Other Staining solution, Lysochromes, and Chief solvents used for stains

Van Gieson Stain

• “Acid Fuchsin-Picric Acid”

• This uses acid fuchsin and picric acid to differentiate connected tissues in histological samples.

Alcian Blue

• A complex water-soluble phthalocyanin dye similar to chlorophyll, which stains acid mucopolysaccharides by forming salt linkages with them.

• It is also an excellent stain because it is simple, it produces a striking blue color, and it is resistant to various counter-staining procedures.

• It is more specific for connective tissue and epithelial mucin due to its use as an acid solution.

Benzidine

A staining agent used specifically for hemoglobin, allowing for the visualization of blood components in histological samples.

Celestine Blue

• An oxazine dye used as an alternative to iron hematoxylin nuclear stain, producing a strong and precise nuclear stain that is resistant to decolorization by succeeding acid stains and solutions.

• It is also resistant to strong acid dyes, and is recommended for routine staining of fixed sections, giving a good nuclear definition when used in conjunction with alum hematoxylin

Cresyl Violet

• Cresyl violet is commonly used in histology to stain nervous tissues.

• It stains the acidic components of neuronal cytoplasm, specifically nissl bodies, a violet color.

Iodine

• is one of the oldest stains used in microscopy, primarily for studying starch granules.

• It stains amyloid, cellulose, starch, carotene, and glycogen, making it useful in histological and biochemical applications.

• is used to remove mercuric fixative artifact pigments and serves as a reagent to alter crystal and methyl violet dyes.

Methyl Green

• stains chromatin green in the presence of an acid.

• It gives false positive reactions with certain secretions, such as mucin.

• Methyl green is used commonly with bright field microscopes to dye the chromatin of cells so that they are more easily viewed.

Oil Red O

• A dye that is more soluble in fat than in water or alcohols, hence it is used as a stain for neutral lipids.

• It can identify neutral lipids and fatty acids in smears and tissues.

• It is a rapid and a simple stain.

• It can also be useful in identifying fat emboli in lung tissue or clot sections of peripheral blood.

Periodic Acid Schiff

• An all-around useful stain for many things.

• It stains glycogen, mucin, mucoprotein, glycoprotein, basement membranes, capsules, and blood vessels as well as fungi and intracellular carbohydrates.

• Self-desecrate mucus are strongly stained.

• A pre-digestion step with amylase will remove staining for glycogen, which works by periodic acid oxidation of hydroxyl groups into aldehydes, which react with the Schiff reagent to produce a reddish-purple color.

Silver Nitrate

Used in 10% aqueous solution to prepare various dilutions to be used in identification of spirochetes, reticulum, and other fiber stains.

Victoria Blue

used to demonstrate neuroglia in frozen sections.

Acrdine Orange

• A basic acridine fluorochrome, which permits discrimination between dead and living cells, giving green fluorescence for DNA and a red fluorescence for RNA.

• It is also a nucleic acid selective fluorescent cationic dye, useful for cell cycle determination.

• it is also useful as a non specific stain for backlighting conventionally stained cells on the surface of a solid sample of tissue

Aniline Blue

• It is a cytoplasmic stain used for counter-staining of epithelial sections.

• It is composed of aniline blue + distilled water + glacial acetic acid.

Bismark Brown

• Serves as a contrast stain in various techniques, including gram staining, the acid-fast method, and the Papanicolaou technique.

• Additionally, it is used to stain diphtheria organisms, aiding in their identification under the microscope.

Congo Red

• Best known as an indicator, but may be utilized as a stain for axis cylinders in embryos.

• It is also used as a 4% aqueous solution in staining elastic tissues, amyloid, and myelin, using Krajian’s method.

Giemsa Stain

• This consists of a mixture of methylene blue and eosin, and it is used for staining blood to differentiate leukocytes.

• It is mostly used on methanol-fixed blood films, where it stains erythrocytes pink and the different types of leukocytes, allowing their identification according to size and shape of their nucleus.

Janus Green B

It is used for demonstrating mitochondria during intravital staining.

Methylene Blue

• It is a common basic nuclear stain employed with eosin to provide marked differentiation of various structures in the tissue.

• stains acidic cell parts like nucleus and is a good counterstain with Eosin Y.

• It is also a valuable stain for plasma cells and may also be employed in cytological examinations of fresh sputum for malignant cells.

• It is also used as a bacterial stain for evaluation and differentiation of bacterial organisms, also for diagnosis of diphtheria and for vital staining of the nervous tissue.

Orcein

It is an excellent stain for elastic fibers and is especially recommended in dermatological studies due to its ability to demonstrate the finest and most delicate fibers in the skin.

Prussian Blue

• is an insoluble colored salt of ferric ferrocyanide and iron cyanide compound.

• It is normally utilized for the manufacture of paints, but may be used for micro-anatomical color contrast of specimens and for demonstration of the blood and lymph vessels by injection or intravital staining.

Safranin O

• is a nuclear stain.

