Nature Methods: Light-sheet Fluorescence Microscopy - Method of the Year 2014

Light-sheet Fluorescence Microscopy can Image Living Samples in Three Dimensions with Relatively Low Phototoxicity and at High Speed

January 14, 2015

Just about everyone who has examined fluorescent samples under the microscope is aware of the constant struggle to have enough signal to see the labeled structures while also avoiding fluorophore bleaching. What may be less apparent, at least to those who image bright, robust or fixed samples, is how stressful and potentially toxic to living cells and tissues it is to illuminate them with high-intensity light.

Light-sheet fluorescence microscopy enables relatively gentle imaging of biological samples with high resolution in three dimensions (3D) and over long periods of time. Especially when combined with high-speed cameras, it is fast enough to capture cellular or subcellular dynamics. For its potential for fast, relatively gentle, volumetric imaging of biological samples, we have chosen light-sheet fluorescence microscopy as Method of the Year 2014.

Figure: Comparison of different microscopy illumination modalities (LSFM: lightsheet fluorescence microscopy, WF: widefield microscopy, CF: confocal microscopy). Source: Wikimedia Commons/Jkrieger.

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Method of the Year 2014

Nature Methods 12: 1 (2015); doi: 10.1038/nmeth.3251