[Histonet] whipf's polychrome procedure

James L Burchette burch007 <@t> mc.duke.edu
Tue Aug 11 10:30:30 CDT 2009

Here is  the polychrome formula I use.

Polychrome Methylene Blue Stain

Tissue and Fixation
Frozen sections and formalin-fixed, paraffin embedded

Rapid differential staining of tissue sections. Metachromatic stain.

Stock solutions:
A.      12% aqueous potassium carbonate, 100 ml.
B.      1% aqueous methylene blue, 4000 ml.
C.      10% aqueous glacial acetic acid, 50 ml.

Staining solutions:
1.      Mix 100 ml of A with 5000 ml of B.
2.      Gently boil for 60 minutes. Cool to room temperature.
3.      Add 50 ml of C.
4.      Store in a loosely capped bottle for further oxidation.

1.      Dip frozen sections in stain for 5 seconds.
2.      Rinse excess stain from slide.
3.      Mount in water with a cover glass if desired.

Nuclei blue, cytoplasm, muscle, connective tissue, pink to purple.

The sections are not permanent.
Solution works best after aging for at least 12 months, recognizable by a 
rich tinctoral quality.
Increase stock solution volumes to make a larger quantity of working 
stain. Filter before use.

Reference: Terry, BT, J. Lab and Clinical Medicine, Vol. 13, page 550, 

"Woodward, Denise" <denise.woodward <@t> uconn.edu> 
Sent by: histonet-bounces <@t> lists.utsouthwestern.edu
08/11/2009 11:23 AM

"histonet <@t> lists.utsouthwestern.edu" <histonet <@t> lists.utsouthwestern.edu>

[Histonet] whipf's polychrome procedure

Hello all,
One of our veterinary pathology residents from France is looking for a 
staining procedure for "Whipf's Polychrome".
Anyone have it and willing to share?
Denise Long Woodward
University of Connecticut
Connecticut Veterinary Medical Diagnostic Laboratory
Dept. of Pathobiology and Veterinary Sciences
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