Journal
MICROSCOPY AND MICROANALYSIS
Volume 14, Issue 6, Pages 492-506Publisher
CAMBRIDGE UNIV PRESS
DOI: 10.1017/S1431927608080835
Keywords
two-photon excitation; laser scanning microscopy; second harmonic generation; calcium imaging
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Funding
- National Institutes of Health
- American Heart Association
- Showalter Grant from Indiana University
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Two-photon excitation microscopy has become the standard technique for high resolution deep tissue and intravital imaging. It provides intrinsic three-dimensional resolution in combination with increased penetration depth compared to single-photon confocal microscopy. This article will describe the basic physical principles of two-photon excitation and will review its multiple applications to cardiovascular imaging, including second harmonic generation and fluorescence laser scanning microscopy. In particular, the capability and limitations of multiphoton microscopy to assess functional heterogeneity on a cellular scale deep within intact, Langendorff-perfused hearts are demonstrated. It will also discuss the use of two-photon excitation-induced release of caged compounds for the study of intracellular calcium signaling and intercellular dye transfer.
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