[Histonet] Plant Tissue Processing

[John Kiernan]

Soft plant tissues are easily damaged before fixation. Cut (slice, not chop) with a very sharp razor blade, not scissors. The high water content can lead to collapse during dehydration, so this needs to be done gradually, especially through concentrations of alcohols greater than about 50%.  A formalin-acetic-alcohol fixative with 50% alcohol is a good fixative for plants. I have used Davidson's fixative, which contains somewhat less alcohol, for stems,  flowers and leaves, with good results. I've never tried making sections of grass.
 
There are several solvents and techniques for slowly and gently dehydrating and clearing plant and other "delicate" watery tissues. I strongly recommend "Botanical Microtechnique and Cytochemustry" by GP Berlyn & JP Miksche. Iowa State Univ Press, Ames IA (1976) ISBN 813802202. This 326-page hardcover book may still be available new, from the publisher, quite inexpensively. It gives very detailed descriptions of collection, fixation, processing, embedding and cutting sections (paraffin and plastic). A machine processing schedule for chunks of animal organs probably would be too harsh for non-woody plant material. 
 
John Kiernan
Anatomy, UWO
London, Canada
= = =
----- Original Message -----
From: Alan Bright <abright <@t> brightinstruments.com>
Date: Thursday, November 13, 2008 6:43
Subject: RE: [Histonet] Plant Tissue Processing
To: Peggy Bisher <mbisher <@t> Princeton.EDU>, Histonet <@t> lists.utsouthwestern.edu

> Dear Peggy,
> 
> I would have thought cryo sectioning would give much better 
> results. 
> 
> 
> Best Regards
> 
> Alan Bright
> 
> Bright Instrument Co.Ltd.
> St Margaret's Way
> Huntingdon
> Cambridgeshire
> PE29 6EU
> England
> 
> Tel No:+44 (0)1480 454528
> Fax No:+44 (0)1480 456031
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> 
> 
> 
> 
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Peggy
> Bisher
> Sent: 12 November 2008 17:02
> To: Histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] Plant Tissue Processing
> 
> I have a professor here who would like me to make 10-15 micron thick
> sections of lawn grass. I thought the best way to do this would be
> paraffin
> embedding. Does anybody have any suggestions as to the 
> time/types for my
> paraffin tissue processor? I am not very familiar working with plant
> material.
> 
> Thank you very much, Maggie
> 
> 
> Margaret E. Bisher
> Electron Microscopy & Histology Core Facility Manager
> Department of Molecular Biology
> Princeton University
> Moffett Laboratory, Room 113
> Princeton, New Jersey
> Office: (609) 258-7026
> Fax: (609) 258-8468
> mbisher <@t> princeton.edu
> 
> 
> 
> 
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