Posted by
Jan Grawe on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Acquiring-a-multiphoton-system-some-questions-tp592485.html
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