Stains & Dyes

Popular Stains

Hematoxylin and Eosin (H&E)

This the basic morphology stain. that renders the nuclei purple, with all other tissue components 3 shades of pink.

Expected Results

Nuclei –Blue
Cytoplasm and other tissue elements – 3 Shades of Pink
Blood – Red

Breast H&E

Skin H&E

Brain H&E

Fish Eye H&E

Acid Fast Bacilli (AFB)

The counterstain used is by personal preference, we have used green or blue.

Expected Results

Acid-fast bacilli – Bright Red
Erythrocytes – Yellowish Orange
Other tissue elements – Pale blue

Alcian Blue and it’s Variations

These are great stains for a range of sulfated mucosubstances and mucins, glycosaminoglycans, hyaluronic acid, and sialomucins.

Expected Results

Weakly acidic sulfated Mucosubstances, hyaluronic acid and sialomucins- Dark blue
Background- Pink to red

Breast – Alcian Blue

Colon Alcian Blue / PAS

Colon Alcian blue variation staining strongly sulfated mucins

Colon Alcian Blue variation staining highly sulfated connective tissue

Alizarin Red S / Calcium

This stains for calcium. The minerals turn orange with a yellow background under a normal light microscope. Utilizing polarizing lenses makes the background turn dark blue or black with the minerals fluoresce orange.

Expected Results

Calcium Deposits (except oxalate) – Orange-Red
All other tissue elements – Yellow

Kidney stained with Alizarin Red S For Calcium

Polarized kidney stained with alizarin red S for Calcium deposits (orange)

Benholds Congo Red

This is a great stain for amyloidosis instead of doing IHC.

Expected Results

Amyloid Deposits – Red
Background and nuclei – Purple
Congo red -polarized

Rhodanine

Expected Results

Copper granules – Orange
Everything else – Blue

Field’s Stain

Expected results

Bacteria, including pylori – Deep Blue
Cellular elements – Similar to a Romanowsky stain
Malaria chromatin – Purple Red to Deep Red
Malaria cytoplasm – Blue

Fontana Masson

Expected results

Melanin – Black
Background – Pinkish red

May-Grunwald Giemsa

Expected results

Rickettsia – Violet
Nuclei – Blue
Cytoplasm, connective tissue – Pink
Erythrocytes – Salmon

Gram Brown and Hopps Method

Expected results

Gram Positive bacteria – Blue
Gram Negative bacteria – Red
Additional tissue elements –Yellow

Grocott’s Methenamine Silver (GMS)

Expected results

Fungi – Sharply delineated in Black
Pneumocystis carnii – Black
Mucin – Taupe to dark Grey
Inner parts of mycelia and hyphae – Old Rose
Background – Pale Green

Hall’s Method for Bile

Expected results

Bile (bilirubin)- Green
Connective Tissue – Red to pink
All other tissue – Yellow

Modification of Mowry’s 1958 Colloidal Iron Stain For Acid Mucopolysaccharides (Colloidal Iron)

Expected results

Acidic mucopolysaccharides and acidic epithelial mucins – Deep blue
Background – Red

Luxol Fast Blue (LFB) – We have 4 variations; LFB / H&E, LFB / PAS, LFB / NFR, LFB / CV

Expected results – LFB / H&E

Myelin- Blue
Nuclei- Purple
Background – Pink

LFB / H&E

Expected results – LFB / Periodic Acid Schiff (PAS)

Myelin- Blue
Nuclei- Purple
Background – Pink

LFB / PAS

Expected results – LFB / Cresyl Violet (CV)

Myelin – Blue
Nissel Substance – Violet
Nuclei – Violet

LFB / CV

Expected results – LFB / Nuclear Fast Red (NFR)

Myelin- Blue
Nucleus- Red

LFB / NFR

Mallory PTAH

Expected results

Cross-striations, fibrin – Blue
Nuclei – Blue
Collagen – Red-Brown

Heart PTAH

Masson’s Trichrome Blue or Green

Expected results

Nuclei- Black
Cytoplasm, keratin, muscle fiber and intercellular fibers- Red
Collagen – Blue or Green (depends on preference)

Masson’s Trichrome

Masson’s Trichrome – Colon

Masson’s Trichrome (green) Aorta

Masson’s Trichrome (green)

Mayer’s Mucicarmine

Expected results:

Mucin – Deep rose to red
Capsule of Cryptococcus – Deep rose to red
Nuclei – Black
Other tissue elements – Yellow

Movat’s Modified Pentachrome

Expected Results

Nuclei and elastic fibers—Black
Collagen—Yellow
Ground substance and mucin–Blue
Fibrinoid, fibrin–Intense Red

Oil Red O

Expected results

Fat Globules – Orange Red
Nuclei – Blue

Oil Red O – Liver

Orcein for Hepatitis

Expected results

HBsAg – Dark brown

Hepatitis B

Periodic Acid Schiff (PAS)

Expected results depend somewhat on the counterstain used:

Hematoxylin – Glycogen, mucin, fibrin, or thrombi, colloid droplets, hyaline of arteriosclerosis, hyaline deposits in glomeruli, granular cells in the renal arterioles where preserved, most basement membranes, colloid of pituitary stalks and thyroid, amyloid infiltration may show a positive reaction –Rose to Purplish Red

Nuclei – Blue
Light Green – Fungi – Red
Background – Pale Green

PAS With Digestion

Expected results

The PAS stain is used to detect containing a high proportion of carbohydrate macromolecules (glycogen, glycoprotein, proteoglycans), typically found in e.g. connective tissues, mucus, the glycocalyx, and basal laminae. http://(http://en.wikipedia.org/wiki/Periodic_acid%E2%80%93Schiff_stain)

The variation on the PAS technique involves simply exposing the section to the diastase enzyme amylase prior to continuing with the standard PAS method. The diagnostic purpose of performing the PASD technique includes:

The removal of glycogen to make it easier to identify mucins stained by the PAS stain.
Analysis of glycogen deposits within the liver.
Highlighting a thickened basement membrane for example in lupus.

https://skinpathonline.wordpress.com/2011/05/30/periodic-acid-schiff-diastase-pasd-special-stain-%E2%80%93-method-and-tips/

Picro Sirius Red and Fast Green

We have 2 variations of this stain.