HeCd laser
Michal Bochenek (mbochen@izoo.krakow.pl)
Tue, 31 May 1994 11:29:54 +0200 (MESZ)
Hello,
I encountered some problems with UV laser (HeCd 325nm, Omnichrome, at Epics
Elite). I cannot receive the correct signal from cells under UV light.
I performed or checked:
1. the light reaches (through beam shaper, fluorescence lenses) the
PMTs;
2. mirror alignment (with targets, et.) - completed;
3. flow cell alignment - completed;
4. beam shaping lens (cylindrical) - 2 types: lengths 48mm & 68mm -
used
5. filters for FS detector - 1.0ND or 325nm bandpass - used;
6. obscuration bars - standard vertical or selfmade horizontal.
The horizontal one, because I heared there is horiz. polarization
plane of this UV light. Several different positions and widths
were tried. Several, but perhaps not all?
After running DNACheck or cells stained with Hoechst 33342 I received
peculiar signal. The increase of FS voltage makes higher data rate (at PMTs
as well) but don't move the particles' "cloud" along the FS axis, though I
can to do this with side scatter signal.
Furthermore, it's funny (but not for me) the signal is the same in presence
or absence cells in the stream.
Cytometer works quite well with "standard" argon & HeNe lasers.
What is my mistake?
I can ask the Service, certainly, but I hope this list is the fastest way to
solve my problem, and I prefer to do it myself.
Could anyone help me?
Mike
Michal Bochenek
NRIAP, Dept.of Anim.Repr.
32-083 Balice/Krakow, Poland
fax: 48 012 110294
CD ROM Vol 2 was produced by staff at the Purdue University Cytometry Laboratories
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If you have any comments please direct them to Dr. J. Paul Robinson, Professor & Director, PUCL,
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EMAIL robinson@flowcyt.cyto.purdue.edu
