Robust high-throughput superresolution imaging with speckle illumination

Boston University (Charles River Campus), Boston MA

Investigators

Abstract

ABSTRACT Superresolution (SR) fluorescence microscopy has significantly advanced biological imaging but faces challenges such as the need for specialized molecules, expensive hardware, and long acquisition times. Structured illumination microscopy (SIM) has been successful due to its compatibility with standard fluorescent molecules but requires precise control of fringe patterns. Variations of SIM based on random speckle illumination offer better robustness and easier implementation but suffer from noise and long averaging times. This proposal aims to improve speckle-based SIM by developing new algorithms, engineering speckle illumination, and integrating these into microscopes offering high speed, multicolor and volumetric (multiplane) imaging. Our overall goal is to develop speckle-based SR platforms that provide higher throughput than currently available SR technologies, while being able to image conventional fluorescent molecules with standard labeling protocols. To demonstrate the benefits of speckle-based SR, we will focus on the application of spatial transcriptomics in mouse brain tissue, with the aim of achieving higher resolution and faster overall processing times. Our proposal involves a collaboration between optics and neurobiology labs to establish the technology of speckle-based SR for practical use.