[3dem] negative staining artifact

[Matthias Wolf]

Hi Linda,

Looks like your sample is packed with protein – it’s negative stain, protein is white – try dropping the concentration 10 to 50 times. Also, if those spherical objects are lipid or mixed lipid-detergent vesicles, they may not like the UA – try a different stain at neutral pH like PTA.

   Matthias

From: 3dem [mailto:3dem-bounces at ncmir.ucsd.edu] On Behalf Of Linda Sandblad
Sent: Monday, September 25, 2017 6:25 AM
To: 3dem at ncmir.ucsd.edu
Subject: [3dem] negative staining artifact

Hi all experts!!
I know, this do not look nice, but I would like to hear your opinion about this artifact. Tiny dark spots covering both sample and background, do you know what it is, were it comes from and how to avoid it? We have this bad background time to time from different labs…. It is a “normal” negative staining protocol: copper grids, formvar, carbon, glow discharge, UA.
I’m happy for your suggestions, Best Linda
[cid:image002.jpg at 01D335E0.C463FD30]

Linda Sandblad
……………………………………………….
Department of Molecular Biology
Umeå University
+46 709324936

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