Hello Group,
I have acquired a series of time lapse data sets and am beginning some
simple analysis. The biology I am looking at is accumulation of an
EGFP-tagged protein at the site of UV-laser induced DNA damage. The images
were acquired using an EM-CCD and neutral density filter attenuated
widefield illumination coming from an EXFO 100W Hg lamp. The EGFP signal
accumulates in a well defined spot and I am able analyze the rate of
accumulation and fold-increase of florescence intensity at several
same-sized ROI's using Image J plugins. Baseline fluorescence intensity
subtraction is simple enough. I have used a non-lased cell in the same field
of view to monitor EGFP photo-bleaching, calculated the slope for this, but
am stuck on how I might use this information to develop a correction factor.
I would appreciate any thoughts and help in this regard.
Thanks to all.
Farid

-- 
Farid Jalali MSc
Program Leader- Cellular Imaging Core
Applied Molecular Oncology and Radiation Medicine Program
Princess Margaret Hospital (University  Health Network)
Toronto Medical Discovery Tower
Toronto, Canada
416-581-7754 STTARR at TMDT
416-581-7791 STTARR Microscopy Suite