Histology Special Stain Protocols


All the protocols I used at my company were tested and modified by us for bright contrasting staining. I will try to post all my special staining protocols here. I’m working to modify them one by one to include the solutions used. We did not buy pre-made kits, we bought the individual chemicals and made our own staining solutions, with exceptions like hematoxylin, Bouin’s Fixative.

Human Stained with Acid Fast Bacilli (AFB)
Rat Colon Stained with Alcian Blue PAS
Human Kidney Stained with Alizarin Red S for Calcium
Beilschowsky Special Stain on Mouse Brain For Neurofibrils and Senile Plaques
Human Umbilical Cord Stained with Brown and Brenn Gram for bacteria (gram positive blue) – 1
Human Placenta – Brown and Hopps Gram Stain
Human Kidney Stained with Congo Red for Amyloid Amyloid
Human Lung Stained with Fite for Nocardia
Human Appendix Stained with Fontana Masson for Melanin
Human Kidney Stained with Gomori’s Periodic Acid Methanamine-Silver for Basement Membrane
Human Intestine Stained with Grocott’s Methenamine Silver for Fungi
Human Bone Marrow Stained with Hematoxylin & Eosin (H&E) 1
Mouse Heart Stained with Hematoxylin and Eosin (H&E)
Human Liver Stained with Halls for Bile (Green)
Human Liver Stained with Jones for Reticulin (black)
Rat Intestine Stained with Luna’s Method for Eosinophils and Erythrocytes 20x
Rat Brain Stained with Luxol Fast Blue + Hematoxylin & Eosin (LFBH&E) for Myelin Sheath (Blue) and Nuclei (Purple)
Rat Brain Stained with Luxol Fast Blue/ Nuclear Fast Red (LFBNFR) For Myelin Sheath
Rat Brain Stained with Luxol Fast Blue Cresyl Violet (LFBCV) for Myelin Sheath (Blue) and Nissl (Purple)
Rat Heart Stained with PTAH for Muscle Striations
Canine Heart Stained with Masson’s Trichrome
Human Small intestine Stained with Mayer’s Mucicarmine for Glycosaminoglycans
Human Spleen Stained with May-Grunwald Giemsa
Human Colon Stained with Mowry’s for Colloidal Iron (Blue)
Mouse liver Stained with Oil Red O for lipids (red)
Human Liver Stained with Orcein for Hepatitis
Rat Colon Stained with Periodic Acid Schiff / Alcian Blue (PASAB)
Human Liver Stained with PASD and PAS for a Comparison
Human Liver Stained with Pearls Prussian Blue for Ferric Iron (Blue)
Human Kidney Stained with Pizzolato’s for Calcium (Black)
Human Liver Stained with Rhodanine for Copper (Orange)
Human Skin Stained with Movats Modified Pentachrome
Mouse Skin Stained with Picrosirius Red for Collagen (Red)
Human Liver Stained with Sirius Red/Green Polarized for Collagen
Rat Muscle Stained with Picro-Sirius-Green/Red for Collagen (Red)
Rat Knee Joint Stained with Safranin-O Fast Green for Articular Cartilage (red)
Human Kidney Stained with Snook’s Method for Reticulum (Black)
Human Lung Stained with Steiner and Steiner for Spirochetes (Black)
Human Skin Stained with Toluidine Blue (T-blue) for Mast Cells (Purple)
Human Appendix Stained with Van Gieson Stain for Muscle (Yellow) Collagen (Red)
Porcine Aorta Stained with Verhoef Van Gieson stain for Elastic Fibers (Black)
Human Kidney Stained with Von Kossa for Calcium Salts (Black)

Are there more stains than this? Absolutely, yes, tons more stains and protocols. Most of these protocols were originally in histology books that we tested and modified for our use.

If you need a different protocol than found here, I suggest you look in a histology book like Theory and Practice of Histotechnology by Sheehan and Hrapchak, or Manual of Histologic Staining Methods by Lee Luna.

If these are not available to you, then try this website StainsFile.

The University of Utah is a great resource for all things histology/pathology.

If you buy special stain kits (pre-made) most come with staining protocols. I do suggest trying any new protocol on control tissues before using it on study tissues. You want to make sure the stain is perfect before staining research or clinical material.