Connective & Muscle Tissue

Staining Mechanisms

100

Oil Red O utalizes this staining mechanism

Physical

200

Mast cells stained with toluidine blue are said to stain by this staining mechanism

Metachromatic Staining (Mast cells stain purple while the dye solution and other tissue elements are blue or orthochromatic)

300

Mallory PTAH is this type of staining solution

Polychromatic (1 solution that gives 2 major colors)

400

Alkaline alcohol converts Alcian blue into this insoluble pigment in the Movat Pentachrome procedure

Monastral fast blue

500

This type of tissue/reaction (ex: Reticulin fibers) has the ability to adsorb silver from solution, but requires a reducing agent to reduce the silver to a visible metallic form

Argyrophilic

Staining Solutions

100

Chromotrope 2R and Phosphotungstic Acid stain these tissues red in the Gomori 1 Step Trichrome

Muscle and Cytoplasm

200

Picric acid and Acid fuchsin are components of this staining solution

Van Gieson

300

When using this solution to stain basement membranes, the endpoint should be determined by the glomerular basement membranes rather than the tubular membranes. Slides should be checked frequently in this solution as optimum staining intensity is difficult to achieve. 

Methenamine Silver Solution

400

Iron hematoxylin is used rather than aluminum hematoxylin for nuclear staining in Trichrome methods for this reason

Subsequent acid solutions would decolorize aluminum hematoxylin stained nuclei

500

These are the 5 dyes of the Russel-Movat Pentachrome stain

Alcian blue: Acidic mucosubstances and ground substance

Iron hematoxylin: Elastic fibers and Nuclei

Crocein scarlet and acid fuchsin: Muscle and Cell cytoplasm

Alcoholic Safran: Collagen

What's Your Function?

100

This is the toner used in most silver staining methods

Gold chloride

200

Fat is chemically fixed and maintained through routine processing in tissue by this solution

Osmium tetroxide

300

This is the role of Phophotungstic and/or phosphomolybdic acid in the Masson Trichrome method

Removes Biebrich scarlet from collagen so that it can be counterstained

400

Ferric chloride performs these functions in the Verhoeff Elastic stain

Mordant and Differentiator (excess mordant). Also helps oxidize hematoxylin to hematein.

500

This can ripen Mallory PTAH for immediate use

Potassium permanganate

Trouble Shooting

100

This is how to remove cloudiness that may appear on slides counterstained with nuclear fast red

Back up to the tap water rinse then continue dehydration and clearing steps

200

This is the result of using metal forceps or glassware that wasn't chemically cleaned during silver staining methods

Non-specific silver staining and/or precipitation

300

This may result in collagen that is the same color as cytoplasm and muscle in the van Gieson picric acid-acid fuchsin stain

Not using saturated picric acid

400

This may cause faded blue staining of connective tissue in the Masson Trichrome stain

Over-differentiation in acetic acid solution or collagen that is pathologically altered

500

In the Movat Pentachrome procedure this may inhibit staining after the Alcian blue step

Incomplete removal of alkaline alcohol during the tap water rinse

Potpourri

100

Sections to demonstrate glomerular basement membranes should be this thickness

2 microns

200

This is how preparing tissues for Oil Red O differs from routine paraffin processing/microtomy

Frozen tissues are embedded in OCT (Optimal Cutting Temperature media) and sectioned with a cryostat

300

This is the preferred fixative for Masson Trichrome or mordant for formalin fixed tissues

Bouin solution

400

In this stain the differentiation step is very critical and can be variable from slide to slide. It is recommended to differentiate each slide individually.

Verhoeff Elastic Stain (Verhoeff-van Gieson)

500

Silver impregnation stains for reticulin depend on the formation of this chemical group

Aldehydes (through oxidation of carbohydrates)