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CN-122016442-A - Economical collagen fiber dyeing kit and preparation method thereof

CN122016442ACN 122016442 ACN122016442 ACN 122016442ACN-122016442-A

Abstract

The invention relates to the technical field of biology, in particular to an economic collagen fiber dyeing kit and a preparation method thereof, wherein the kit comprises the following reagents: modified hematoxylin, cole's hematoxylin solution, sirius scarlet staining solution, modified eosin staining solution, modified multiple red solution, differentiation concentrate, acidic differentiation solution, bluing solution and Scott bluing solution. The modified iron hematoxylin a solution can enhance the dissolubility of dye, delay oxidation browning and prevent precipitation, so that nuclear dyeing is clearer and more uniform, the modified iron hematoxylin b solution can stabilize iron ion release, reduce excessive corrosiveness of hydrochloric acid and enhance dyeing contrast, and the modified iron hematoxylin b solution are mixed to ensure that cell nuclear dyeing has excellent selectivity, vivid contrast and good reproducibility, the modified eosin dye solution can maintain stable pH of dye solution and enhance permeability and affinity to cytoplasm, and the modified eosin solution can quickly form uniform and stable working solution to realize dyeing effects of strong specificity and vivid contrast on collagen fibers.

Inventors

  • ZHAN SHUAISHUAI
  • CHEN JINGLI
  • WANG XIAOYONG

Assignees

  • 青岛恒星科技学院

Dates

Publication Date
20260512
Application Date
20260312

Claims (10)

