Hi All
 
I am a PhD student in the zoology department at Wits University, South
Africa. I am looking into using the confocal microscope which is a
recent acquisition by the university. I would appreciate any
advice/help that you may have at your fingertips.
 
Let me briefly explain what part of my research entails. I am looking
at the abdominal tergal glands in honeybees - These glands are
unicellular, subepidermal glands. I have done TEM on glands present in
queens and now I need to describe these glands in workers. I have
processed the workers 3-4 months ago and the tissue is embedded in
epon araldite. I realize that fixed tissue is the least ideal for a
number of reasons. I do understand that for best results I should
start from scratch and not fix and embed the tissue. Unfortunately,
that is not so simple because obtaining the material proved somewhat
problematic and having reached this point I really would like to
prevent having to throw away my material and starting from scratch.
 
I am hoping to section these blocks and then view these
sections under the confocal. I would like to describe the fine
structure (ultrastructure not necessay) of the glands and also do some
volumetric analysis. However, the problem facing me is which
post-fixative/post-embedding stain is suitable and highly reflective.
I was wondering whether anyone has been faced with a similar problem
and has found a suitable stain which penetrates resin. Also, maybe
there is somebody combining TEM with confocal work and has already
crossed the bridge concerning the stain. To date I have not found a
fluorescent stain that will penetrate araldite. It would appear that I
should be looking at reflective dyes rather than a fluorescent stain?
I have been given some suggestions but I would appreciate any other
suggestions before tackling my problem. (Wavelength of confocal falls
in the range 300-500nm)
 
Any/all advice will be much appreciated.
 
Thanks a mil
Theresa
email address: [log in to unmask]