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Multi-photon excitation imaging of dynamic processes in living cells and tissues

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Reviews of Physiology Biochemistry and Pharmacology

Part of the book series: Reviews of Physiology, Biochemistry and Pharmacology ((REVIEWS,volume 160))

Abstract

Over the past decade, two-photon microscopy has successfully made the transition from the laser laboratory into a true biological research setting. This has been due in part to the recent development of turnkey ultrafast laser systems required for two-photon microscopy, allowing ease of use in nonspecialist laboratories. The advantages of two-photon microscopy over conventional optical sectioning techniques are for greater imaging depths and reduced overall phototoxicity, as such enabling noninvasive intra-vital imaging of cellular and subcellular processes. Greater understanding of these advantages has allowed this technique to be more effectively utilized in a biological research setting. This review will cover the recent widespread uses of two-photon microscopy and highlight the wide range of physiological studies enabled in fields such as neurosciences, developmental biology, immunology, cancer biology, and endocrinology.

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Notes

  1. 1.

    Technology such as that used on the LSM5Live line scanning microscope: www.zeiss.com/micro.

Abbreviations

2PM:

Two-photon microscopy

fl:

Femtoliter

fs:

Femtosecond

NA:

Numerical aperture

NDD:

Non-descanned detection

GFP:

Green fluorescent protein

[Ca2+]i :

Intra-cellular calcium concentration

DCs:

Dendritic cells

QD:

Quantum dot

NAD(P)H:

β-Nicotinamide adenine dinucleotide (phosphate)

SHG:

Second harmonic generation

THG:

Third harmonic generation

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Acknowledgements

We are grateful to Gert-Jan Kremers for helpful comments on the manuscript. The authors acknowledge financial support from National Institute of Health grants R01-DK53434, P20-GM72048 and the Medical Free-Electron Laser Program of the United States Department of Defense.

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Authors and Affiliations

  1. Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN, USA

    R. K. P. Benninger, M. Hao & D. W. Piston

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  1. R. K. P. Benninger
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Correspondence to R. K. P. Benninger .

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Benninger, R.K.P., Hao, M., Piston, D.W. (2008). Multi-photon excitation imaging of dynamic processes in living cells and tissues. In: Reviews of Physiology Biochemistry and Pharmacology. Reviews of Physiology, Biochemistry and Pharmacology, vol 160. Springer, Berlin, Heidelberg. https://doi.org/10.1007/112_2008_801

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