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Dear List,

I would appreciate if someone would share his/her experience on imaging CFP,
CeFP, ATTO 325 or other similar spectra dyes under TIRF illumination by 430
nm (Rainbow Photonics) or 442 nm (Crystal Laser) laser lines.

We can image CeFP (a very useful peptide by Dave Piston) labeled viruses
largely avoiding photo bleaching in wide-field microscopy (100x NA 1.45,
Orca AG at 2x2 binning, 450 msec exposures, ProlongGold mounting media,
Semrock CFP-YFP Sedat filter set).  

I have observed high photo bleaching rate of YFP labeled viruses (514 nm
Argon line) in TIRFM (1% n-propyl gallate in 1xPBS/5% EthOH). 

Vitaly
NCI-Frederick,
301-846-6575