Developing photo activated localization microscopy

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4th IEEE International Symposium on Biomedical Imaging: From Nano to Macro. 2007 Apr 15;. doi: 10.1109/isbi.2007.357008

Developing photo activated localization microscopy Hess LabLippincott-Schwartz LabBetzig Lab

Patterson George, Betzig Eric, Lippincott-Schwartz Jennifer, Hess Harald

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Abstract

In conventional biological imaging, diffraction places a limit on the minimal xy distance at which two marked objects can be discerned. Consequently, resolution of target molecules within cells is typically coarser by two orders of magnitude than the molecular scale at which the proteins are spatially distributed. Photoactivated localization microscopy (PALM) optically resolves selected subsets of protect fluorescent probes within cells at mean separations of <25 nanometers. It involves serial photoactivation and subsequent photobleaching of numerous sparse subsets of photoactivated fluorescent protein molecules. Individual molecules are localized at near molecular resolution by determining their centers of fluorescent emission via a statistical fit of their point-spread-function. The position information from all subsets is then assembled into a super-resolution image, in which individual fluorescent molecules are isolated at high molecular densities. In this paper, some of the limitations for PALM imaging under current experimental conditions are discussed.

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Developing photo activated localization microscopy

Abstract

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