Re: FCS of proteins in solution

Posted by Csucs Gabor on
URL: http://confocal-microscopy-list.275.s1.nabble.com/FCS-of-proteins-in-solution-tp590361p590362.html

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Dear Els,

Proteins do adsorb everywhere, pretty much independent on the buffer you
are using (of course the amount can be varying). On way to avoid this
that you start your experiment with a high concentration of proteins -
let the saturate the surface and start you FCS measurements afterwards.
Actually if you use a large enough cuvette/sample volume and you measure
the FCS signal as far from the glass as it is possible - the this should
solve your problems. Another way to deal with your problem avoid this
(as someone mentioned it before) is the passivation of the surface. This
can be achieved either by incubating your glass slide in serum albumin -
this is the simplest way (although not particularly stable), but you can
use other , dedicated materials like Pluronics or PLL-g-PEG (we have
published couple of papers with this latter). But there are of course
many other materials which one could use. Surface passivation is a
rather"standard" biomedical/biotechnological problem so there are many
papers existing on the subject.

Cheers    Gabor

--
Gabor Csucs
Light Microscopy Centre, ETH Zurich
Schafmattstrasse 18, HPM F16
CH-8093, Zurich, Switzerland

Web: www.lmc.ethz.ch
Phone: [phone]
Fax: [phone]
e-mail: [hidden email]