Photoswitching-Enabled Contrast Enhancement in Light Sheet Fluorescence Microscopy

Tom Vettenburg, Angélica Corral, Alberto Rodríguez-Pulido, Cristina Flors, Jorge Ripoll

Research output: Contribution to journalArticle

3 Citations (Scopus)
136 Downloads (Pure)

Abstract

Light sheet fluorescence microscopy enables high-resolution imaging of thick biological samples. By restricting the fluorescence excitation to a single plane, rapid wide-field image acquisition is possible with minimal sample exposure. Although light sheet microscopy is able to resolve subcellular features at depth in model organisms, elevated levels of endogenous autofluorescence often preclude acceptable contrast and may obscure features of interest in general samples. Here we demonstrate how photoswitchable fluorophores can be exploited to boost contrast in light sheet microscopy. The novel detection method enables high specificity while maintaining the optical sectioning capability of the light sheet microscope. Our experiments reveal structures hidden well below the ambient fluorescent background level by enhancing the contrast by 2 orders of magnitude.

Original languageEnglish
Pages (from-to)424-428
Number of pages5
JournalACS Photonics
Volume4
Issue number3
DOIs
Publication statusPublished - 15 Mar 2017

Keywords

  • autofluorescence
  • intensity evolution
  • optical lock-in detection
  • photoswitching
  • selective plane illumination microscopy

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