High resolution multiplexed functional imaging in live embryos (Conference Presentation)

Dongli Xu; Weibin Zhou; Leilei Peng

doi:10.1117/12.2252818

19 April 2017 High resolution multiplexed functional imaging in live embryos (Conference Presentation)

Dongli Xu, Weibin Zhou, Leilei Peng

Proceedings Volume 10043, Diagnosis and Treatment of Diseases in the Breast and Reproductive System; 100430M (2017) https://doi.org/10.1117/12.2252818
Event: SPIE BiOS, 2017, San Francisco, California, United States

Abstract

Fourier multiplexed fluorescence lifetime imaging (FmFLIM) scanning laser optical tomography (FmFLIM-SLOT) combines FmFLIM and Scanning laser optical tomography (SLOT) to perform multiplexed 3D FLIM imaging of live embryos. The system had demonstrate multiplexed functional imaging of zebrafish embryos genetically express Foster Resonant Energy Transfer (FRET) sensors. However, previous system has a 20 micron resolution because the focused Gaussian beam diverges quickly from the focused plane, makes it difficult to achieve high resolution imaging over a long projection depth. Here, we present a high-resolution FmFLIM-SLOT system with achromatic Bessel beam, which achieves 3 micron resolution in 3D deep tissue imaging. In Bessel-FmFLIM-SLOT, multiple laser excitation lines are firstly intensity modulated by a Michelson interferometer with a spinning polygon mirror optical delay line, which enables Fourier multiplexed multi-channel lifetime measurements. Then, a spatial light modulator and a prism are used to transform the modulated Gaussian laser beam to an achromatic Bessel beam. The achromatic Bessel beam scans across the whole specimen with equal angular intervals as sample rotated. After tomography reconstruction and the frequency domain lifetime analysis method, both the 3D intensity and lifetime image of multiple excitation-emission can be obtained. Using Bessel-FmFLIM-SLOT system, we performed cellular-resolution FLIM tomography imaging of live zebrafish embryo. Genetically expressed FRET sensors in these embryo will allow non-invasive observation of multiple biochemical processes in vivo.

Conference Presentation

Citation Download Citation

Dongli Xu, Weibin Zhou, and Leilei Peng "High resolution multiplexed functional imaging in live embryos (Conference Presentation)", Proc. SPIE 10043, Diagnosis and Treatment of Diseases in the Breast and Reproductive System, 100430M (19 April 2017); https://doi.org/10.1117/12.2252818

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KEYWORDS

Imaging systems

Multiplexing

Image resolution

3D image processing

Bessel beams

Fluorescence lifetime imaging

Functional imaging

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