 2.5:4:90；It is respectively 0.5 portion of bush that active ingredient a, which is wherein dyed, by ratio of weight and number
Essence, 1.2 parts of alums, 0.02 part of potassium permanganate composition, dyeing auxiliary element are made up of citric acid；
B liquid is made up of dyeing active ingredient b and water, wherein dyeing active ingredient b is Yihong and 85% alcohol, and she
Red, water, the ratio of weight and number of 85% alcohol are 1:5:100；
The mixed proportion of the A liquid and B liquid is volume ratio 3:25.
Dye liquor 4：
The dye composite of the dye liquor 4 is made up of A liquid and the mixing of B liquid；
A liquid is made up of dyeing active ingredient a, dyeing auxiliary element and water, wherein dyeing active ingredient a, dyeing auxiliary
The ratio of weight and number of composition and water is 2.5:4:90；It is respectively 0.5 portion of bush that active ingredient a, which is wherein dyed, by ratio of weight and number
Essence, 1.2 parts of alums, 0.02 part of sodium iodate composition, dyeing auxiliary element are made up of glycerine；
B liquid is made up of dyeing active ingredient b and water, wherein dyeing active ingredient b is Yihong and 85% alcohol, and she
Red, water, the ratio of weight and number of 85% alcohol are 1:5:100；
The mixed proportion of the A liquid and B liquid is volume ratio 3:25.
Dye liquor 5：
The dye composite of the dye liquor 5 is made up of A liquid and the mixing of B liquid；
A liquid is made up of dyeing active ingredient a, dyeing auxiliary element and water, wherein dyeing active ingredient a, dyeing auxiliary
The ratio of weight and number of composition and water is 2.05:5.02:95；It is respectively 0.4 part that active ingredient a, which is wherein dyed, by ratio of weight and number
Haematine, 1.6 parts of alums, 0.05 part of sodium iodate composition, dyeing auxiliary element is by 0.02 part of citric acid and 5 parts of glycerine groups
Into；
B liquid is made up of dyeing active ingredient b and water, wherein dyeing active ingredient b is Yihong and 85% alcohol, and she
Red, water, the ratio of weight and number of 85% alcohol are 1:5:100；
The mixed proportion of the 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 i.e. embodiment one to the liver,spleen,kidney tissue of rabbit, first
First pass through FAA fixers fix, haematoxylin ＆ eosin dilution dyeing liquor dyeing, tissue dewatering, FFPE, section, drying procedure,
So as to realize the display purpose to selected institutional framework and cell component.
Now particular technique method program is described below：
1. materials from three kinds of tissue samples of rabbit liver,spleen,kidney be used as Materials for slide making, draw materials size to organize 2~3mm of thickness,
Length and width 0.5cm × 0.5cm is preferred.
Tissue sample is put into FAA fixers (40% formalin 5ml, glacial acetic acid 5ml, 70% alcohol 90ml) 2. fixing
20~24 hours are fixed in wide-mouth bottle.
Washed 8~12 hours through deionization 3. embathing.
Dyed 4. dyeing immerses acquisition tissue in dye liquor 1-5 respectively, can determine to contaminate according to institutional framework feature
Color time, generally 5~12 hours.
5. embathe with deionized water rinsing 10~20 hours, then slightly washed with distilled water.
6. dehydration, with transparent each through 70%, 80%, 95% alcohol 1 hour, absolute alcohol I, II is dehydrated each 30 minutes, diformazan
Transparent each 30~40 minutes of benzene I, II.
7. waxdip and embedding through paraffin I, II, III each 30 minutes~1 hour, embedded with paraffin (54 DEG C of fusing point).
8. section with microtome thickness 5~6 μm (precision ± 1 μm), is placed into 40~45 DEG C of water temperature with exhibition piece
Open up piece.
9. paster carries out roasting piece 10~15 hours with being put into after drying paster in 50~55 DEG C of incubators.
10. dewaxing and sealing dewaxing dimethylbenzene I, II each 30 minutes, are entered using permanent mounting medium (such as neutral gum)
Row sealing.
Experimental result is shown：Show that haematoxylin ＆ eosin contaminates using the above-mentioned dye liquor 1-5 animal rabbit histotomies tested
Color, micro- Microscopic observation show that nucleus understands in blueness, kernel；Cytoplasm takes on a red color, and chromatin particle is high-visible.
Meanwhile applicants have discovered that, using dye liquor 5 compared to dyeing effect obtained by other dye liquors 1-4 in above-described embodiment one
Fruit is more prominent, embodies in the following areas.
Tissue section strain effect -1 corresponding to table 1, dye liquor 1-5
It can be seen in table 1 that compared to other l~4 group dyeing liquors, after the 5th group of dyeing liquor dyes, more can clearly observe
It is clearly more demarcated to the cell membrane of several tissues and the edge of nuclear membrane, caryoplasm.
