[Cytometry] allophycocyanin (APC) gets very dim

Clintrial clintrial at gmail.com
Fri May 20 11:34:29 EDT 2011


Thanks a lot for the input. The core manager ran 197 beads, and the APC
looks very bright. So, I guess there must be something to do with my
staining.
Zhifei

On Fri, May 20, 2011 at 8:23 AM, Alphen, Floris van
<f.vanalphen at sanquin.nl>wrote:

> 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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