Brown, C. M.
Fluorescence microscopy: avoiding the pitfalls.  Journal of Cell Science 120: 1703-1705 (2007).

The advent of fluorescent proteins and the continued development of novel fluorescent probes have put fluorescence microscopy at the center of life science research. Fluorescence microscopes range from relatively straight-forward wide-field microscopes to highly specialised spectral-imaging confocal microscopes. Confocal laser scanning microscopes (LSM) are used to improve fluorescence image quality by eliminating out-of-focus fluorescence and for 3D imaging using software reconstruction. Spinning disk confocal microscopes offer the advantage of video rate (30 frames per second) imaging with charge-coupled device (CCD) cameras. For live cell imaging at these speeds, 3D structures can be imaged on the subsecond time scale with the added benefit of reduced photobleaching/phototoxicity. Programmable array microscopes and line scanning microscopes are available and offer similar advantages to spinning-disk confocals. Finally, multi-photon microscopes use infrared light, which readily penetrates up to 600 μm, allowing deep tissue imaging in living animals.