  1. 1. A preparation method of an economic collagen fiber dyeing reagent is characterized by comprising the following reagents of modified hematoxylin, cole's hematoxylin liquid, sirius red dyeing liquid, modified eosin dyeing liquid, modified multiple red liquid, differentiation concentrate, acidic differentiation liquid, bluing liquid and Scott blue-promoting liquid; The modified hematoxylin is prepared by mixing a modified hematoxylin a solution and a modified hematoxylin b solution in equal volume ratio of 1:1; the modified multiple red liquid is prepared by mixing a modified multiple red dye liquid a and a modified multiple red diluent liquid b according to a volume ratio of 1:9.
  2. 2. The preparation method of the economic collagen fiber dyeing reagent according to claim 1, wherein the preparation method of the modified hematoxylin a solution is characterized in that hematoxylin is added into absolute ethyl alcohol at 25-45 ℃ in a water bath, stirring is continued until the hematoxylin is completely dissolved, then glycerol, ascorbic acid and polyethylene glycol 400 are sequentially added, stirring is continued for 5-10min until the mixture is uniformly mixed after each component is added, after all components are added, the mixed solution is transferred into a light-proof container, stirring is continued at room temperature, and then sealing and light-proof standing and curing are carried out for 4-6 weeks at 4 ℃ to obtain the modified hematoxylin a solution.
  3. 3. The preparation method of the economic collagen fiber dyeing reagent according to claim 2, wherein the mass ratio of hematoxylin to absolute ethyl alcohol is 1 (75-85), the mass ratio of hematoxylin to glycerol is 1 (0.6-1), the mass ratio of hematoxylin to ascorbic acid is 1 (0.04-0.06), and the mass ratio of hematoxylin to polyethylene glycol 400 is 1 (2-3).
  4. 4. The preparation method of the economic collagen fiber dyeing reagent according to claim 1, wherein the preparation method of the modified hematoxylin b solution is characterized in that deionized water is added into 30% ferric trichloride solution by mass fraction and stirred uniformly, then concentrated hydrochloric acid is added dropwise under continuous stirring, the pH value is controlled to be 1.5-2.5, sodium citrate and aluminum potassium sulfate powder are added sequentially after the dropwise addition is completed, and the modified hematoxylin b solution is prepared after continuous stirring until the solid is completely dissolved and the solution is clear.
  5. 5. The method for preparing an economic collagen fiber dyeing reagent according to claim 4, wherein the mass ratio of the ferric trichloride solution to the deionized water is 1 (0.6-0.7), the mass ratio of the ferric trichloride solution to the concentrated hydrochloric acid is 1 (0.2-0.25), the mass ratio of the ferric trichloride solution to the sodium citrate is 1 (0.05-0.07), and the mass ratio of the ferric trichloride solution to the aluminum potassium sulfate powder is 1 (0.02-0.04).
  6. 6. The preparation method of the economic type collagen fiber dyeing reagent according to claim 1, wherein the preparation method of the modified eosin dye solution is characterized in that 95-100 parts of acetic acid-sodium acetate buffer solution with the pH value of 4.8 and 1 part of water-soluble eosin Y powder are mixed, and continuously stirred to form uniform red clear solution, 0.08-0.12 part of polysorbate-20 and 0.4-0.6 part of benzyl alcohol are sequentially added into the clear solution, and continuously stirred until the added components are fully mixed, and finally the volume is fixed and homogenized to prepare the modified eosin dye solution.
  7. 7. The preparation method of the economic type collagen fiber dyeing reagent according to claim 1 is characterized in that 95-105 parts by mass of 1% acetic acid aqueous solution and 1 part of acid fuchsin powder are mixed, continuously stirred until the powder is dissolved, then 0.02-0.03 part of sodium benzoate and 0.08-0.12 part of polyethylene glycol 6000 are sequentially added, and continuously stirred until the added components are uniformly dispersed and dissolved, so that the modified multiple red dye liquor a is prepared.
  8. 8. The preparation method of the economic type collagen fiber dyeing reagent according to claim 1 is characterized in that the preparation method of the modified reddish-red diluent b comprises the steps of mixing 1 part of saturated picric acid aqueous solution with 0.10-0.15 part of ethanol solution with the mass fraction of 70%, stirring until the saturated picric acid aqueous solution and the ethanol solution are fully mixed, then adding 0.010-0.015 part of glacial acetic acid solution under the stirring condition, controlling the pH value to be stable to 2.0-2.5, adding 0.007-0.009 part of saturated sodium chloride aqueous solution, stirring at a rising speed until all components are completely dissolved and the solution is clear, and after the mixing is completed, reducing the rotating speed, continuously stirring to complete homogenization, so as to obtain the modified reddish-red diluent b.
  9. 9. The method for preparing an economical dyeing reagent for collagen fibers according to claim 8, wherein the saturated aqueous solution of picric acid is prepared by adding 0.06-0.08 parts of picric acid crystals to 1 part of distilled water, continuously stirring until the picric acid crystals are completely dissolved, adding 0.03-0.05 parts of the distilled water as a standard, stirring until the picric acid crystals are completely dissolved, and the solution is in a clear state, repeating the process, indicating that the solution is saturated when solid residues are observed at the bottom of the container and stirring is not dissolved, stopping stirring, standing, and filtering to obtain the supernatant.
  10. 10. An economic collagen fiber dyeing kit is prepared by a preparation method of the economic collagen fiber dyeing reagent according to any one of claims 1-9, and is characterized in that 11 drip bottles are filled in the kit, different reagents are filled in the drip bottles, and the reagents in the 11 drip bottles are respectively modified iron hematoxylin a solution, modified iron hematoxylin b solution, cole's hematoxylin solution, sirius scarlet dyeing solution, modified eosin dyeing solution, modified carmine dyeing solution a, modified carmine diluting solution b, differentiation concentrate, acidic differentiation solution, blueing solution and Scott blue-promoting solution.

