Ratiometric widefield imaging with spectrally balanced detection

Shimon Yudovich, Lior Shani, Asaf Grupi, Omri Bar-Elli, Dan Steinitz, Dan Oron, Shimon Weiss

Research output: Contribution to journalArticlepeer-review


Ratiometric imaging is an invaluable tool for quantitative microscopy, allowing for robust detection of FRET, anisotropy, and spectral shifts of nano-scale optical probes in response to local physical and chemical variations such as local pH, ion composition, and electric potential. In this paper, we propose and demonstrate a scheme for widefield ratiometric imaging that allows for continuous tuning of the cutoff wavelength between its two spectral channels. This scheme is based on angle-tuning the image splitting dichroic beamsplitter, similar to previous works on tunable interference filters. This configuration allows for ratiometric imaging of spectrally heterogeneous samples, which require spectral tunability of the detection path in order to achieve good spectrally balanced ratiometric detection.

Original languageEnglish
Pages (from-to)5385-5394
Number of pages10
JournalBiomedical Optics Express
Issue number10
Publication statusPublished - Oct 1 2019

ASJC Scopus subject areas

  • Biotechnology
  • Atomic and Molecular Physics, and Optics

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