Multiphoton microscopy: in depth, in vivo sub cellular resolution imaging

Girkin, J.M. (2006) Multiphoton microscopy: in depth, in vivo sub cellular resolution imaging. Proceedings of SPIE: The International Society for Optical Engineering, 6163. 0T1 - 0T10. (http://www.10.1117/12.697070)

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Since the advent of the optical microscope around 400 years ago there has been an increasing desire by life science researchers to image ever more deeply into samples with high resolution. More recently this desire has lead to the requirement to image three dimensional, living samples with sub-cellular resolution. The application of practical confocal microscopes partially solved this challenge but it was not until the development of multiphoton imaging methods 15 years ago that true in vivo, imaging with high resolution could take place at depth within samples. This paper reviews the basic principles behind multiphoton microscopy and the advances that have been made in the last five years with regard to real time, in depth, imaging. Consideration is given to the best design of multiphoton instruments along with recent research that has been undertaken in the use of active optical elements to enhance in vivo imaging. Some guidelines for the correct selection of the microscopy method for a range of life science challenges are also presented.