Description

Economical collagen fiber dyeing kit and preparation method thereof Technical Field The invention relates to the technical field of biology, in particular to an economic collagen fiber dyeing kit and a preparation method thereof. Background The reagent can effectively distinguish collagen fibers from other components such as myofibers and the like, and is mainly used for evaluating the degree of tissue fibrosis, assisting in tumor differential diagnosis and researching the tissue repair process. In the prior art, the conventional iron hematoxylin a solution is easy to oxidize and brown, poor in stability, and excessive corrosion and uneven mordant dyeing of tissues are easy to occur due to over-strong acidity of b solution, and precipitation or inconsistent dyeing often occurs when the b solution and the b solution are mixed, the pH of the common eosin dye solution is unstable and easy to precipitate, uneven cytoplasmic dyeing and residual background are caused, HE dyeing definition and reproducibility are affected, and the common multiple-red dye solution and picric acid diluent are poor in mixing compatibility, and problems of insufficient dissolution, uneven dyeing and the like are easy to occur, so that the dyeing contrast of collagen fibers is unclear and the fluctuation of results is large. Based on the above, the invention provides an economic collagen fiber dyeing kit and a preparation method thereof. Disclosure of Invention The invention aims to provide an economic collagen fiber dyeing kit and a preparation method thereof, wherein the prepared dyeing reagent can be stored for a long time, so that the stability is better, the value of a variation coefficient CV% is lower, the dyeing result is uniform, the color contrast index DeltaOD is higher, the object and the background are obviously distinguished after dyeing, and the image definition is high. The invention provides an economic collagen fiber dyeing kit and a preparation method thereof, wherein the economic collagen fiber dyeing kit comprises modified hematoxylin, cole's hematoxylin solution, sirius red dyeing solution, modified eosin dyeing solution, modified multiple red solution, differentiation concentrate, acidic differentiation solution, bluing solution and Scott blue-promoting solution; The modified hematoxylin is prepared by mixing a modified hematoxylin a solution and a modified hematoxylin b solution in equal volume ratio of 1:1; the modified multiple red liquid is prepared by mixing a modified multiple red dye liquid a and a modified multiple red diluent liquid b according to a volume ratio of 1:9. The preparation method of the modified hematoxylin a solution comprises the steps of adding hematoxylin in absolute ethyl alcohol at 25-45 ℃ in a water bath, continuously stirring until the hematoxylin is completely dissolved, sequentially adding glycerol, ascorbic acid and polyethylene glycol 400, continuously stirring for 5-10min until the components are uniformly mixed, transferring the mixed solution into a light-proof container after all the components are added, continuously stirring at room temperature, sealing, and standing and curing for 4-6 weeks at 4 ℃ in a light-proof mode to obtain the modified hematoxylin a solution. Preferably, the mass ratio of the hematoxylin to the absolute ethyl alcohol is 1 (75-85), the mass ratio of the hematoxylin to the glycerol is 1 (0.6-1), the mass ratio of the hematoxylin to the ascorbic acid is 1 (0.04-0.06), and the mass ratio of the hematoxylin to the polyethylene glycol 400 is 1 (2-3). Preferably, the preparation method of the modified hematoxylin b solution comprises the steps of adding deionized water into 30% ferric trichloride solution by mass fraction, uniformly stirring, then dropwise adding concentrated hydrochloric acid under continuous stirring, controlling the pH value to be 1.5-2.5, sequentially adding sodium citrate and aluminum potassium sulfate powder after the dropwise adding is completed, and continuously stirring until the solid is completely dissolved and the solution is clear, thus obtaining the modified hematoxylin b solution. Preferably, the mass ratio of the ferric trichloride solution to the deionized water is 1 (0.6-0.7), the mass ratio of the ferric trichloride solution to the concentrated hydrochloric acid is 1 (0.2-0.25), the mass ratio of the ferric trichloride solution to the sodium citrate is 1 (0.05-0.07), and the mass ratio of the ferric trichloride solution to the aluminum potassium sulfate powder is 1 (0.02-0.04). Preferably, the preparation method of the modified eosin dye solution comprises the steps of mixing 95-100 parts of acetic acid-sodium acetate buffer solution with the pH value of 4.8 with 1 part of water-soluble eosin Y powder, continuously stirring to form uniform red clear solution, sequentially adding 0.08-0.12 part of polysorbate-20 and 0.4-0.6 part of benzyl alcohol into the clear solution, continuously stirring until the additive components