SEM protocols

Heiko Schoof hsf at mpib-tuebingen.mpg.de
Mon Jul 21 11:14:11 EST 1997


Hello Chris,

There's a very easy protocol using Methanol which I've tried and which
works fine as long as you're not interested in having cell morphology well
preserved but want to have a look at the "big things", I used it for
flowers and it's fine for pictures of whole arabidopsis flowers, but if you
look at single cells they're kind of sunken-in and not as nice and rounded
as they are when I use FAA (Formaldehyde-aceticacid-alcohol). It's
published: c. Neinhuis and H.G. Edelmann, journal of microscopy, 184, 1996,
pp 14-16. It's really simple: just throw the tissue into 100% Methanol,
leave for some minutes, exchange for dry ethanol once or twice, then do
critical point drying and sput. All in a couple of hours from harvest to
microscope...if you want to see what it looks like, look at the flower
picture on our homepage, it's a SEM tinted by Photoshop.
http://www.lmb.uni-muenchen.de/groups/schaeffner/Groups/Laux.html

If you want to, I can send you our "extensive" protocol (takes some days)
using FAA and OsO4, but I've only got it typed down in german. The cellular
morphology is superior, so for SEM of embryos and meristems/flower
primordia we use that.
For pollen SEM, you can contact Paul mailto:egrini at mpib-tuebingen.mpg.de ,
does that, I believe.
Good luck!
Heiko


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