Histologistics Stains List

Histologistics Inc A Histology Service

Our Stains

All of our stains are modified by in house specialists, who tailor them for bright, exact color staining for all types of tissues.  These stains are closely regulated by our trained staff through control tissues.  Staining solutions are made in house from powdered dyes to specific specifications and tested on control tissue before using.  All solutions are tested for correct pH, given lot numbers and expiration dates.  This monitoring system gives correct staining characteristics that are necessary for correct tissue identification.

This stains list is by no means the be all end all list for us, we are happy to work-up any particular stain that you prefer.  As we have stated in recent blog posts, we will also modify any of our stains to your liking.  One recent customer wanted nuclei present in their picrosirius red stain.  It took a couple trials to find out how our client wanted it to look, but they are now happy.  Most of our stains have been modified in some way for it’s users.  Our gram stain, Brown and Hopps, one client asked if we would not use the background stain because their computer program had trouble reading it.  Even though we had never done this, we tested it and found it to work just as well as the original stain.  It’s the little things in life that can make you happy, we are happy to help.

The stains below, and their pictures are done routinely at Histologistics.  These are only the stains and dyes.  We also routinely perform immunohistochemistry and immunofluorescence staining.  Since this is a very broad category and many of the pictures are subjective, posting them here is not suitable.    

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
microscopic breast stained with H&E
Breast H&E
Microscopic skin stained with H&E
Skin H&E
Microscopic brain stained with H&E
Brain H&E
microscopic fisheye stained with 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
microscopic picture of kidney with acid fast bacilli
Kidney with AFB
microscopic picture of kidney stained with AFB
Human Kidney AFB

 

 

 

 

Alcian Blue and it’s Variations + Alcian Blue / PAS

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
microscopic picture of breast stained with alcian blue
Breast – Alcian Blue
Microscopic picture of colon stained with alcian blue / PAS
Colon Alcian Blue / PAS
Microscopic picture of colon stained with a variation of alcian blue
Colon Alcian blue variation staining strongly sulfated mucins
Microscopic picture of colon stained with a variation of alcian blue
Colon Alcian Blue variation staining highly sulfated connective tissue

 

 

 

 

 

 

Alizarin Red S

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
microscopic picture of kidney stained with alizarin red S for calcium
Kidney stained with Alizarin Red S For Calcium
Microscopic picture of kidney stained with alizarin red S for Calcium
Polarized kidney stained with alizarin red S for Calcium deposits (orange)

 

 

 

 

 

 

Alkaline Congo Red

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

Expected Results

  • Amyloid Deposits – Red
  • Background and nuclei – Purple
Microscopic picture of kidney stained with congo red for amyloid
Kidney – Congo red for Amyloid Deposits (red)

 

 

 

 

 

Rhodanine

Expected Results

  • Copper granules – Orange
  • Everything else – Blue
Microscopic picture of liver with copper deposits stained with rhodanine
Liver with Copper deposits

 

 

 

 

 

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
microscopic picture of Intestine stained with Fields for helicobacter pylori
Intestine stained with Fields for helicobacter pylori

 

 

 

 

 

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
Microscopic picture of gram negative bacteria
Gram Negative Bacteria
Microscopic picture of Gram positive bacteria
Gram Positive Bacteria

 

 

 

 

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
Microscopic picture of Aspergillus fungus stained with Grocott's Methenamine Silver
Aspergillus fungus stained with Grocott’s Methenamine Silver
Microscopic picture of Pneumocystis Fungus - GMS
Pneumocystis Fungus – GMS
Microscopic picture of Blastomyces GMS
Blastomyces GMS

 

 

 

 

 

Hall’s Method for Bile

Expected results

  • Bile (bilirubin)- Green
  • Connective Tissue – Red to pink
  • All other tissue – Yellow
microscopic liver stained with Hall's method for Bile
Hall’s method for Bile (green) 40x
microscopic liver stained with Hall's method for Bile
Hall’s method for Bile (green) 100x

 

 

 

 

 

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
Microscopic picture of small intestine stained for colloidal iron
Colloidal Iron

 

 

 

 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
Microscopic picture of brain stained with LFB & H&E
LFB / H&E

 

 

 

 

Expected results – LFB / Periodic Acid Schiff (PAS)

  • Myelin- Blue
  • Nuclei- Purple
  • Background – Pink
microscopic picture of brain stained with LFB and periodic acid schiff
LFB / PAS

