Integrating fluorescence computed tomography with optical sheet illumination for imaging of live single cells

Bin Cao, Rishabh Shetty, Dean Smith, Laimonas Kelbauskas, Deirdre Meldrum

Research output: Contribution to journalArticle

Abstract

We present a new approach for three-dimensional (3D) live single-cell imaging with isotropic sub-micron spatial resolution using fluorescence computed tomography (fCT). A thin, highly inclined and laminated optical (HILO) sheet of light is used for fluorescence excitation in live single cells that are rotated around an axis perpendicular to the optical axis. During a full rotation, 400-500 two-dimensional (2D) projection images of the cell are acquired from multiple viewing perspectives by rapidly scanning the HILO light sheet along the optical axis. We report technical characteristics of the HILO approach and the results of a quantitative comparison with conventional epi fCT, demonstrating that HILO fCT offers significantly (about 17 times) reduced photobleaching and a two-fold improvement in 3D imaging contrast. We discuss potential application areas of the method for cell structure studies in live single cells with isotropic 3D spatial resolution.

Original languageEnglish (US)
Pages (from-to)24020-24030
Number of pages11
JournalOptics Express
Volume26
Issue number18
DOIs
StatePublished - Sep 3 2018

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tomography
illumination
fluorescence
cells
spatial resolution
projection
scanning
excitation

ASJC Scopus subject areas

  • Atomic and Molecular Physics, and Optics

Cite this

Integrating fluorescence computed tomography with optical sheet illumination for imaging of live single cells. / Cao, Bin; Shetty, Rishabh; Smith, Dean; Kelbauskas, Laimonas; Meldrum, Deirdre.

In: Optics Express, Vol. 26, No. 18, 03.09.2018, p. 24020-24030.

Research output: Contribution to journalArticle

Cao, Bin ; Shetty, Rishabh ; Smith, Dean ; Kelbauskas, Laimonas ; Meldrum, Deirdre. / Integrating fluorescence computed tomography with optical sheet illumination for imaging of live single cells. In: Optics Express. 2018 ; Vol. 26, No. 18. pp. 24020-24030.
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