In the 5th group of dyeing liquor, the section to tissue sample liver,spleen,kidney, Color is observed under the microscope, it is believed that
Its unique dyeing property is kept at the 1st group-the 5 group, the 5th group of dyeing liquor is that effect is most clear.For these dyeing knots
Fruit can further illustrate the concentration and Color of the blend proportion and dye liquor between haematine, oxidant, mordant, for
Develop these dyeing liquors and suffer from direct relation.
In addition, the common counter of the light measurement parameter quantitative analysis in image analysis system is also known as extinction for optical density (OD)
Degree, it refers to that light passes through the incident intensity Ia before solution or material and transmitted intensity Ib ratio of the light after
Logarithm, i.e.,：OD=1g (Ia/Ib).Have confirmed A=OD in theory (absorbance is equal to optical density).OD values are bigger, then light quilt
Degree of absorption is just bigger, and the color of material is deeper；Otherwise absorbance is smaller, then the absorbed degree of light is just smaller, thing
The color of matter is more shallow, can reflect the amount of contained substance just lower situation.It can react the shade journey of surveyed target
Degree.Using determination of absorbance can preferably the colored intensity of certain material and reflecting determine carefully in objective cell anyway
The content of certain material in born of the same parents；It is that have different absorbing wavelengths according to the staining reaction of intracellular different chemical composition, and
And certain chemical dosimetry relation between these absorption values and cytochemistry content being present, it is a kind of visible absorbance art；
Meanwhile add measure and the application of nucleus and the dulling luminosity ratio of cytoplasm coloring.
Graphical analysis (Image analysis, the IA) technology of selection, to liver,spleen,kidney tissue corresponding to 1~5 group of dye liquor
Absorbance is measured and quantitative analysis.By IA to representing absorbance cytoplasm, the cell of rabbit liver,spleen,kidney tissue respectively
The absorbance ratio that core and nucleus colour with cytoplasm is measured and analyzed, so as to inquire into several dyes being organized under IA
Color result.
Image acquistion system (OLYMPOSB × 51) is used first, is cut into slices under light microscopic by unified multiplication factor (10 × 20
Times), picture shooting is carried out to every section；The Image-Pro Plus figures produced again with Media Cybertics companies of the U.S.
As analysis software (Version 6.0), comprehensive morphological measurement analysis is carried out to picture, selects optical density (Optical
Density), to choosing positive stained area to be measured, data result is directly logged on in Excel tables, carries out statistical disposition.
The data obtained is finally transferred to SPSS16.0 versions statistical software from Excel tables, first carries out homogeneity of variance analysis, when variance is neat,
Carry out One-Way ANOVA inspections, and P<When 0.05, being calculated as difference has statistical significance, and statistics is usedRepresent.
Every kind of solution in 5 groups of dyeing liquors to liver,spleen,kidney dye to the result of acquisition, has carried out nucleus respectively
With matter color dulling luminosity ratio ratio, the measure of nucleus color absorbance and cytoplasm color absorbance, gained measurement result
It see the table below.
Tissue section strain effect -2 corresponding to table 2, dye liquor 1-5
By the way that the statistical analysis of liver,spleen,kidney tissue is handled and compared, this method can be further demonstrated that to rabbit
Liver,spleen,kidney tissue carries out the reliability of the result of disposable monoblock HE colouring methods, and especially the 5th group of dyeing liquor is most preferably applicable
Dyed in this method.The bright-coloured degree, readability and repeat usage of 5 groups of colorings of dye liquor are superior to dye liquor the 1st after dyeing
~4 groups.
Dyed effect analysis, it can be seen that the Color obtained through dye liquor 1~5 is similar, and can reach the application
Purpose described in scheme；The dyeing liquor of dye liquor 1 and 2 situation preferably for being used to show nucleus, and through the dyeing of dye liquor 3 and 4
Histocyte core and kytoplasm dyeing after liquid dyeing is partially deep, causes the poor contrast of nuclear staining and kytoplasm dyeing, institutional framework
The relative dye liquor 5 of observation Color for be not apparent from；The Color of dye liquor 5 is optimal, and nucleus is in bright-coloured indigo plant
Color, kernel, nuclear membrane and nuclear chromatin particle are high-visible, differentiate well, and endochylema pink, karyon, endochylema are with distinct contrast, most suitable
In the making of disposable monoblock HE colouring methods, reference can be made to table 1, consistent with the cell dyeing effect of light Microscopic observation；It can
Institutional framework is more clearly shown, and can distinguishes the Morphological Features of cell, can reach the mesh for improving experimental teaching and scientific research quality
's.
