Abstract
Various techniques have been developed to measure the 2D and 3D
positions and 2D and 3D orientations of fluorescent molecules with
improved precision over standard epifluorescence microscopes. Due to
the challenging signal-to-background ratio in typical single-molecule
experiments, it is essential to choose an imaging system optimized for
the specific target sample. In this work, we compare the performance
of multiple state-of-the-art and commonly used methods for orientation
localization microscopy against the fundamental limits of measurement
precision. Our analysis reveals optimal imaging methods for various
experiment conditions and sample geometries. Interestingly, simple
modifications to the standard fluorescence microscope exhibit superior
performance in many imaging scenarios.
© 2021 Optical Society of
America
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