Modern advances in microscopy: from animal to atom.
The ultimate goal of the modern physiologist is to understand and integrate the biochemistry, biophysics and physiology of undisturbed living cells. To accomplish this goal requires the development and application of approaches that can non-invasively probe a variety of dynamic cellular processes. Although no technique is truly non-invasive, recently developed methods are approaching this objective. Fluorescence has become the most popular optical tool for measuring the ion properties inside living cells, due to sensitivity, specificity, and spatial and temporal resolution. All of these systems use the same basic principle, i.e., the cell or tissue is exposed to a concentrated specific beam of excitation light and the resultant fluorescent signal is detected on a collection device. Photomultiplier tube systems will give maximal temporal resolution. Intensified video camera systems allow for maximal spatial resolution, and simultaneous measurement of fields of cells. Improved spatial resolution can be obtained using confocal microscopes providing both 2D and 3D molecular distribution and activity maps from within cells. In addition to obtaining spatial information from within cells, it is now possible to measure dynamic surface morphometric changes with Angstrom level resolution using the technique of Atomic Force Microscopy (AFM). This review will summarize the different uses of both conventional and nonconventional microscopy to examine dynamic changes in living cells.
Geibel
J
J
Department of Surgery and Cellular and Molecular, Yale University, New Haven, CT 06510-8026.
D002477:Cells; D007202:Indicators and Reagents; D008856:Microscopy, Fluorescence / Q000379:methods; D055095:Optics and Photonics
D007202:Indicators and Reagents