From above table as can be seen that histotomy corresponding to dye liquor 5 Color center-matter color contrast with
And nucleus color saturation, cytoplasm color saturation are superior to dye liquor 1-4 biopsy tissues Color.Fig. 1-3 is real
Apply liver,spleen,kidney histotomy effect diagram corresponding to the difference of dye liquor 5 in example one；Fig. 1 is liver slice Color, and Fig. 2 is spleen
Section statining effect, Fig. 3 are kidney segment Color.
In addition, the applicant similarly has found that the proportioning for adjusting A liquid and B liquid turns into more crucial factor, can by with
Lower experiment confirms：
Embodiment two：Each 3 pieces of rabbit liver,spleen,kidney is taken, tissue is cut into 0.8cm × 0.6cm × 0.2cm, FAA is put into and fixes
12~20 hours are fixed in liquid, 15~20 hours after flowing water rinses, it is dilute to be put in the haematoxylin ＆ eosin being configured to by following method
Release in dyeing liquor and dye 1~3 hour.
Haematoxylin ＆ eosin dilution dyeing liquid making method：Take brazilwood extract dyeing liquid (A liquid：Any A liquid proportioning in dye liquor 1-5)
10 milliliters, then take eosin stains liquid (B liquid：Any B liquid proportioning in dye liquor 1-5) 100 milliliters, A liquid and B liquid are pressed 1:10 volume ratios
Example mixing is made into haematoxylin ＆ eosin dilution dyeing liquor, you can uses.
After dyed, rinsed 20~24 hours through flowing water, water washing is distilled, step by step from 70%, 80%, 90% each liquid alcohol
By 2 hours.Tissue block is entered into 95%, 100% dehydration of alcohol each 2 hours again, after being put into dimethylbenzene transparent 16 hours, is put into soft
Wax soaks 2 times, every time 60 minutes, 2 hours totally.Hard wax soaks 2 times, and 60 minutes every time, totally 2 hours, routine paraffin wax embedded.Use slicer
Wax stone is cut into 5~6 microns thick of wax band.Wax band sub-cut is attached on slide into wax disk(-sc) one by one, through two after paster
Toluene dewaxes 2 times, each 30 minutes, neutral gum sealing.Its result of micro- Microscopic observation：Cell color is light, and Yihong contrasts
Color is also very light, and intercellular boundary is still differentiated, and cell outline still understands.Another situation be nuclear targeting not
Clearly, cytoplasmic Yihong counterstain is completely ineffective or coloring is light red, and cell boundaries are difficult to recognize, it is seen that the cellular morphology having is not
It is enough complete.
Embodiment three：Each 3 pieces of rabbit liver,spleen,kidney is taken, tissue is cut into 0.8cm × 0.6cm × 0.5cm, FAA is put into and fixes
15~20 hours are fixed in agent, tissue is taken out and is rinsed 20~24 hours through flowing water, place into the haematine prepared as follows
Dyed 1~2 day in Yihong dilution dyeing liquor.
Haematoxylin ＆ eosin dilution dyeing liquid making method：Take brazilwood extract dyeing liquid (A liquid：Any A liquid proportioning in dye liquor 1-5)
30 milliliters, then take eosin stains liquid (B liquid：Any B liquid proportioning in dye liquor 1-5) 100 milliliters, A liquid and B liquid are pressed 3:10 volume ratios
Example mixing is made into haematoxylin ＆ eosin dilution dyeing liquor, you can uses.
Rinsed 20 hours through flowing water again, distilled water is slightly washed, step by step from 70%, 80%, 90% dehydration of alcohols each 2~3 at different levels
Hour, place into 95%, 100% dehydration of alcohol each 1~2 hour, after being put into dimethylbenzene transparent 10~15 hours, be put into soft wax leaching
2 times, every time 30~40 minutes.Hard wax soaks 2 times, and 50~60 minutes every time, routine paraffin wax embedded.Wax stone is cut into 6 with slicer
The thick wax band of micron, wax band sub-cut is attached into wax disk(-sc) one by one and carried on glass, is dewaxed 2 times through dimethylbenzene after paster, each 20
~30 minutes, neutral gum sealing.Micro- Microscopic observation result：Cell color is slightly deep, and eucaryotic cell structure obscures, and cell boundaries are not
Clearly, it is seen that in intensive aterrimus nucleus, cytoplasm is in peony, and more disperses deform.
Example IV：Each 3 pieces of rabbit liver,spleen,kidney is taken, tissue is cut into 0.8cm × 0.6cm × 0.8cm, FAA is put into and fixes
In agent, after fixing 6~8 hours, rinsed 20~24 hours through flowing water, place into the haematoxylin ＆ eosin prepared by following method and dilute
Dyed 5~8 days in dyeing liquor.
