[Cytometry] Colour compensation

Rene.Brons at crp-sante.lu Rene.Brons at crp-sante.lu
Fri Apr 19 10:22:48 EDT 2013


The IDEAS software of the ImageStream allows to compensate on brightfield 
(FSC)

Brons N.H.C. Ing.
CFC (Core Facility Flow Cytometry) / CRP-Sante
CHL, Maternity
sous-sol, porte 7

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From:   Michael Waring <mwaring708 at gmail.com>
To:     "cytometry at lists.purdue.edu" <cytometry at lists.purdue.edu>, 
Date:   04/19/2013 03:07 PM
Subject:        Re: [Cytometry] Colour compensation
Sent by:        cytometry-bounces at lists.purdue.edu



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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