Real-time signaling on an LSR II or CyAn

McArthur, Cliff Cliff.McArthur at ucsf.edu
Wed Jan 24 16:08:45 EST 2007


Hello Flowers-All,
 
My seriously sincere apologies if this is a duplicate message.	I
thought I'd sent this inquiry out last week, but having heard absolutely
zero back, I think I made an erroneous keystroke somewhere.  I'd have
expected at least a dozen out-of-office replies... I've searched the
archives, too, and found nothing.
 
We have an interest in doing some real-time signaling by flow and are
trying to imagine the contraption of sample tubes we might need to build
which could help us accomplish this.  We'd like to track Ca2+ flux, of
course, as it's affected by both ionophores and inhibitors, and the
experiment, in theory, is simple: baseline measurement on sample -
add/inject ionophore and observe flux - add/inject inhibitor - observe
effect - attempt to wash out/off inhibitor and observe...
 
Follow?  Of course you do :-)
 
We're imagining a sort of "Y" shaped contraption of assembled 12x75 (5
mL) tubes (we don't have an HTS) which could allow us to add these
reagents sequentially and wash out while adding no or known volumes of
PBS.  Question is.... Anyone else imagine such a contraption or assemble
some other one to attempt real-time signaling by flow??
 
Huge Thanks for any input!
 
Cliff J McArthur, Specialist
Flow Cytometry and Cell Sorting
Sandler Sorting Facility
University of California, San Francisco
513 Parnassus Ave. S1067, Box 0795
San Francisco CA 94143-0795
 
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