Fast, multicolour optical sectioning over extended fields of view by combining interferometric SIM with machine learning

Structured illumination can reject out-of-focus signal from a sample, enabling high-speed and high-contrast imaging over large areas with widefield detection optics. Currently, this optical-sectioning technique is limited by image reconstruction artefacts and the need for sequential imaging of multiple colour channels. We combine multicolour interferometric pattern generation with machine-learning processing, permitting high-contrast, real-time reconstruction of image data. The method is insensitive to background noise and unevenly phase-stepped illumination patterns. We validate the method in silico and demonstrate its application on diverse specimens, ranging from fixed and live biological cells to synthetic biosystems, imaging at up to 37 Hz across a 44 x 44 $\mu m^2$ field of view.

An open-source software package for on-the-fly deskewing and live viewing of volumetric lightsheet microscopy data

Oblique plane microscopy, OPM, is a form of lightsheet microscopy that permits volumetric imaging of biological samples at high temporal and spatial resolution. However, the imaging geometry of OPM, and related variants of light sheet microscopy, distorts the coordinate frame of the presented image sections with respect to real space coordinate frame in which the sample is moved to navigate to regions of interest. This makes live viewing and practical operation of such microscopes difficult. We present an open-source software package that utilises GPU acceleration and multiprocessing to transform the display of OPM imaging data in real time to produce live views that mimic that produced by standard widefield microscopes. Image stacks can be acquired, processed and plotted at rates of several Hz, making live operation of OPMs, and similar microscopes, more user friendly and intuitive.