Detecting Hoechst 33342 emission on an Epics 753
Antony Bakke (bakkea@ccmail.ohsu.edu)
Fri, 26 May 95 13:45:02 PST
I'm interested in looking at Hoechst 33342 emission along with 3
color immunofluorescence on a dual laser Epics 753 using a blue
line from one laser and UV from the second. Currently I have
the UV coming from the front laser and the blue from the back.
This is the reverse of the usual configuration, but I'm told
that routing the UV through two mirrors from the back laser
would significantly reduce the UV power.
Two questions:
1. Is there any problem with my laser setup?
2. What are the optimal dichroic mirrors and filters for
Hoechst 33342? The Epics 753 did not come with a standard setup
for Hoechst. I have tried all the combinations that I can think
of from the Epics standard filter set, but get only scattered
light coming through the PMT from the blue laser. The only
laser blocking filter in the standard set for the 753 blocks all
emission from 430-520, which is exactly where Hoechst emits.
I'd greatly appreciate any help.
Tony Bakke
e-mail: bakkea@ccmail.ohsu.edu
FAX: (503) 494-5799
phone (503) 494-4687
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