fastSIM: a practical implementation of fast structured illumination microscopy

HW Lu-Walther, M Kielhorn, R Förster… - … and Applications in …, 2015 - iopscience.iop.org
HW Lu-Walther, M Kielhorn, R Förster, A Jost, K Wicker, R Heintzmann
Methods and Applications in Fluorescence, 2015iopscience.iop.org
A significant improvement in acquisition speed of structured illumination microscopy (SIM)
opens a new field of applications to this already well-established super-resolution method
towards 3D scanning real-time imaging of living cells. We demonstrate a method of
increased acquisition speed on a two-beam SIM fluorescence microscope with a lateral
resolution of~ 100 nm at a maximum raw data acquisition rate of 162 frames per second
(fps) with a region of interest of 16.5× 16.5 µm 2, free of mechanically moving components …
Abstract
A significant improvement in acquisition speed of structured illumination microscopy (SIM) opens a new field of applications to this already well-established super-resolution method towards 3D scanning real-time imaging of living cells. We demonstrate a method of increased acquisition speed on a two-beam SIM fluorescence microscope with a lateral resolution of~ 100 nm at a maximum raw data acquisition rate of 162 frames per second (fps) with a region of interest of 16.5× 16.5 µm 2, free of mechanically moving components. We use a programmable spatial light modulator (ferroelectric LCOS) which promises precise and rapid control of the excitation pattern in the sample plane. A passive Fourier filter and a segmented azimuthally patterned polarizer are used to perform structured illumination with maximum contrast. Furthermore, the free running mode in a modern sCMOS camera helps to achieve faster data acquisition.
iopscience.iop.org
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