Re: Acridine Orange and Calcofluor White Stains

From:"J. A. Kiernan"

Allison Scott wrote:
> Hello to all in histoland.  I'm in the need for the protocols for the
> acridine orange and calcofluor white stains.

Here's a calcofluor white method. It's for 2 um plastic
sections of plant tissue, dried onto slides. Should be
OK for paraffin, perhaps with minor modifications at
step 5.

1. Schiff's reagent, 2 hr.
2. Running tap water 5 min then dist water rinse.
3. Calcofluor white M2R (CI 40622) 10mg/100 ml, 2 min
4. Three rinses in dist water
5. Air dry, rinse in xylene, and apply coverslip,
   using a non-fluorescent mounting medium.

With broad-band UV excitation: Lignified cell walls  
brown, cellulose walls bright white.
The paper describing this (B.Mori & L.M.Bellani 1996
Biotech. Histochem. 71:71-72) includes two nice
big coloured micrographs.

John A. Kiernan
Department of Anatomy and Cell Biology
The University of Western Ontario
London,   Canada   N6A 5C1

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