Re: Ruthenium Red Protocol

From:"J. A. Kiernan" wrote:
> I am trying to identify components of wheat straw stems.  I want to look at
> the use of Ruthenium Red and whether this stain will help differentiate
> cells and other materials in harvested stems.  I have found a couple
> references that provide concentrations of RR to use but I do not have a
> complete prep-protocol for use of this stain.  Can anyone provide me with a
> staining procedure for ruthenium red and slide prep procedure?

You don't say whether it's for light or electron microscopy.
In "Plant Microtechnique and Microscopy" by Steven E. Ruzin
(Oxford univ Press, 1999) a variety of LM ruthenium red
are given, for staining pectin. The concentration, time, and
composition of the solution cannot be very critical. You should
be aware that this inorganic dye has no particular histochemical
specificity for pectin or anything else (see Pearse's
3rd edn, Vol 2). I've used it on paraffin sections a few times;
stained the same things as neutral red. 

Ruthenium red has many uses in electron microscopy, especially
contrast staining of skin (stratum corneum) and the glycocalyx, 
especially of bacteria. It also has some interesting
properties, interfering with calcium transport, neurogenic 
inflammation etc. See M. A. Hayat, 1993 Stains and Cytochemical 
Methods, p.305-318 for a review of all this, with practical 
instructions for EM applications.
John A. Kiernan
Department of Anatomy and Cell Biology
The University of Western Ontario
London,   Canada   N6A 5C1

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