Aberration free refocusing for high numerical aperture microscopy - art. no. 686110

In this paper, we present a novel technique that permits sectioning microscopes to refocus and acquire images from a large range of specimen depths without introducing movements near the specimen. In contrast to other such remote focusing methods, this technique avoids systematic aberrations that degrade image quality when imaging planes away from the optimal focal plane. Furthermore, the specific geometry that is employed in this work enables refocusing to be carried out at high speed and hence permits, for the first time, a number of dynamical biological processes to be observed. Although this technique can be applied to any optical imaging system, it is particularly suited to the case of high numerical aperture microscope systems. To demonstrate this we present results from two prototype systems built in our laboratory based on a slit scanning confocal fluorescence microscope and a two photon fluorescence microscope.