Instrumentation Design for Study of Metabolic Control in Living Cells

Abstract

For the study of living cells in culture using the fluorescence microscope it is important to make provision for the widest possible instrumental armamentarium, including real-time parameter interactive experimental control (RIPEC). In an ideal apparatus (Kohen et al 1991) one might include, for example, (Figure 1) provision for excitation and emission fluorescence spectra, video-optical phase visualization with a long working-distance phase condenser (Hirschberg 1990), scanning near-field microscopy (SNOM), total internal reflection microscopy (TIRM), confocal fluorescence optics, and modulated laser excitation for fluorescent lifetime measurements. The living cells must be contained in a special chamber with regulated temperature and humidity and with sufficient working distance to facilitate micromanipulation and micro-injection. The resulting data are to be displayed in real time, used for computer controlled interactive experimentation, and stored, all by a dedicated computer