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Armstrong, Brian on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Acquiring-a-multiphoton-system-some-questions-tp592485p592486.html
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http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocalHi Jan, 1. I am having Zeiss and Coherent come out together to hopefully gain some insight into this question. Certainly the AOM will cause GVD, how much is the real question. 2. I agree with George that experts suggest that pre-chirping isn't really worth the bother. One question that I have never received a satisfactory answer to is how you adjust the pre-chirp every time you change lamda, the interprism distance would need to be altered to account for the differing chirp from the differing lambda. Both Spectra (MaiTai Deep See) and Coherent, offer pre-chirp options for their lasers.
3. Carolina makes prepared slides that are very thick. I have the pig embryo.
http://www.carolina.com4. That is an excellent question. It troubles me. I hear that Prairie Tech
http://www.prairie-technologies.com/ "hand picks" their Hamamatsu PMTs. I think they are identical to the ones Zeiss used in their LSM510 NDD systems. I don't know if this would account for the difference, but the Prairie system is much more sensitive than the 510 NDD system. Zeiss will have new and improved (custom made?) PMTs on their LSM 710s. We will have to get the specs as they slowly leak out from Zeiss.
Brian D Armstrong PhD
Light Microscopy Core Manager
Beckman Research Institute
City of Hope
1450 E Duarte Rd
Duarte, CA 91010
626-359-8111 x62872
http://www.cityofhope.org/SharedResources/LightMicroscopy-----Original Message-----
From: Confocal Microscopy List [mailto:
[hidden email]] On Behalf Of Jan Grawé
Sent: Sunday, February 24, 2008 1:28 PM
To:
[hidden email]
Subject: Acquiring a multiphoton system-some questions
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocalDear all,
we are in the process of acquiring a multiphoton
system intended mainly for intravital imaging,
and I have a couple of questions regarding performance:
1. If I understand it correctly (correct me
otherwise!), intensity modulation using an AOM
is a major source of chirp. At least one
manufacturer offers an alternative without AOM,
which is said to improve imaging depth. With the
advent of pre-chirping, is this a relevant issue?
We would of course like the utility and bleaching
capabilities afforded by an AOM, but perhaps not
if it significantly reduces the imaging depth.
2. Connected to the above, to those of you that
have experience with pre-chirping. How much could
we expect this to improve our imaging depth? Is
the improvement greater if we choose a laser with
shorter pulses ~80fs as compared to 140fs?
2. Since no manufacturer will give any hard
numbers with regard to the depth reach of their
systems, we would like to be able to test this
for ourselves. Do you know of a test sample that
can be made several hundred um thick, that is or
mimics some type of tissue, that has specific
fluorescent structures at various depths, and
that preferably is stable at least over some
weeks. Are there commercially available MP test samples?
A tall order maybe, but I´m hoping..
4. What components of the system do you feel are
the most critical for the overall performance. Is
it the laser pulse shape, NDD performance, obcectives, other??
All responses are appreciated!
Best regards,
Jan Grawé
Jan Grawé
Cell Analysis Core Facility
Rudbecklaboratoriet/C5
Dag hammarskjölds väg 20
SE-75185 Uppsala
SWEDEN
Phone: +(0)18-4714657
Cell: +(0)70-2577874
[hidden email]
www.rudbeck.uu.se/cellanalys
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