Staining and Computerized Viedo Microscopy for Apoptosis

Normal cell death regulates tumor and normal tissue kinetics, and its suppression may beassociated with prognosis and diagnosis of cancer. Apoptosis is the normal physiological process ofselective cell elimination entailing preprogrammed responses to physiological inducers and to a widespectrum of chemotherapeutic drugs. A molecular hallmark of apoptosis is that genomic DNA isfragmented and remains inside the nucleus. For staining 3'-OH ends of the DNA strand breaks in situ,terminal deoxynucleotidyl transferase (TdT) and an affinity-tagged nucleotide are applied directly to amicroscope slide. The chain is extended, and then a detectable antibody conjugate is bound. The PhaseI aims to significantly advance the technology for in situ measurement of apoptosis by (1) developingfaster and more sensitive methods of conferring color on DNA strand breaks in situ, and (2) usingcomputerized data acquisition, digital confocal microscopy, and color image analysis to very rapidlytranslate staining into a quantitative apoptotic labeling index that is directly proportional to tumor cellkinetic rate. In Phase II research, the potential clinical utility of applying this system to answer medicalquestions about cancer treatment, diagnosis and prognosis will be assessed.