Re: Histo in Plastic

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From:Mary Lou Norman <mlm11@cornell.edu>
To:Katie B <bresee98@yahoo.com>
Reply-To:
Date:Thu, 10 Jun 1999 10:08:25 -0400
Content-Type:text/plain; charset="us-ascii"


Katie,

This is my favorite:	 Apparently Lee got around!

Lee's Methylene Blue - Basic Fuchsin

1) Stock Methylene Blue
         0.5 g m.b. (CI 52015) 
         400 mL Distilled water  (DH2O)

2) Stock Basic Fuchsin
         0.5 g b.f.   (CI 42510)
         400 mL DH2O

3) Stock phosphate buffer 0.1 M pH 6.8
         49 Ml 0.2 M sodium phosphate dibasic anhydrous (28.4g Na2H2PO4 in
1000mL DH2O)
         51 Ml 0.2 M sodium phosphate monobasic monohydrate (27.6g
NaH2PO4'H2O in 1000mL DH2O)

4) 95% ethyl alcohol

5) Working m.b.-b.f.
          12.0 mL stock m.b.
          12.0 mL stock b.f.
          21.0 mL stock buffer
          15.0 mL 95% alcohol  (DISCARD AFTER USE)	

Procedure:

1)  Stain in working solution for 10 seconds

2)  Rinse in DH2O
 
3)  Dip in 95% quickly w/agitation to remove background staining

4)  Rinse in water

5)  Dry and mount

Results:     Nuclei- blue
             Cytoplasm, cilia, certain cell granules - shades of pink and red
             Cartilage and most cell granules - blue to purple

Ref:  Sorvall Instruction Manuel, Staining Procedures for Plastic Embedded
Tissue,
      Dupont, Co. Intruments Productsa, Biomedical Div., July, 1979

Where I got this:  Techniques in Plastic for Light Microscopy
                   NSH Region IV Symposium
                   6-25-83  Ann Harbor, Michigan                   
			Alexandra N. Brady, M.S., MT/HTL (ASCP)
                   Robert Schoonhoven, HTL (ASCP)

Phew!! 
                






At 12:57 09-06-1999 -0700, you wrote:
>Looking for a recipe for Methylene Blue-Basic Fuchsin stain to do in
>GMA embedded tissues.  My boss tells me it is the "old" way to do
>something like an H+E in plastic.
>
>Thanks!
>
>===
>Catherine "Katie" Bresee Bennett
>Laboratory for Experimental Pathology
>Department of Veterinary Pathology
>Michigan State University
>
>e-mail: bresee98@yahoo.com
>_________________________________________________________
>Do You Yahoo!?
>Get your free @yahoo.com address at http://mail.yahoo.com
>
>
>



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