[Histonet] Convallaria root section staining method

[John Kiernan]

Kasten, FH. Ch2 in Clark, G ed. 1981. Staining Procedures, 4 th ed. Baltimore: Williams & Wilkins, for the Biological Stain Commission. pp.57-58. Has extensive discussion and several references.
SE Ruzin (1999) Plant Microtechnique and Microscopy. New York: Oxford Univ. Press (ISBN: 0195089561), p.161-162. Recommended a much more dilute working stain solution.

Solutions.
    Acridine orange (CI 46005)  stock solution (100mg in 100ml water).
     0.1M phosphate buffer, pH6.0.
     Acridine orange working solution: Stock sol'n diluted 1:10 with buffer.

Method.
    Dewax and hydrate sections.
    Buffer solution, 2 min.
    Stain in working acridine orange solution, 15 min.
    Drain onto absorbent paper and rinse in buffer, 2 X 2 min.
    Mount in buffer; ring coverslip with wax or nail polish.
    Examine and photograph immediately.
        Excitation 487nm; Emission 520 nm.

Preparations are not permanent but slides can be re-stained.

Hope this helps. I've never done it myself.
John Kiernan
= = =
________________________________
From: Carl Hobbs via Histonet <histonet at lists.utsouthwestern.edu>
Sent: December 11, 2024 4:24 AM
To: histonet <histonet at lists.utsouthwestern.edu>
Subject: [Histonet] Convallaria root section staining method

Hi
I have prepared Pwax sections of convallaria root.
I want to use them as positive controls for fluorescence microscopy
( my Zeiss one was smashed)
I will be most grateful if anyone has an Acridine orange staining method to share?

Kind regards

Carl


Carl Hobbs FIBMS
Histology and Imaging Manager
Wolfson SPaRC
Guys Campus, London Bridge
Kings College London
London
SE1 1UL

020 7848 6810
_______________________________________________
Histonet mailing list
Histonet at lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet