Computerized fluorescence microscopic vision in the biomedical sciences

The quantitative information obtainable from light-microscope images has been improved dramatically through the combination of advanced digital image processing technology, novel optical microscope techniques and highly sensitive video cameras. The ability of this instrumentation to intensify weakly fluorescent signals at the level of single living cells, without injuring the cell, has led to major advances in the understanding of the physiology and pathophysiology of living cells. This review discusses the instrumentation required to assemble computerized microscopic vision systems for quantitative fluorescent imaging. In addition, this paper identifies the currently available confocal microscopes for live cell imaging. Four more recently developed types of computerized microscopic vision systems, digitized fluorescence polarization microscopy (DFPM), automated fluorescence image cytometry (AFIC), time-resolved fluorescence lifetime imaging microscopy (TRFLIM), and fluorescence resonance energy transfer microscopy (FRETM) are also discussed. This review paper ends by providing specific examples of biomedical applications such as the role of Ca²⁺ in cell growth, pH alterations during endocytosis, changes in membrane structure that accompany hypoxic injury, and detection of Human Papillomavirus (HPV) infection in clinical cervicovaginal smears which can be undertaken with this type of instrumentation.