Fluorescence microscopy principle

Morgan, C. G., Mitchell, A. C. and Murray, J. G. (1990). Nano-second time-resolved fluorescence microscopy principles and practice. Trans. R. Microsc. Soc. 1, 463-6. 

As shown in Section 11.2.1.1, more details can be obtained by confocal fluorescence microscopy than by conventional fluorescence microscopy. In principle, the extension of conventional FLIM to confocal FLIM using either time- or frequency-domain methods is possible. However, the time-domain method based on singlephoton timing requires expensive lasers with high repetition rates to acquire an image in a reasonable time, because each pixel requires many photon events to generate a decay curve. In contrast, the frequency-domain method using an inexpensive CW laser coupled with an acoustooptic modulator is well suited to confocal FLIM. 

Bacallao, R., Morgane, B., Stelzer, E. H. K., and DeMey, J. (1989) Guiding principles of specimen preservation for confocal fluorescence microscopy, in Handbook of Biological Confocal Microscopy (Pawley, J. B., ed.). Plenum, New York, pp. 197-205. 

Figure 1 Principle of the SNOM/AFM (b) as compared with that of the standard fluorescence microscopy (a).

Bacallao R, Bomsel M, Stelzer EHK, DeMay J (1989) Guiding principles of specimen preservation for confocal fluorescence microscopy. 

Figure 16. Measurement principle of total internal reflection fluorescence microscopy.

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