• It produces red nuclei and is used primarily as a counterstain.

• Safranin may also be used to give a yellow color to collagen.

Wright Stain

This is primarily used for blood cell staining with four major staining properties.

Acrdine Red 3B

is used to demonstrate deposits of calcium salts and possible sites of phosphatase activities.

Basic Fuchsin

• A plasma stain utilized for deep staining of acid-fast organisms, for mitochondria, for differentiation of smooth muscles with the use of picric acid.

• It is also a main constituent of Feulgen’s and Schiff's reagent for the detection of aldehydes.

• It is also a constituent of Van Gieson solution for connective tissues, mucin, and for elastic tissue staining.

Carmine

• It is used as a chromatin stain for fresh materials in smear preparations.

• It is slightly soluble in water at a neutral reaction, and usually kept in ammonical solution, which changes its properties due to oxidation.

• It is usually combined with aluminum chloride to stain glycogen, and it is now called best carmine solution.

Crystal Violet

• A nuclear or chromatin stain used for staining amyloid in frozen sections and platelets in blood.

• Gentian violet is the staining solution formed by the mixture of crystal violet, methyl violet, and dexterin.

Gold Sublimate

is the stain used for metallic impregnation, made up of gold chloride and mercuric chloride.

Malachite Green

• A weakly basic dye used as a contrast stain for staining ascaris eggs and erythrocytes, and as a bacterial spore stain.

• It is also used as both a decolorizer and a counterstain.

Nile Red

• Nile Blue Oxazone

• It is formed by boiling nile blue with sulfuric acid.

• This produces a mix of nile red and nile blue.

• is a lipophilic stain.

• It will also accumulate in lipid globules inside cells.

• It fluoresces strongly when partitioned into lipids.

Osmium Tetroxide

• is a selective stain for unsaturated lipids and for lipoproteins such as myelin, which it stains black.

• Making it useful in lipid histochemistry and electron microscopy.

Rhodamine B

is used with osmic acid to fix and stain blood and granular tissues.

Toluidine Blue

• A nuclear stain also for fixed tissues, used as a substitute for thionine in fresh frozen tissue sections.

• It is recommended for staining of nasal granules or chromophilic bodies.

• It is a particularly versatile dye that stains nuclear blue and can be used to differentiate different types of granules, including mast cells.

Acid fuchsin (Masson stain)

may be used to stain collagen, smooth muscle, or mitochondria

Gram’s iodine

plays a crucial role in bacterial identification and differentiation

Lugol's iodine

reacts with starches, turning block upon contact.

Basophilia

It has affinity for methylene blue, which stains basophils and nucleic acids

Azurophilia

It has affinity for oxidized methylene blue.

Acidophilia

It stains azurophilic granules.

Neutrophilia

It has affinity for dye complexes, staining neutrophils.

Lysochromes or oil-soluble dyes

• are not real dyes, because they do not have their auxochrome groups.

• They give color to lipids simply because they are more soluble in lipid medium of the tissues than in their medium of 70% alcohol.

• This is used for the demonstration of intracellular fats, which are colored black, orange, and red, respectively.

Sudan black

• It is a stain that colors fat droplets black and is the most sensitive of the oil-soluble dyes.

• It possesses two secondary amino groups per molecule, making it a slightly basic dye which may cause nonspecific staining.

• It is prepared as a 0.5% solution boiled in 70% ethanol for 10 minutes under a reflux condenser.

• It is a very unstable solution and should be discarded if the usual blue to black color turns to a brownish black.

• stains phospholipids as well as neutral fats.

Sudan IV

• Scharlach R

• it has no secondary amino group and it does not color phospholipids or the fine lipid droplets.

• It is prepared by saturating the dye in one part of 2% benzoic acid in a 70% alcohol and one part of acetone, forming a very stable solution which may be used repeatedly as long as it is filtered.

• Addition of benzoic acid intensifies fat and prevents rapid deterioration of the solutions.

• It is recommended for staining triglycerides or neutral lipids, giving them a deep and intense red stain.

Sudan III

• It was the first Sudan dye to be introduced into histochemistry.

• It is also a fat soluble and is a good as a fat stain for central nervous system tissues, giving a less deep and lighter orange stain compared to the darker staining in Sudan IV.

TRUE

TRUE or FALSE: The chief solvents used for stains play a crucial role in preparing and dissolving dyes for histological applications.

Water

The most commonly used solvent and should always be distilled to ensure purity unless specified otherwise

Alcohol

• Particularly ethyl alcohol, is utilized in various concentrations depending on the staining requirements

• Methyl alcohol, usually in the absolute form, is primarily used in blood stain preparation and must be acetone free to prevent unwanted chemical interactions

Aniline Water

Prepared by adding 10 ml of aniline to every 0.5 to 1 liter of hot distilled water, then shaking, cooling, and filtering the solution to ensure clarity

Phenol

Used in aqueous solutions ranging from 0.5 to 5%, serving as a solvent that enhances the solubility and penetration of certain dyes into tissues.