Clearing agents

Frederique Guinel f fguinel at MACH1.WLU.CA
Thu Nov 6 15:55:27 EST 1997


 Hi Christine,

In my plant anatomy class, I use the following recipe.

	Place tissue into methanol and heat gently to just below boiling 
point.  Hold for several minutes until chlorophyll or other pigments are 
extracted.  If the solution becomes discoloured, replace it with fresh 
ethanol.

	Place decolourized tissues into 70% lactic acid.  Heat to just 
below the boiling point.  Thin tissues will be adequately cleared in less 
than 30 min.  Thin tissues can also be left several days in lactic acid 
at room temperature.
	Thick tissues will require initial heating in lactic acid 
followed by further soaking for about 2 weeks.  Such difficult material 
can be rapidly cleared by autoclaving in lactic acid but this method has 
the disadvantage of browning the tissues.

	Mount unstained cleared tissues in lactic acid; this is sufficient for 
doing optical sectioning with the microscope.  The tissue can 
also be stained depending on what you want to see.  I have some recipes of 
stain for viewing xylem and phloem.

Hopefully this will help.

Frederique C. Guinel
Department of Biology
Wilfrid Laurier University 
Waterloo, Ontario N2L 3C5.
Canada
 

On 6 Nov 1997 CMAXWELL at trentu.ca wrote:

> Dear Plant edders
> 
> I have been looking for a substance which will clear plant tissues 
> effectively.  I remember years ago (when I was a student) that we used 
> Chloral Hydrate to clear leaves. I cannot remember the exact technique, but 
> I also remember that the chemical was declared a dangerous substance. Are 
> their any alternatives? Does anyone know of any other techniques?
> Thanks
> Christine
> 



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