Steve

You may find that your cell walls are autofluorescent even if not lignified.
There are a number of stains that will react with cellulose but in my experience
none of them are specific so you will get staining of cytoplasm and other cell
wall components as well. These include safranin, calcofluor (which needs UV
excitation), acriflavin, congo red (very weak - difficult to get staining). I
you want to specifically label cellulose its possible to label cellulase enzyme
with fluorescent dye and use that as a specific probe (sorry could not locate
the reference but it was published in the last 5 years). You can get protein
labelling kits from Molecular Probes and others such as Sigma, to make the
probe. If you don't care about cytoplasmic staining try acriflavin. It stains
lignin green and cellulose (ie. unlignified wall) red. You have to make the
stain fresh though - it looses its differential staining after a few days (use a
very dilute solution - should be pale yellow).
Hope that helps

Lloyd Donaldson
Forest Research, Rotorua, New Zealand
http://www.forestresearch.co.nz/mnz




Steve Taylor <[log in to unmask]> on 31/05/2000 19:31:38

Please respond to Confocal Microscopy List <[log in to unmask]>

To:   [log in to unmask]
cc:    (bcc: Lloyd Donaldson/NZ Forest Research Institute/NZ)

Subject:  Plant cell wall



Are there any recommendations out there for a fluorochrome suitable for
cellulose fibers in plant cell walls, alive and/or fixed?.

Steve Taylor

---------------------------------

Dr. S. Taylor
CLSM Unit
Institute of Biological Sciences
Cledwyn Building
University of Wales
Aberystwyth
Ceredigion
SY23 3DD
UK

[log in to unmask]
01970 621567
---------------------------------