Posted by
Lemasters, John J. on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Live-cell-imaging-with-multiphoton-confocal-tp6297623p6300348.html
*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy*****
Hi David and Mark,
Good points. By specialized laser and optics, I meant a UV argon laser and UV quartz optics. I had such a beast once to image NADH, but multiphoton is much easier and more versatile.
Regarding multiphoton photodamage, David Piston has a paper showing that 2 photon imaging is more damaging than 1 photon imaging for cells in culture and that the multiphoton photodamage has a 3 photon power profile. Hence most folks will tell you to use conventional 1 photon imaging for cells in culture -- but you'll still need 2-photon microscopy to image things like NADH and NADPH unless, of course, you have a UV laser and UV optics.
John
--
John J. Lemasters, MD, PhD
Professor and GlaxoSmithKline Distinguished Endowed Chair
Director, Center for Cell Death, Injury and Regeneration
Departments of Pharmaceutical & Biomedical Sciences and Biochemistry & Molecular Biology
Medical University of South Carolina
QF213 Quadrangle Building
280 Calhoun Street, MSC 140
Charleston, SC 29425
Office: 843-792-2153
Lab: 843-792-3530
Fax: 843-792-8436
Email:
[hidden email]
http://academicdepartments.musc.edu/ccdir-----Original Message-----
From: Confocal Microscopy List [mailto:
[hidden email]] On Behalf Of David Knecht charter
Sent: Saturday, April 23, 2011 6:03 PM
To:
[hidden email]
Subject: Re: Live cell imaging with multiphoton confocal
*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy*****
Is there any disadvantage to multiphoton for cultured cells (besides cost and complexity)? Cell viability? Phototoxicity? Dave
On Apr 23, 2011, at 11:21 AM, Kate Luby-Phelps wrote:
> *****
> To join, leave or search the confocal microscopy listserv, go to:
>
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy> *****
>
> Multiphoton has no clear advantage for cells in culture. For cell
> culture samples, we use two photon only to excite DAPI or other UV
> and near-UV fluors since we had to make a choice between the Ti:S and a 405 laser on our scope.
>
> Kate Luby-Phelps
Dr. David Knecht
Department of Molecular and Cell Biology Co-head Flow Cytometry and Confocal Microscopy Facility
U-3125
91 N. Eagleville Rd.
University of Connecticut
Storrs, CT 06269
860-486-2200
860-486-4331 (fax)