[3dem] EtOH vs Ac.
reinhard rachel
reinhard.rachel at biologie.uni-regensburg.de
Sun Feb 21 09:29:14 PST 2010
we have been using Ethanol as the main solvent in freeze-substitution fixation for many years, when processing prokaryotic cells, and are happy. In direct comparison with Acetone+OsO4, we get similar but not identical, in some cases even superior results; fine structures in the cytoplasm become visible, which are not (or rarely) seen in any Acetone/Os-substituted sample.
We use: Acetone + OsO4, now mostly with 0.1 to 0.25% UAc, plus up to 5% Water
Ethanol + 0.5% UAc + 0.5% GA (+1% FA) + 5% water.
While we have not tested yet the Acetone / Os samples for their antigenicity, we are successful in post-embedding labelling (resin: Epon) following Ethanol +U + FA/GA+water substitution, using a variety of antibodies.
Considerations? theory is the one side (look for papers by Bruno Humbel & Heinz Schwarz, K. McDonald and many others) - experience is the other side. If it works ... be happy.
Hope this helps ... Reinhard
--
PD Dr. Reinhard Rachel
Universitaet Regensburg
Centre for EM - NWF III -
@Institute for Anatomy
Universitaetsstr. 31
D-93053 Regensburg - Germany
tel +49 941 943 2837, 1720
fax +49 941 943 2868
mail reinhard.rachel at biologie.uni-regensburg.de
office: VKL 3.1.29
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