Stains & Dyes

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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 (glycogenglycoproteinproteoglycans), typically found in e.g. connective tissuesmucus, the glycocalyx, and basal laminaehttp://(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:

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

 Picro Sirius Red and Fast Green

We have 2 variations of this stain.  

  1. The polarizing stain 

Expected Results

  • Muscle, cytoplasm –Yellow Collagen– Red
    • With polarizing microscopy:
  • Large collagen fibers–Yellow or Orange
  • Thin collagen fibers–Green  
Skin Picrosirius Red (polarized)
Skin – picrosirius red

  2. One non-polarizing stain

Expected Results

  • Muscle – Red,
  • Cytoplasm –Yellow          
  • Collagen – Red  
  • Nuclei  – Black
Heart – picrosirius red with nuclei
Human placenta picrosirius red fast green

Pizzolato’s Method for Calcium Oxalate

Expected results

  • Calcium oxalate – Black
  • All other tissue components – Red
Kidney calcium (black) with Pizzolato’s

Prussian Blue for Ferric Iron

Expected results

  • Iron (ferric form) – Bright Blue Background – Depends on counterstain used

Safranin-O-Fast Green

Expected results:

  • Articular cartilage (proteoglycan) – Red
  • Nuclei- Blue
  • Background- Green
Safranin-O Fast Green
Joint Safranin-O
Snook’s Method for Reticulum

Expected results:

  • Reticulum fibers – lavender grey to black  
  • Background – pink to rose
Kidney reticulum (black) 20x
Liver Reticulin (black)

Steiner & Steiner for Spirochetes

Expected results:

  • Spirochetes – Black
  • Background – Yellow
Human Lung with Spirochetes

Toluidine Blue (T-Blue)

Expected results:

  • Histamine granules of the mast cell – Violet
  • Background and Nuclei – Blue
Skin Toluidine Blue / fast green for Mast cells (purple)
Human Skin Toluidine Blue, no counterstain, for Mast cells (purple)
Bladder – Toluidine Blue for Mast cells (purple)
Heart – Toluidine Blue for Mast cells (purple)

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