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
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
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
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
Expected results – LFB / Periodic Acid Schiff (PAS)
- Myelin- Blue
- Nuclei- Purple
- Background – Pink
Expected results – LFB / Cresyl Violet (CV)
- Myelin – Blue
- Nissel Substance – Violet
- Nuclei – Violet
Expected results – LFB / Nuclear Fast Red (NFR)
- Myelin- Blue
- Nucleus- Red
Mallory PTAH
Expected results
- Cross-striations, fibrin – Blue
- Nuclei – Blue
- Collagen – Red-Brown
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)
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
Orcein for Hepatitis
Expected results
- HBsAg – Dark brown
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.
Picro Sirius Red and Fast Green
We have 2 variations of this stain.
- The polarizing stain
Expected Results
- Muscle, cytoplasm –Yellow Collagen– Red
- With polarizing microscopy:
- Large collagen fibers–Yellow or Orange
- Thin collagen fibers–Green
2. One non-polarizing stain
Expected Results
- Muscle – Red,
- Cytoplasm –Yellow
- Collagen – Red
- Nuclei – Black
Pizzolato’s Method for Calcium Oxalate
Expected results
- Calcium oxalate – Black
- All other tissue components – Red
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
Snook’s Method for Reticulum
Expected results:
- Reticulum fibers – lavender grey to black
- Background – pink to rose
Steiner & Steiner for Spirochetes
Expected results:
- Spirochetes – Black
- Background – Yellow
Toluidine Blue (T-Blue)
Expected results:
- Histamine granules of the mast cell – Violet
- Background and Nuclei – Blue
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