Abstract

The combination of optical sectioning provided by a two-photon temporal focusing with lateral super resolution provided by PALM enables imaging of protein distributions in cells with a lateral localization precision better than 20 nm at multi ple imaging planes over an axial range of 10μm in both mitochondrially labeled fixed cells, and in the membranes of living S2 Drosophila cells. The present super-resolution techniques, such as photoactivated localization microscopy (PALM) or stochastic optical reconstruction microscopy (STORM), do not allow imaging of whole cells and organelles that are typically up to 15 μm deep into the cell, because they are limited to an imaging depth of order to of a fraction of the wavelength. The two photon temporal focusing provides the optical sectioning necessary for a 3D super resolution technique.

© 2010 Optical Society of America