[Cytometry] Colour compensation

Michael Waring mwaring708 at gmail.com
Thu Apr 18 13:39:54 EDT 2013


Actually, I've heard that you might need to compensate the scatters, if you
have something like GFP--the scatter detectors are measuring LIGHT, just
like the other channels, so any emission in their range of detection will
be measured--GFP actually has emission about 20% of maximum at 488, so that
light would "spill over" into the FSC and SSC detectors and make the GFP+
cells seem larger than the GFP negative cells... That said, I dont know of
any instruments that actually allow for this correction?

Mike Waring
Ragon Institute
Massachusetts General Hospital


On Thu, Apr 18, 2013 at 6:11 AM, Vivek Tanavde <vtanavde at hotmail.com> wrote:

> Hello Rabab,Why would you want to do compensation on side scatter? As far
> as I know compensation is necessary for bleed through of a fluorescent dye
> in other PMTs detecting fluorescence. Since the SSC detector does not
> detect fluorescence, it is not necessary to compensate the signal on this
> detector.Regards,
>
>
> Vivek Tanavde
>
> Bioinformatics Institute
> Singapore
>
> From: rababhaider3 at hotmail.com
> To: cytometry at lists.purdue.edu
> Date: Tue, 16 Apr 2013 19:18:33 +0400
> Subject: [Cytometry] Colour compensation
>
> Hi everyone!
> I'm using FC500.
> Is there anyone uses FC500 and knows how to do the automated compensation
> on SS Log scale. All the options from auto setup schedule in the software
> uses FS Lin/ SS Lin.
> Thank you,
> Regards,
> Rabab Al lawati
> Royal Hospital
> Oman
>
>
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