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.
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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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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