SP Cells

WEHI Cytometry facs_copy at wehi.EDU.AU
Mon Nov 28 18:44:01 EST 2005


Leonie,

I think your filters should work fine.	On our DiVa we use around 450 
volts for the blue and up to 800 volts for the red (give or take 50V) 
in order to put the major viable population (G0/1) at about mid-scale 
in both UV1 and UV2 directions, both on a linear scale, zero 
compensation for both, with UV laser power of 50mW.  In our experience, 
getting the staining right is much more difficult than getting the 
instrument setup right.  Timing is critical.

Frank Battye.


> I am having a bit of trouble setting up the Voltages and Compensation 
> for
> SP Cells.
> I am acquiring the cells on a BD FACSVantage with DiVa option in 
> Digital Mode.
> How critical are the filters,I have a Hoechst 33342 450/20 for the 
> blue and
> a 660/20 for the red but I only have a 525DCSP for the splitter. Are 
> these
> okay or not specific enough?
>
> I also have a problem in the two papers I am referring too. One paper
> states that the detectors should be in Linear mode while a second 
> paper is
> in Log. Up till now I have been running in Log so which is correct and 
> I
> would be very interested to hear anyones experiences.
>
     |	  |  << Frank Battye PhD
      \__/ <<<< The Walter & Eliza Hall Institute
------!!<<<<<< 1G Royal Parade, Parkville
      /!!\ <<<< Victoria 3050, Australia
     o !! \  << ph: 61_3_9345 2541, fax: 61_3_9347 0852

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