SMILE Microscopy : fast and single-plane based super-resolution volume imaging

TL;DR

SMILE microscopy enables rapid, single-plane based super-resolution 3D imaging by encoding molecules across multiple planes, reducing acquisition time and photodamage, and eliminating the need for z-plane scanning.

Contribution

This paper introduces SMILE microscopy, a novel method for simultaneous multi-plane super-resolution imaging that simplifies 3D data acquisition and reduces photobleaching.

Findings

Achieves fast 3D super-resolution imaging without z-plane scanning.

Uses PSF calibration to identify molecules across planes.

Reduces photobleaching and imaging time.

Abstract

Fast 3D super-resolution imaging is essential for decoding rapidly occurring biological processes. Encoding single molecules to their respective planes enable simultaneous multi-plane super-resolution volume imaging. This saves the data-acquisition time and as a consequence reduce radiation-dose that lead to photobleaching and other undesirable photochemical reactions. Detection and subsequent identification of the locus of individual molecule (both on the focal plane and off-focal planes) holds the key. Experimentally, this is achieved by accurate calibration of system PSF size and its natural spread in off-focal planes using sub-diffraction fluorescent beads. Subsequently the identification and sorting of single molecules that belong to different axial planes is carried out (by setting multiple cut-offs to respective PSFs). Simultaneous Multiplane Imaging based Localization Encoded (SMILE) microscopy technique eliminates the need for multiple z-plane scanning and thereby provides a truly simultaneous multiplane super-resolution imaging.