UNIVERSITY OF MASSACHUSETTS
We will conduct two studies that will serve to improve and enable the accomplishment of the specific aims of 5R01 GM030758: 28-29. The first addresses time and data loss due to human cells migrating off the fields of view during the time lapse observations used for all specific aims. We will make and post on the web a series of 20-63X phase contrast and differential interference contrast time lapse movies of normal human cells during interphase and mitosis. These movies will be used to adapt an existing fluorescent cell tracking algorithm for transmitted light applications. This will allow a motorized microscope stage to keep individual human cells centered in the field of view as they migrate. The movies will also be available to computer scientists to be used in developing algorithms for pattern recognition and analysis. The second project addresses the phototoxicity of the 488nm light used to excite the GFP in many applications with live cells. We will test reports that short (a few milliseconds) but intense bursts of blue light are less damaging to the cells than longer but less intense exposures.