[Cytometry] allophycocyanin (APC) gets very dim

Alphen, Floris van f.vanalphen at sanquin.nl
Fri May 20 11:23:00 EDT 2011


Hello Zhifei,

I would check your delay and/or alignment of the red laser.
Good luck,


Floris

Floris van Alphen
Dept. of Molecular Cell Biology/
Central facility for cell analysis
Sanquin Bloodsupply, research division
room U264A
Plesmanlaan 125, 1066CX Amsterdam
the Netherlands
+31(0)20-5123250
F.vanalphen at sanquin.nl



-----Original Message-----
From: cytometry-bounces at lists.purdue.edu
[mailto:cytometry-bounces at lists.purdue.edu] On Behalf Of Clintrial
Sent: vrijdag 20 mei 2011 9:20
To: cytometry at lists.purdue.edu
Subject: [Cytometry] allophycocyanin (APC) gets very dim


Dear Flowers,

I recently ran into a dim APC problem. Our lab have been using LSR II,
and could get very bright signal in APC compensation control using BD
CompBeads (I mixed neg and pos beads in one tube when labeling
APC-conjugated Rat anti-mouse IgG 2b), and good separation of my cell
population of interest until last week. All of a sudden, for some
reason, I could not get good separation of APC neg and pos signal any
more in both compensation control and cell labeling. I checked the LSR
II voltage, identical; my labeling protocol, solution, are all the same;
I also ordered the new antibody, still I can never get bright APC signal
any more.

Does anyone has similar experience and please could you share your
thoughts as to what could be the possible reason?

Zhifei
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