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Comprehensive Guide to Staining Techniques

This document describes 31 different stains used for various tissue staining purposes. Some key stains mentioned are Van Gieson stain for connective tissues, Acridine Orange for distinguishing living and dead cells, Alcian Blue for acid mucopolysaccharides, Methylene Blue as a basic nuclear stain, Toluidine Blue as a substitute for thionine in fresh frozen sections, Giemsa Stain for staining blood cells, Orcein for elastic fibers, and Picric Acid as a contrast stain with acid fuchsin for connective tissue staining. The stains described are used to stain a variety of cellular structures and components for microscopic examination.

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Mdrrmo Maconacon
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476 views3 pages

Comprehensive Guide to Staining Techniques

This document describes 31 different stains used for various tissue staining purposes. Some key stains mentioned are Van Gieson stain for connective tissues, Acridine Orange for distinguishing living and dead cells, Alcian Blue for acid mucopolysaccharides, Methylene Blue as a basic nuclear stain, Toluidine Blue as a substitute for thionine in fresh frozen sections, Giemsa Stain for staining blood cells, Orcein for elastic fibers, and Picric Acid as a contrast stain with acid fuchsin for connective tissue staining. The stains described are used to stain a variety of cellular structures and components for microscopic examination.

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Mdrrmo Maconacon
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  • Celestine Blue: Serves as a substitute in hematoxylin staining procedures.
  • Toluidine Blue: Used in microanatomical and circulation system studies.
  • Acriflavine: Used for demonstrating phosphate activities in cells.
  • Van Gieson stain (Acid Fuchsin-Picric Acid): Describes the use of Van Gieson stain for demonstrating connective tissues.
  • Acridine Orange: Explains use for genetic material fluorescence and monitoring cellular processes.
  • Congo Red: Stain used particularly for cylinders in nephrology research.
  • Alcian Blue: Highlights acid mucopolysaccharides in biological samples.
  • Basic Fuchsin: Used in fast staining of mycobacteria and fungus.
  • Bismarck Brown: Serves as a background stain for counterstaining processes.
  • Iodine: Common stain for starch in histological applications.
  • Benzidine Brown: Used for staining cellulose in plant structures.
  • Victoria Blue: Stains neuroglia in frozen sections and chromosomes.
  • Coomassie Blue: Commonly used in protein band visualization after electrophoresis.
  • Giemsa Stain: Used for differential staining in hematology, primarily for blood smears.
  • Carmine: A traditional stain for identifying glycogen deposits.
  • Mayer's Carmalum: Alternative to carmine for staining glycogen with trichrome protocols.
  • Gold Sublimate: Used in histology for intricate tissue detail techniques.
  • Janus Green B: Serves in demonstrating mitochondria visually in cells.
  • Orcein: Highlights elastic fibers in dermatological sections.
  • Osmium Tetroxide: Primarily a fixative for lipid components within cells, stains fat black.
  • Lead Hematoxylin: Used for nuclear details in advanced tissue preparations.
  • Silver Nitrate: 0.1% aqueous solution used for biological stain preparations.
  • Picric Acid: Used for contrast in acid fuchsin methodologies.

OTHER STAINS

(Gregorios)

