Practical Considerations in the Selection and Application of Fluorescent Probes

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Abstract

Due to its sensitivity, multiplexing capacity, and applicability to live specimens, fluorescence is the dominant contrast mechanism used in three-dimensional (3D) biological microscopy. Use of fluorescence detection generally requires specimens to be labeled with extrinsic probes. This is because most biological molecules and structures of interest are not intrinsically fluorescent in spectral ranges that are useful for detection, and even those that are cannot usually be discriminated from each other on the basis of their intrinsic fluorescence.