Fwd: staining JB-4 sections

From:MaryLou

Techniques in Plastic for Light Microscopy
National Society for Histotechnology - Region IV Symposium
June 25, 1983   Ann Arbor, Michigan
Alexandra N. Brady, M.S., MT/HTL (ASCP)
Robert Schoonhoven, HTL (ASCP)

I like the Lee's Methylene Blue -Basic Fuchsin for an H&E look-a-like.
1) Stock methylene blue
              0.5 g methylene blue  (CI 52015)
              400 mL distilled water

2) Stock basic fuchsin
             0.5 g basic fuchsin    (CI 42510)
             400 mL distilled water

3) Stock phosphate buffer 0.1 M   pH 6.8
            49 mL  0.2 M sodium phosphate dibasic anhydrous (28.4 g Na2H2PO4 in 1 liter DH2O)
            51 mL  0.2 M  sodium phosphate monobasic monohydrate  (27.8 g NaH2PO4-H2O in 1 liter DH2O)
            100 mL distilled water

4)  95% ethyl alcohol

5)  Working methylene blue-basic fuchsin
            12.0 mL stock methylene blue
            12.0 mL stock basic fuchsin
            21.0 mL stock phosphate buffer
            15.0 mL 95% ethyl alcohol
                DISCARD AFTER USE

Procedure:
1)  Stain in working solution for 10 seconds

2)  Rinse in distilled water

3)  Dip in 95% alcohol quickly with agitation to remove background stain

4) Rinse in water

5) Dry and mount

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

Reference:  Sorvall Instruction Manual, Staining Procedures for Plastic Embedded Tissue, DuPont Company, Instrument Products, Biomedical Div.,  July, 1979

Date: Thu, 30 Jan 2003 16:01:38 +0100
X-PH: V4.1@mailhub3
From: =?iso-8859-2?Q?Michaela_Fr=FDzkov=E1?= <michaela.fryzkova@centrum.cz>
Subject: staining JB-4 sections
To: histonet <histonet@pathology.swmed.edu>

Dear Colleagues,
I need an advice about staining sections embedded in JB-4 Plus. I embed in
this medium and now I need stain my sections. I wanted to use regular
haematoxylin-eosin staining, but it doesn#180#t work. Do you know something
about that? Can you recommend me some other staining?

Thank you, very much.
Michaela Fryzkova

Department of Parasitology
Faculty of Science
Charles University
Prague
Czech Republic

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