RE: Need help with special processing...

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From:"Kellar, Eric" <> (by way of Marvin Hanna)
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Botanical microtechniques offer a bit of confusion for the beginner and the
experienced technogist. The solutions you have on hand for human histology
will suffice.

The first thing to consider is whether to demonstrate the cellular plant
detail or only the skeletal structures and the subsequent choices of
staining to be employed later.

To begin place a small portion of the living root, after thorough washings
in tap to remove soil or sand, into several changes of 95% ethanol for 48
hours. The 95% will become discolored. Then increase to 100% ethanol,
followed by xylene for a day each depending on size of specimen. Then
infiltrate in paraffin and section at 10 to 12 microns. Place sections onto
(+) treated slides. You may also try the processing on your automated
processor beginning in 95% ethanol.

If the skeletal structure is to be demonstrated only, then post-treat the
deparaffinized slides in a 1% aqueous solution of potassium hypochlorite and
check microscopically as the cellular constituents are leeched away. This is
known as skeletonizing plant tissue. After this is complete the sections
should be washed in several changes of DH20. The sections are then placed
into a 1% aqueous acetic acid solution for 5 minutes and then placed under
running tap.

An H&E would be nice for a student. Other more involved staining techniques
have been offered by Benda, Lands and Chamberlain and are still used today.
They involve aqueous solutions of light green, acid fuchsin, iodine green
and crystal violet to mention a few.

More information (on what exactly is to be demonstrated) would be helpful in
assisting you and the student further.

Eric C. Kellar
University of Pittsburgh Medical Center

-----Original Message-----
From: Maria Saenz
To: HistoNet Server
Sent: 1/23/00 7:25 AM
Subject: Need help with special processing...

We are in the process of helping out a doctor's son doing research and
been asked to process , cut, stain, etc his specimens.  The only thing
that I've never processed anything but human tissue.  The specimens we
be processing is of the plant kingdom variety....possibly radishes.  Can
anyone please us with this.  Please  include reagents of choice as well
suggested processing schedule.


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