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Single Molecule Light Field Microscopy

Sims, R. R.; Abdul Rehman, S.; Lenz, M. O.; Clark, A.; Sanders, E. W.; Ponjavic, A.; Muresan, L.; Lee, S. F.; O'Holleran, K.

2020-05-22 biophysics
10.1101/2020.05.20.104802 bioRxiv
Show abstract

We introduce single molecule light field microscopy (SMLFM), a novel 3D single molecule localization technique that is capable of up to 20 nm isotropic precision across a 6 m depth of field. SMLFM can be readily implemented by installing a refractive microlens array into the conjugate back focal plane of any widefield single molecule localization system. We demonstrate that 3D localization can be performed by post-processing 2D localization data generated by common, widely-used, algorithms. In this work we benchmark the performance of SMLFM and finally showcase its capabilities by imaging fluorescently labeled membranes of fixed eukaryotic cells below the diffraction limit.

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