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Hello, Shiv & All -
We have 2 Zeiss NLO 2-photon sytems, both in beige-walled rooms. The NDD
detectors are much more sensitive to stray light. As others have mentioned,
the monitors are the biggest problem. A curtain works well or turning the
brightness/contrast all the way down can also help. But even so, we either
cover the front of the microscope on the upright system or put a "cover" or
"blocker" over the specimen on the inverted - a dark piece of hard foam is
the usual choice. Aside from that, we have had to cover all LEDs in the room
and on the microscope itself. Black electrical tape works pretty well or
even lab tape. We have a curtain between the microscope and the monitors
and doorway on one of our rigs. When the curtain is closed, we can even open
the door to the room and not affect the imaging. Try not to cover the
scanhead on the microscope b/c that should not be allowed to heat up too
much. I wouldn't worry about the color of the walls. If you've gotten rid
of the light sources, it won't matter that light walls reflect more than
dark.
Good luck!
-Holly
CRL Molecular Imaging Center
UC Berkeley
http://imaging.berkeley.edu
-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On
Behalf Of Mcnamara, George
Sent: Friday, May 20, 2005 8:36 AM
To: [log in to unmask]
Subject: Re: Zeiss LSM 510 NLO 2P Room Color and NDD Detectors
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As Karl Garsha noted, room lighting - i.e. the LSM's computer monitors, are
more of a problem than the wall colors.
From the recent UCHC course I attended, the non-descanned ("direct", though
in fact they are back by the arc lamp, not under the objective lens) are
about 4x more sensitive than the PMT's in the confocal scanhead, which are
in turn about 4-5x more sensitive than the META detector. With the
non-descanned detectors it is easy to see the contributions to background
intensity of the room lights, light from the doorway, light from any desk
lamps, and even light from the LEDs and switches. Although inconvenient,
when I use the non-descanned detectors here I usually turn off the arc lamp
to (1) eliminate stray light from it reaching the non-descanned, and (2)
eliminate "user error" of blasting the detectors with direct arc lamp
illumination (I'm still getting the hang of the shutter sliders on the
LSMs). For the latter, I've been told that PMT's will recover in ~24 hours -
a claim I hope never to test.
George
-----Original Message-----
From: Sivaguru, Mayandi [mailto:[log in to unmask]]
Sent: Thursday, May 19, 2005 3:19 PM
To: [log in to unmask]
Subject: Zeiss LSM 510 NLO 2P Room Color and NDD Detectors
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We are going to acquire a Zeiss LSM 510 META NLO 2P system with NDD
detectors. I have seen places where it was housed in rooms with either white
or black painted walls. We have also received comments that NDD detectors
are sensitive to stray or reflected light. Despite we do not have any hands
on data as of now using NDD detectors, we would like to know, how far the
room wall color is important while using the NDD detectors.
Thanking you in advance and I appreciate all your insights on this.
Shiv
Mayandi Sivaguru, PhD
Associate Director
Molecular Cytology Core
Biomolecular Research Facilities
120 Life Sciences Center
University of Missouri
Columbia, MO 65211
Voice: 573-882-4895
Fax: 573-884-9676
[log in to unmask] <mailto:[log in to unmask]>
www.biotech.missouri.edu/mcc <http://www.biotech.missouri.edu/mcc>
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