Re: Yield problems

If your cells are big and fragile as I suspect they are they may explode when
they leave the nozzle. Try using a 100u nozzle and lowering your pressure to
21psi or something.
Simon

Quoting "Christopher A. Worth" <caw@bcc.louisville.edu>:

>
> what type of tubes are you catching your cells in?
>
> Polystyrene by chance?
>
> I've seen the catch tubes get a charge built up on them and actually deflect
> the droplet
> away.
>
> Now this was on an Elite, with much lower droplet velocity.
>
> I figure if something isn't making sense, you have to check everything.  Are
> you getting
> a droplet forming on
> the rim of the catch tube?  That's the telltale sign.
>
>
>
> On Tue, 29 Apr 2008 14:32:29 +0200, Simona Ronzoni wrote:
>
> >Dear Flowers
>
> >I have a problem with FacsAria
> >When I sort stem cells from KASUMI cells linee (Acute
> >MyeloId Leukaemia), I lost the cells.
> >My guess is that the cells are being lost due to stickiness or cell
> >death.
> >Looking at FSC vs. SSC and gating on the viable cells post sort only
> >10% of the events fall in what I would consider debris or dead cells. The
> >cells are spin down	and then counted. There are essentially no dead cells
> >visible
> >in these counts.
> >Last time I sorted 3.200.000 cells and I have count only 11.000!! It's
> >impossible!! I don't have the same  problem with other cells type and I use
> >always the same parameters ( PSI, Drop delay and Frequency)
> >Can you help me?
> >Any insights will be greatly appreciated.
>
> >Ciao
> >Simona
>
> >--
> >Simona Ronzoni
> >Imaging Centre
> >Consortium for Genomic Technologies
> >c/o IFOM-IEO Campus for Oncogenomics
> >via Adamello 16
> >20139 Milan
> >Italy
> >Phone: ++39-0294375087/0294375038
> >email: simona.ronzoni@ifom-ieo-campus.it
> >http://www.ifom-ieo-campus.it/RESEARCH/Imaging/
> >http://www.ifom-ieo-campus.it
>
>
>
>
>
>
>


-- 
Simon Monard
Cytometry Lab Manager
Rua do Campo Alegre, 823,
4150-180 Porto - Portugal
Tel +351 226 074 900 ext 3102 . Fax +351 226 099 157
Cellphone # 919 036 680

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