[3dem] HPF

[Reinhard Rachel]

dear Xavier, 

-- 
Prof. Dr. Reinhard Rachel
University of Regensburg
Centre for EM / Anatomy
Faculty of Biology & Preclin. Med.
Universitaetsstrasse 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 
 
next microscopy conferences: 
http://www.emc2012.org.uk/
EMC 2012 in Manchester, UK
http://www.mc2013.de/
MC2013 in Regensburg, Germany


>>> 13.09.2012 um 17:35:
> I am considering High Pressure Freezing a specimen that is in a serum 
> with 2.5% wt/vol Methyl-cellulose. I am not sure if the MC will will not 
> limit the vitrification efficiency or induce ice crystal formation.
> And later for the freeze substitution if it will give me troubles with 
> Lowicryl and/or EPON.

it is worth thinking about experiments in advance, yes... but in this case it makes sense (in my opinion) to try it out! In particular, as there are more and other variables in this experiment which we don't know yet: what is "the specimen"? what kind of HPF machine are you using? which carriers? (depth, diameter, and so on). and what is the composition of the serum? often, these contain a lot of substances in a fairly high conc which may interfere with the experiment to a degree which is unpredictable. I would guess that MC reduces the risk of ice crystal formation. But, who knows. FS: which solvent would you favor to use? many cocktails are in use ... 
Go ahead and let us know ... 
kind regards - Reinhard