Leaf clearing techniques?

Karen Snetselaar ksnetsel at sju.edu
Fri Jul 22 15:24:13 EST 1994

Biomancer (barosb at alleg.edu) wrote:
: I'm doing research with epidermal and mesophyll cells, especially  
: counting cells per unit area after ABA treatment in Hippuris  
: vulgaris...does anybody have a good, quick clearing method that will   
: remove everything excpet cell walls?  I've  been using 90% EtOH followed  
: by 7% NaOH, but it doesn't clear enough that I can see mesophyll cell  
: walls.  Any suggestions?  Most of the readings that I've looked at are  
: for the staining of xylem, but I don't need to see the xylem, and some  
: even degrade cellulose, which for me would be bad.

Place the tissue into a saturated solution of chloral hydrate (250g/100ml)
after you rinse out the NaOH.  Most leaves clear after a few days.  If you
have access to a microscope with Differential Interference Microscopy, you
will be able to see your cells fine without any stains.  If you need to
stain the leaves, you can find recipes and instructions in: Berlyn, G. P.,
and Miksche, J. P.  Botanical microtechnique and cytochemistry.  1976.  
Iowa State University Press, Ames IA.  326 pp.

Good luck!

Karen Snetselaar
Asst Professor of Biology
St. Joseph's University
Philadelphia PA
ksnetsel at sju.edu
: --
: Brandi Baros   barosb at alleg.edu
: Allegheny College Bio dept. undergrad

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