1. Van Gieson stain (Acid Fuchsin-Picric Acid)


 for demonstration of connective tissues.
 Weakens after long standing (Remedy: add few drops of fresh acid fuchsin)
2. Acridine Orange
 Basic acridine fluorochrome
 Permits discrimination between dead and living cells.
 Gives green fluorescence for DNA, red fluorescence for RNA.
3. Acridine Red 3B
 Demonstrates deposits of calcium salts and possible sites of phosphate activities.
4. Congo Red
 Indicatorl stain for axis cylinders in embryos
 Used as 4% aqueous solution in Kraijan’s method of staining elastic tissues, amyloid, and myeloid.
5. Neutral Red
 Basic dye; for observing cell granules and vacuoles of phagocytic cell.
6. Alcian Blue
 Water-soluble, pthalocyanin dye, similar to chlorophyll.
 Stains acid mucopolysaccharides by forming salt linkages.
 Produces STRIKING BLUE COLOR
 Resistant to various counterstaining procedure
 More specific for CT and epithelial mucin.
7. Aniline Blue
 Cytoplasmic stain used for counterstaining epithelial mucins.
8. Celestine Blue
 Resistant to strong acid dyes
 For routing staining of fixed sections, giving a good nuclear definition when used in conjunction with
alum hematoxylin.
9. Methylene Blue
 Basic nuclear stain employed with eosin.
 Polychroming – oxidation of methylene blue; resulting mixture of methylene blue, azure, thazoles is
Polychrome Methylene Blue.
 Nuclei Blue – cartilage, matrix, mucin, mast cell granules; Reddish-Violet – connective tissues
 Valuable stain for plasma cells; also for cytological exam of fresh sputum for malignant cells.
 Bacterial stain for bacterial organisms; for diagnosis of diptheria; vital stain of nervous tissue.
10. Night Blue
 Substiture for carbol fuchsin in acid fast staining.
11. Prussian Blue
 Manufacture of paints; may be used for microanatomical contrast of specimen.
 Demonstration of circulatory system by injection.

12. Toluidine Blue

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 Nuclear stain for fixed tissues
 Used as substiture for thionine in fresh frozen tissue sections
 For staining Nissl granules or chromophilic bodies
13. Victoria Blue
 Neuroglia in frozen sections
14. Basic Fuchsin
 Deep staining for acid fast organisms
 For mitochondria, differentiation of smooth muscles with the use of picric acid.
 Main constituent of Feulgen’s and Schiff’s reagent.
 For the detection of aldehydes of Van Gieson’s solution for connective tissues, mucin, elastic
tissue.
ex: Carbol Fuchsin, Coleman’s Feulgen’s reagent, Schiffs rgt, Mallory’s Fuchsin stain, Aldehyde
Fuchsin (Gomori’s stain)
15. Benzidine
 Staining hemoglobin
16. Bismarck Brown
 Contrast stain for gram’s technique in acid fast and papanicolau method
 For staining diphtheria organisms
17. Carmine
 Chromatin stain for fresh materials in smear preparation.
 Usually combined with aluminum chloride to stain glycogen (Best Carmine Stain)
18. Mayer’s Carmalum Solution
 Mordanted dye acting as a basic dye and staining acidic substances.
19. Crystal Violet
 Nuclear or chromatin stain used for amyloid in frozen sections and platelets in blood.
 Gentian Violet – mixture of crystal violet, methyl violet, dextrin.
20. Methylene Violet
 Metachromatic dye for coloring nuclei of leukocytes reddish-purple in presence of methylene blue.
21. Giemsa Stain
 Staining blood to different leukocytes
22. Gold Sublimate
 Used for metallic impregnation, made up of gold chloride and mercuric chloride
23. Iodine
 Oldest of all stains; for starch granules
 Stains amyloid, cellulose, starch, carotene, glycogen
 For removal of mercuric fixative artefact pigments.
24. Janus Green B
 Mitochondria during intravital staining
25. Malachite Green
 Contrast stain for ascaris eggs and erythrocytes
 Bacterial spore stain; used as a decolorizer and counterstain
26. Methyl Green
 Stains chromatin green; false positive reaction with mucin.

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27. Orcein
 For elastic fibers
 For dermatological studies due to its ability to demonstrate the most delicate fibers in skin.
28. Osmium tetroxide
 Fixative; used to stain fat
 Prevents specific staining of lipids
 Fat – reduces it to osmium dioxide; stained BLACK
29. Picric acid
 Contrast stain to acid fuchsin, for demonstration of connective tissue (Van Gieson stain)
 Cytoplasmic stain in contrast to basic dyes
 Counterstain to crystal violet; fixative and decalcifying agent
30. Rhodamine B
 Used with osmic acid to fix and stain blood and glandular tissues.
31. Silver Nitrate
 Used as 10% aqueous solution to prepare various dilutions to be used in identification of
spirochetes, reticulum and other fiber stains.

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