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Optical sectioning microscopy with planar or structured illumination

Abstract

A key requirement for performing three-dimensional (3D) imaging using optical microscopes is that they be capable of optical sectioning by distinguishing in-focus signal from out-of-focus background. Common techniques for fluorescence optical sectioning are confocal laser scanning microscopy and two-photon microscopy. But there is increasing interest in alternative optical sectioning techniques, particularly for applications involving high speeds, large fields of view or long-term imaging. In this Review, I examine two such techniques, based on planar illumination or structured illumination. The goal is to describe the advantages and disadvantages of these techniques.

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Figure 1: PIM configurations.
Figure 2: Demonstration of high-speed volumetric imaging with 2P-PIM.
Figure 3: Demonstrations of long-term developmental imaging with PIM.
Figure 4: Demonstration of anatomical imaging with 1P-PIM operated in bidirectional widefield illumination mode.
Figure 5: Schematics of different optical sectioning configurations with structured illumination.
Figure 6: Molecular Probes fluorescently labeled mouse intestine slide imaged with grid-illumination DSD microscopy.
Figure 7: Confocal versus HiLo microscopy.

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Acknowledgements

I acknowledge the help of W. Supatto, K. Chu, D. Lim and T. Ford for critically reading this manuscript.

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Correspondence to Jerome Mertz.

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Mertz, J. Optical sectioning microscopy with planar or structured illumination. Nat Methods 8, 811–819 (2011). https://doi.org/10.1038/nmeth.1709

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