Expected results – LFB / Cresyl Violet (CV)

  • Myelin – Blue
  • Nissel Substance – Violet
  • Nuclei – Violet
Microscopic picture of brain stained with LFB cresyl violet
LFB / CV

 

 

 

 

Expected results – LFB / Nuclear Fast Red (NFR)

  • Myelin- Blue
  • Nucleus- Red
microscopic picture of brain stained with LFB and nuclear fast red
LFB / NFR

 

 

 

 

Mallory PTAH

Expected results

  • Cross-striations, fibrin – Blue
  • Nuclei – Blue
  • Collagen – Red-Brown
Microscopic picture of heart stained with PTAH
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)
Microscopic picture of heart stained with masson's trichrome
Masson’s Trichrome
microscopic picture of colon stained with Masson's trichrome
Masson’s Trichrome – Colon
microscopic picture of aorta stained with Masson's trichrome
Masson’s Trichrome (green) Aorta
microscopic picture of cord stained with Masson's trichrome
Masson’s Trichrome (green)

 

 

 

 

 

Movat’s Modified Pentachrome

Expected Results

  • Nuclei and elastic fibers—Black
  • Collagen—Yellow
  • Ground substance and mucin–Blue
  • Fibrinoid, fibrin–Intense Red
microscopic picture of colon stained with Movat's pentachrome
Colon Movat’s Pentachrome

 

 

 

 

 

Oil Red O

Expected results

  • Fat Globules – Orange Red
  • Nuclei – Blue
microscopic picture of liver stained with oil red O
Oil Red O – Liver

 

 

 

 

Orcein for Hepatitis

Expected results

  • HBsAg – Dark brown
microscopic picture of hepatitis stained with Orcein
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
microscopic picture of appendix stained with PAS for glycogen
PAS Appendix

 

 

 

 

 

 

Periodic Acid Schiff with Diastase (PASD)

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.

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

microscopic picture of appendix stained with PAS & PASD on liver
PAS & PASD on liver

 

 

 

 

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  
microscopic picture of polarized skin stained with picrosirius red
Skin Picrosirius Red (polarized)
microscopic picture of polarized skin stained with picrosirius red
Skin – picrosirius red

 

 

 

 

 

  2. One non-polarizing stain

Expected Results

  • Muscle – Red,
  • Cytoplasm –Yellow          
  • Collagen – Red  
  • Nuclei  – Black
microscopic heart stained with picrosirius red fast green
Heart – picrosirius red with nuclei

 

 

 

 

 

Pizzolato’s Method for Calcium Oxalate

Expected results

  • Calcium oxalate – Black
  • All other tissue components – Red
microscopic view of kidney with calcium deposits (black) stained with pizzoalto's
Kidney calcium (black) with Pizzolato’s

 

 

 

 

 

Prussian Blue for Ferric Iron

Expected results

  • Iron (ferric form) – Bright Blue Background – Depends on counterstain used
microscopic picti=ure of liver with iron deposits (blue)
Iron deposits (blue) – Liver

 

 

 

 

 

Safranin-O-Fast Green

Expected results:

  • Articular cartilage (proteoglycan) – Red
  • Nuclei- Blue
  • Background- Green
microscopic knee joint stained with safranin-o-fast green
Safranin-O Fast Green
microscopic knee joint stained with safranin-O
Joint Safranin-O

 

 

 

 

Snook’s Method for Reticulum

Expected results:

  • Reticulum fibers – lavender grey to black  
  • Background – pink to rose
microscopic picture of brain stained with snook's reticulin stain
Kidney reticulum (black) 20x
microscopic picture of liver with reticulin (black)
Liver Reticulin (black)

 

 

 

 

 

Toluidine Blue (T-Blue)

Expected results:

  • Histamine granules of the mast cell – Violet
  • Background and Nuclei – Blue
microscopic skin stained with toluidine blue, counterstained with fast green
Skin Toluidine Blue / fast green for Mast cells (purple)
microscopic skin stained with toluidine blue for mast cells
Human Skin Toluidine Blue, no counterstain, for Mast cells (purple)
microscopic bladder stained with toluidine blue for mast cells
Bladder – Toluidine Blue for Mast cells (purple)
microscopic heart stained with toluidine blue for mast cells
Heart – Toluidine Blue for Mast cells (purple)

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