[3dem] Chemical fixation before plunge freezing or HPF

[Trepout Sylvain]

Dear Linda,


When working with cells (eukaryote or prokaryote) that need fixation because of safety issues, I usually fix them in 4% PFA.

This protocol never gave broken/damaged cells.


On what type of sample do you want to use chemical fixation ?


Kind regards,

Sylvain

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Sylvain TREPOUT, PhD, HDR
IR2 Inserm
Plateforme de Microscopie Electronique
INSERM US43 / CNRS UMS2016
Institut Curie Centre De Recherche
Centre Universitaire, Bât. 101B-110-111-112
Rue Henri Becquerel
CS 90030
91401 ORSAY Cedex, FRANCE
Phone : +33 1 69 86 30 81
Fax : +33 1 69 07 53 27
________________________________
De : 3dem <3dem-bounces at ncmir.ucsd.edu> de la part de Linda Sandblad <linda.sandblad at umu.se>
Envoyé : lundi 18 janvier 2021 12:18:42
À : 3dem at ncmir.ucsd.edu
Objet : [3dem] Chemical fixation before plunge freezing or HPF

Dear best colleagues!

What chemical fixation protocols works for your projects before plunge freezing (or HPF) or what typs of chemical fixation was not good (destroyed more than it helped) prior to cryo-EM sample preparation?

Im interested in your experiences, also happy if you have adressed this question in a article and would like to share the link with me.

For cryo-EM we prefer to work as close to native as possible, avoiding chemical fixations, but sometimes a fixation is needed to test or move on with tricky projects. For examples, when bacteria cultures are tricky and samples need to be stored, or when bio safety restricts work in the EM labs (lab specific and different).

Best regards, Linda

Linda Sandblad, researcher and facility director
Umeå Core Facility for Electron Microscopy, SciLifeLab Cryo-EM, NMI
Department of Chemistry, Umeå University, Sweden
Tel: +46 709324936

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