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Nonlinear magic: multiphoton microscopy in the biosciences

Nature Biotechnology volume 21, pages 1369–1377 (2003)Cite this article

Abstract

Multiphoton microscopy (MPM) has found a niche in the world of biological imaging as the best noninvasive means of fluorescence microscopy in tissue explants and living animals. Coupled with transgenic mouse models of disease and 'smart' genetically encoded fluorescent indicators, its use is now increasing exponentially. Properly applied, it is capable of measuring calcium transients 500 μm deep in a mouse brain, or quantifying blood flow by imaging shadows of blood cells as they race through capillaries. With the multitude of possibilities afforded by variations of nonlinear optics and localized photochemistry, it is possible to image collagen fibrils directly within tissue through nonlinear scattering, or release caged compounds in sub-femtoliter volumes.

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Figure 1: Publications employing, developing or reviewing MPM (from PubMed and ISI).
Figure 2: Localization of excitation by two-photon excitation.
Figure 3: Two-photon action cross-sections.
Figure 4: The two-photon excitation volume.
Figure 5: Components of a multiphoton microscope.
Figure 6: Applications showing various capabilities of MPM.
Figure 7: Two-photon fluorescence and SHG.

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  1. School of Applied and Engineering Physics, 212 Clark Hall, Cornell University, Ithaca, 14853, New York, USA

    Warren R Zipfel, Rebecca M Williams & Watt W Webb

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Zipfel, W., Williams, R. & Webb, W. Nonlinear magic: multiphoton microscopy in the biosciences. Nat Biotechnol 21, 1369–1377 (2003). https://doi.org/10.1038/nbt899

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