wanted: replacement for lactophenol- anilin blue: summary of answers

Yvan Lindekens yvan.lindekens at rug.ac.be
Wed Sep 16 00:04:35 EST 1998


"Lactophenol is used as ... Does anyone know of a substitute for these
staining solutions: I want to get rid of the phenol..."

Summary of answers:
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From: jmhi at cybercom.net (Jeffrey C. May):

"1% Acid fuchsin in lactic acid works fine.  Don't use the phenol!"
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From: boytrytis at peachnet.campus.mci.net (David Slomczynski):

"I haven't found a replacement, yet. The problem is not neccessarily the
phenol (there are certain over the counter medications that use more phenol
- Chloraseptic and other - we injest it). I don't think I would worry so
much about it - You normally use so little that is shouldn't affect
anything.
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From: dpfister at oeb.harvard.edu (Donald Pfister):

"try lactic acid alone"
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From: Helen TURNER <H.C.Turner at greenwich.ac.uk>

I often use toluidine blue in aqeous solution (0.05% w/v or lower).  
This stains hyphal cytoplasm and spores quite nicely, and I've also 
used it to visualise the hyphae of fungal pathogens within (hand or 
cryotome) sectioned leaf and root material.  Leaf material can be a 
bit of a problem, since hyphae stain purple and mesophyll cytplasm 
stains blue.  Looks OK to the eye, but hard to distinguish in 
black-and-white photographs, if you're looking to publish without 
having to pay for inclusion of colour plates.

I have also used aqueous aniline blue for staining fungi with some 
success.  One other thing you might try, if you have access to a UV 
microscope - a number of fungi fluoresce quite nicely under UV 
illumination, especially if you add some dilute fluorescent 
brightener (e.g. from Sigma-Aldrich).  Works a treat for showing 
pathogens growing over leaf surfaces, but not good for picking up 
hyphae inside host tissues.

Hope this helps,
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Thanks to all!

Yvan Lindekens.










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