*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****
Are you sure there is a 514nm DPSS? I think that wavelength would have
to be an OPS and I don't know anyone who makes one. Coherent do make a
505nm OPS which is intended to stand in for both the 488 & 514 lines of
an Argon laser, and I rather suspect that this could be exactly what you
need. But I have no experience of it. (I do have a Coherent 488nm
OPS).
Guy
Optical Imaging Techniques in Cell Biology
by Guy Cox CRC Press / Taylor & Francis
http://www.guycox.com/optical.htm
______________________________________________
Associate Professor Guy Cox, MA, DPhil(Oxon)
Australian Centre for Microscopy & Microanalysis,
Madsen Building F09, University of Sydney, NSW 2006
Phone +61 2 9351 3176 Fax +61 2 9351 7682
Mobile 0413 281 861
______________________________________________
http://www.guycox.net
-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]]
On Behalf Of Tim Feinstein
Sent: Thursday, 10 November 2011 7:34 AM
To: [log in to unmask]
Subject: FRET in the time of DPSS
*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****
Hello all,
We want to spec a four-laser launch for a new live cell system that will
handle both CFP/YFP FRET and red/green imaging. However, I am sad to
see that gas lasers are no longer speccable and so the freebie 514 laser
line is gone. We would therefore have to spec a 514 DPSS and forego the
far-red line.
I was wondering whether there is a way to do more (or at least the same)
with less. 488 nm excites YFP well enough, so in theory I could image
CFP/YFP using a scan head dichroic with cutouts for 442 and 488 nm laser
lines. In my experience 442 nm laser excitation (via TIRF) causes
negligible YFP excitation and 488 nm does not excite CFP, so it is
possible that I could gain speed by passing everything through a single
broad bandpass filter (e.g., 455-550 nm) and alternate excitations.
Assuming that cross-talk is not a problem, the most significant cost
would be that I lose a decent chunk of CFP emission to the scan head
dichroic, but in return I gain a 641 nm laser.
Has anyone tried this? Any feedback on or off-list would be much
appreciated.
Thanks and all the best,
TF
Timothy Feinstein, PhD
Postdoctoral Fellow
Laboratory for GPCR Biology
Dept. of Pharmacology & Chemical Biology
University of Pittsburgh, School of Medicine
BST W1301, 200 Lothrop St.
Pittsburgh, PA 15261
-----
No virus found in this message.
Checked by AVG - www.avg.com
Version: 10.0.1411 / Virus Database: 2092/4005 - Release Date: 11/08/11
|
