Fluorescence has long been recognized as a powerful tool for probing biological structure and function. Because probe molecules can be very much more fluorescent than the constituents of most biological specimens, the signal for the exogenous fluorophores can be measured continuously and nondestructively with excellent spatial and temporal resolution in living cells (Waggoner 1 986). The earliest developed and most straightÂ forward uses of fluorescent groups are simply as positional tags or markers. Examples are immunofluorescence labeling (Nairn 1 976), fluorescent analog cytochemistry (Taylor et al 1 986a), vital staining of organelles (Pagano & Sleight 1 985, Wang & Taylor 1 988), assessment of cell morÂ phology or intercellular coupling with microinjected tracers (Stewart 1 98 1 ), measurement of distances between probes by fluorescence energy transfer (Stryer 1978, Uster & Pagano 1 986), and measurement of diffusion coefficients and exchange rates by photobleaching recovery (Elson 1986). The common feature of such applications is that the main role of the fluorescent group is merely to signal its presence and location rather than to sense its environment. The main criteria for such fluorescent tags are simple: wavelengths of excitation and emission, brightness, photostability (Mathies & Stryer 1 986), size
Annual Review of Neuroscience – Annual Reviews
Published: Mar 1, 1989
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