[Cytometry] PI intensity moves during analysis

Alexandre Bote Tronchoni abote at ir.vhebron.net
Wed May 11 03:21:34 EDT 2011


Hi all,

We have a Aria1 and we noticed that the injection sample tub is stained with
PI....Have you noticed that? Did you wacht the injection sample tub before
and after pass a sample? If the amount of PI is very high the injection
sample tub is stained and then the signal is increasing over the time cause
the sample is being stained trogh the capilary. You should mark the cells
with less PI. I don't know if that is your problem or is related to the
pressure changes but This could be one of the reason of the change in PI
over time...Is always increasing the value?

I'll hope it could help you (or anyone else). For cleaning the injection tub
just pass FacsClean and FacsRinse or any other cleaning solution for a long
time and try with less amount of dye.

Good luck
--------------------------------------------

**


A*lexandre Bote Tronchoni*

*Plataforma de Citòmica
UCTS
*Institut de Recerca
Hospital Universitari Vall d'Hebron
Tel: 93 489 41 79

Fax: 93 489 41 85
abote at ir.vhebron.net
http://www.ir.vhebron.net



2011/5/11 Rossi Ralph <ralph.rossi at petermac.org>

>  Hello All,
>              I have seen this as well, to greater and lesser degrees
> and SUSPECT it's some sort of equilibrium process
>                  between the PI solution the cells are in and the
> tubing/flow cell ?
>
> ralph
>
>
> Regards
> Ralph Rossi
> Flow Facility Manager
> Peter MacCallum Cancer Institute
> Melbourne , Australia
> Ph 96563747 , 96561955
> Email : ralph.rossi at petermac.org
>
> -----Original Message-----
> From: cytometry-bounces at lists.purdue.edu
> [mailto:cytometry-bounces at lists.purdue.edu] On Behalf Of Nebe-Von-Caron,
> G
> Sent: Wednesday, 11 May 2011 4:42 PM
> To: Elisabeth Freyer; Cytometry at lists.purdue.edu
> Subject: Re: [Cytometry] PI intensity moves during analysis
>
> If you reload the same sample a second time do you get the same change
> with time?
>
>
> Gerhard Nebe-von-Caron
> Research Scientist and Biomedical Engineer
>
> -----Original Message-----
> From: cytometry-bounces at lists.purdue.edu
> [mailto:cytometry-bounces at lists.purdue.edu] On Behalf Of Elisabeth
> Freyer
> Sent: 10 May 2011 14:35
> To: Cytometry at lists.purdue.edu
> Subject: [Cytometry] PI intensity moves during analysis
>
> Hello everybody,
>
> I am observing some strange shifts of the PI intensity when i do cell
> cycle analysis. I'm not talking about the shifts between samples, my
> whole histogram of the same sample actually moves up in intensity within
> the first minute of running it on the machine. After a minute it stays
> about the same.
>
> I use an Aria2, measure PI in 488-685/35BP It ONLY happens when I use
> protocols that require treating the cells with Pepsin or Trypsin (e.g.
> Vindelov) prior to  PI staining. no movement w/ EtOH fixation and
> staining in PI/RNAse solution only.
> The samples are sitting on ice until they get loaded onto the machine.
> I thought it might be temperature related, but also with a cooled sample
> chamber it still moves.
>
> To record I now usually wait for a minute for the PI intenisty to
> stabilize and record then. It gets a bit difficult when I have samples
> with very small cell numbers :(
>
> I'd greatly appreciate if somebody could enlighten me where that comes
> from and if there is anything I can do to stop this.
>
> Cheers
> Lizzie
> :)
>
> --
> Elisabeth Freyer
> Flow Cytometry Facility
> MRC Human Genetics Unit
> Western General Hospital
> Crewe Road
> Edinburgh
> EH4 2XU
>
> Tel: 0131 332 2471
> email: Elisabeth.Freyer at hgu.mrc.ac.uk
>
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