We are also dealing with some of the issues that Alex Bell
asked about and Katheryn Resing commented on regarding
2-D imaging and image analysis. Our gels are now
fairly reproducible and give very nice patterns with silver
staining. We have also begun to identify some of the
stronger spots with MALDI peptide mapping, and hope
to increase our success rate with low-flow LC and
ESI on the LCQ.
We will be evaluating the SYPRO stains at a later date, but
the reality of today is silver and coomassie stains.
So, now it's time to image the gels and pull out differences
in spot patterns.
There appear to be 4 major imagers that can handle _both_
colorimetric stains (silver, coomassie) and the fluorescent
protein stains (SYPRO) - the Bio-Rad Fluor-S or Fluor-S Max,
the Hitachi CDBIO 8C or 16C, the Genomic Solutions Investigator Proteomic
Analyzer Camera-based system, and the Alpha Innotec
Fluor-Chem. The Pharmacia system appears to be undergoing
some revisions/improvements and I couldn't find enough
info about it for comparison purposes.
We are not doing any rad work, so the phosphor imagers,
while nice, are not going to be useful for us.
Anyone have any experience with these imagers
that they would be willing to share? We have another
Fluor-S in the building, and the folks who use it for
lane-based DNA gels really like it.
Katheryn brought up a good point about the 8 bit limit of
some of the software programs. But if, as she suggests,
there may be major innovation in 2-D analysis software
soon, that limit may be overcome and a higher bit image
may become necessary. So one question is whether it
is a good hedge bet to go for a 16 bit camera system now
(vs. a 12 bit camera system) to avoid running into a resolution
limit in the future.
We are also looking at image analysis programs, and
one that wasn't mentioned previously is the new version
of Image Master 2D Elite from Pharmacia. It looks like
it would fit somewhere between PDQuest and Melanie
and I am trying to get my hands on a demo.
Anyone tried this package?
Any info is appreciated.
-Dave
************************************
David Dogruel
Los Alamos National Laboratory
Bioscience Division
MS M888
Los Alamos, NM 87545
Ph: 505-665-9071/8034
FAX: 505-665-3024
dogruel@lanl.gov
***********************************
This archive was generated by hypermail 2b29 : Mon Jun 12 2000 - 13:09:42 EDT