4/3/97


Dear Microscopists:

Can anyone please comment on the storage properties of common EM/LM fixatives.

We have some older 1% glut/4% formaldehyde in PO4 buffer. Is there some rule

of thumb that people are using, should we analyze before use, are there some

references we can access?

Thank you in advance for your comments.

Regards,

Ken Baker

bakerk@aa.wl.com


The aldehydes oxidise to acids - formic or glutaric. The reaction is less

at lower pH so is accelerated when you buffer the solution to pH7. Our rule

is to buffer the fixative just before we use it and discard any buffered

fixative older than a week.

Mel Dickson

M.Dickson@unsw.edu.au


The question was how long fixatives can be stored in an TEM lab.

After exhaustive investigation some years ago, we arrived at the

following answer:

The highest grade glutaraldehyde or paraformaldehyde (distilled and

stored in glass vials under inert gas) will deterioate to approximately

one half their strengths in 3 weeks assuming they are in a buffer,

approximately at pH 7.4, and under continuous refrigeration. Therefore we

never keep fixatives for more than one week. We make them up the day of,

or the day before we use them.

Consider how valuable your sample is and how perfect it is expected to

look. A sample which has undergone autolysis will not benefit by

ultra-fresh fixation fluids, but living cells in culture will.

Oxygen and heat are deleterious to aldehydes. Glutaraldehyde in a mostly

empty bottle will not last very long, while glut in a glass vial sealed

under inert gas will last many years.

Hope this helps.

HILDEGARD CROWLEY

hcrowley@odin.cair.du.edu


We sometimes store small aliquots of fixative at minus 20 C. I'm not sure

this is effective but it seems like a reasonable thing to do. Seems as

though I came upon this in one of the early editions of Hyats general EM

techniques book.

*******************************************************

G.W. Erdos, Ph.D. Phone: 352-392-1295

Scientific Director,

ICBR Electron Microscopy Core Lab

218 Carr Hall Fax: 352-846-0251

University of Florida E-mail: gwe@biotech.ufl.edu

Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/


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