[Cytometry] UV 20 or 60 mW
flowmail at verizon.net
Tue Apr 16 13:18:14 EDT 2013
If you mean 561 for PE and tandems, at a certain point you will saturate the fluor.
Different case for UV excited dyes. I'm not sure it will help SP all that much. My experience is alignment is the critical issue.
For cell cycle, more power will give you smaller cv in most cases (if your cells will actually have small cv). But if you are sorting cell cycle populations, higher UV may have severe DNA damage artifacts. Consider that for your downstream uses.
On Apr 16, 2013, at 8:29 AM, "Schmitt, Steffen" <steffen.schmitt at Dkfz-Heidelberg.de> wrote:
> Dear colleagues,
> I would like to hear or better read your opinion/ experience regarding 20 mW
> UV-laser versus 60 mW UV-Laser.
> We are working with an upgraded AriaIIu and our 20 mW UV laser died.
> The question is, if we replace it with another 20 mW or if we spent the
> money for an 60 mW UV-Laser.
> Can you get a better resolution (e.g on Hoechst or DAPI cell cycle or Side
> population) with more laser power?
> I remember a threat some time ago, that more power with a 561 nm laser not
> necessarily gain more resolution or better sensitivity.
> Thanks for you input.
> PS: Our Ca-Influx assay is principal working, but we now have to figure out
> the right experimental activation conditions. Thanks for your help regarding
> this threat.
> Dr. Steffen Schmitt
> Head of the Flow Cytometry Unit
> German Cancer Research Centre
> (Deutsches Krebsforschungszentrum in der Helmholtz-Gemeinschaft,
> Stiftung des öffentlichen Rechts)
> Imaging and Cytometry Core Facility
> Cost center: W220
> Im Neuenheimer Feld 280
> D-69120 Heidelberg
> Phone: +49-(0)6221-42-1261 (office)
> +49-(0)6221-42-3643 (lab)
> Fax: +49-(0)6221-42-3755
> E-Mail: steffen.schmitt at dkfz-heidelberg.de
> Web: www.dkfz.de/en/facs/index.html
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