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Revealing Nanoscale Membrane Curvature with Polarized Localization Microscopy

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Abstract

Many biological processes depend on nanoscale membrane curvature at length scales shorter than then the diffraction limit (~200 nm). The orientation of biological membranes has been detected with sub-diffraction-limited resolution via the combination of polarized total internal reflection fluorescence microscopy and fluorescence localization microscopy. Indocarbocyanine dyes, such as DiI, maintain their fluorescence dipole moment in the plane of the membrane and can blink for single fluorophore lateral localization with <20 nm uncertainty and membrane-orientation dependent excitation. The sequential images of individual blinking fluorophores and computational determination of the center of each point spread function enables super-resolution image reconstruction with either p- or s-polarized excitation for resolving nanoscale membrane curvature at unprecedented curvatures.

© 2015 Optical Society of America

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