Haematoxylin ＆ eosin dilutes the compound method of dyeing liquor：Take brazilwood extract dyeing liquid (A liquid：Any A liquid is matched somebody with somebody in dye liquor 1-5
Than) 40 milliliters, then take eosin stains liquid (B liquid：Any B liquid proportioning in dye liquor 1-5) 100 milliliters, A liquid and B liquid are pressed 4:10 bodies
Product ratio mixing is made into haematoxylin ＆ eosin dilution dyeing liquor, you can uses.
Rinsed 20~24 hours through flowing water again, distilled water is slightly washed, step by step 70% alcohol, 80% alcohol, 90% dehydration of alcohol
Each 2 hours, enter 95%, 100% dehydration of alcohol each 2 hours, soft wax is entered after reentering dimethylbenzene transparent 16 hours and soaks 2 times, every time 30
~40 minutes.Hard wax soaks 2 times, and 60 minutes every time, routine paraffin wax embedded.Wax stone is cut into 5~6 microns of thick waxes with slicer
Band, wax band sub-cut is attached on slide into wax disk(-sc) one by one, dewaxed 2 times through dimethylbenzene after paster, each 20~30 minutes,
Neutral gum sealing.Micro- Microscopic observation result：Nucleus, cytoplasm levelling colour cast are deep, and nucleus nuclear membrane is also difficult to differentiate, carefully
Born of the same parents' obscurity boundary is unclear, and drizzly reddish black state is presented in cytoplasm.
Above-described embodiment two-four does not meet requirement described herein for the Color of animal rabbit tissue.And this
The ratio of the described A liquid of application and B liquid uses volume ratio described herein as 3:18-25 can then obtain the application institute
The nucleus of the Color of the biological tissue stated can effectively carry out visualization resolution with cytoplasm, while can effectively contract
Subtract the staining procedure of existing HE staining techniques, simplify dyeing course, be easier to grasp.
It the above is only the preferred embodiment of the present invention, it is noted that above-mentioned preferred embodiment is not construed as pair
The limitation of the present invention, protection scope of the present invention should be defined by claim limited range.For the art
For those of ordinary skill, without departing from the spirit and scope of the present invention, some improvements and modifications can also be made, these change
Enter and retouch and also should be regarded as protection scope of the present invention.
Claims (8)
- A kind of 1. application method of dye composite for biological HE dyeing, it is characterised in that：Biological tissue is fixed first In FAA fixers, after flowing water is washed, be put into dye composite and dyed, then progressively through dehydration, FFPE, cut Piece, drying sealing；The dye composite is made up of A liquid and the mixing of B liquid：A liquid is made up of dyeing active ingredient a, dyeing auxiliary element and water, wherein dyeing active ingredient a, dyeing auxiliary element And the ratio of weight and number of water is (1.5-2.5)：(4-5.5)：(90-100)；Active ingredient a is wherein dyed by ratio of weight and number Respectively 0.3-0.5 parts haematine, 1.2-1.8 parts alum, 0.02-0.08 parts are any selected from sodium iodate, potassium permanganate Oxidant is formed, and dyeing auxiliary element is made up of some components in citric acid, glycerine；B liquid is made up of dyeing active ingredient b and water, wherein dyeing active ingredient b is Yihong and 80-85% alcohol, and Yihong, Water, the ratio of weight and number of 80-85% alcohol are 1：(3-5)：(95-100)；The mixed proportion of the A liquid and B liquid is volume ratio 3：(18-25)；The biology is biological tissue.
- 2. application method according to claim 1, it is characterised in that：Active ingredient a, dyeing auxiliary are dyed in the A liquid The ratio of weight and number of composition and water is 2.05:5.02:95.
- 3. application method according to claim 1, it is characterised in that：Active ingredient a is dyed in the A liquid by parts by weight Than being respectively 0.4 part of haematine, 1.6 parts of alums, 0.05 part of any oxidant composition selected from sodium iodate, potassium permanganate.
- 4. application method according to claim 1, it is characterised in that：The oxidant is sodium iodate.
- 5. application method according to claim 1, it is characterised in that：Auxiliary element is dyed in the A liquid by parts by weight Than being formed for 0.02 part of citric acid and 5 parts of glycerine.
- 6. application method according to claim 1, it is characterised in that：Yihong, water, the weight of 85% alcohol in the B liquid Portion rate is 1:5:100.
- 7. application method according to claim 1, it is characterised in that：The mixed proportion of the A liquid and B liquid is volume ratio 3: 20。
- 8. application method according to claim 1, it is characterised in that：The FAA fixers be by volume ratio be 1:1： 40% formalin of (18-20), glacial acetic acid, 70% alcohol composition.
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| 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 |
| 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 |
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| 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 |
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