Simultaneous multicolor fluorescence imaging using PSF splitting

Nat Methods. 2024 Oct;21(10):1801-1805. doi: 10.1038/s41592-024-02383-7. Epub 2024 Sep 6.

Abstract

We present a way to encode more information in fluorescence imaging by splitting the original point spread function (PSF), which offers broadband operation and compatibility with other PSF engineering modalities and existing analysis tools. We demonstrate the approach using the 'Circulator', an add-on that encodes the fluorophore emission band into the PSF, enabling simultaneous multicolor super-resolution and single-molecule microscopy using essentially the full field of view.

MeSH terms

  • Algorithms
  • Fluorescent Dyes / chemistry
  • Humans
  • Image Processing, Computer-Assisted / methods
  • Microscopy, Fluorescence* / methods
  • Optical Imaging / methods
  • Single Molecule Imaging / methods

Substances

  • Fluorescent Dyes