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Jens - in your Traffic study, were the tubulin-fp constructs N terminal or C terminal? Does it make a difference?
Rick
Richard Hallworth, Ph.D., Associate Professor
Director, Nebraska Center for Cell Biology
http://www.biomedsci.creighton.edu/facilities/nccb.html
Department of Biomedical Sciences
Creighton University School of Medicine
2500 California Plaza
Omaha NE 68178
Ph: (402) 280-3057
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-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On Behalf Of Rietdorf, Jens
Sent: Wednesday, January 03, 2007 3:10 AM
To: [log in to unmask]
Subject: Re: live cell
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Dear Mark and others,
(happy new year!)
Be aware, fp-fusions change cytoskeleton dynamics (at least for tubulin). We compared growth and shrinkage rates to other labeling methods.
Colombelli J, Reynaud EG, Rietdorf J, Pepperkok R, Stelzer EH. In vivo selective cytoskeleton dynamics quantification in interphase cells induced by pulsed ultraviolet laser nanosurgery. Traffic. 2005 Dec;6(12):1093-102.
The protein used to label actin (mentioned in other postings) to my knowledge is a construct consisting of lim domains that bind to and thereby indirectly label actin filaments. It would be interesting to see if binding of this construct to the filaments changes actin dynamics.
Fischer, M., Haase, I., Simmeth, E., Gerisch, G. and MuĻ ller-
Taubenberger, A. (2004) A brilliant monomeric red fluorescent
protein to visualize cytoskeleton dynamics in Dictyostelium. FEBS
Lett. 577, 227-232.
To improve visualisation, TIRF is the imaging method of choice for adherent cells. Otherwise use Yokogawa disk.
regards, jens
---
Dr. Jens Rietdorf
head of microscopy unit
Friedrich-Miescher-Institute, wro1066.2.32
Maulbeerstr.66, CH-4058 Basel, Switzerland
phone +41(61)69-75172 mobil +41 798284737
-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On Behalf Of Marc Thibault
Sent: Mittwoch, 20. Dezember 2006 22:23
To: [log in to unmask]
Subject: live cell
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Hello all,
Does anyone one know of commercially available GFP (or other FP's) tagged cells, with a preference for cytoskeletal tagged proteins ?
Thanks
Marc
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