Fast, 3D imaging via confocal line scanning of a Bessel beam using a single galvo mirror

Pengfei Zhang; Peter M. Goodwin; James H. Werner

doi:10.1117/12.2036848

4 March 2014 Fast, 3D imaging via confocal line scanning of a Bessel beam using a single galvo mirror

Pengfei Zhang, Peter M. Goodwin, James H. Werner

Proceedings Volume 8947, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XII; 89471K (2014) https://doi.org/10.1117/12.2036848
Event: SPIE BiOS, 2014, San Francisco, California, United States

Abstract

We developed a light-sheet illumination microscope that can perform fast 3D imaging of transparent biological samples. The light-sheet is created by raster scanning of a Bessel Beam with one galvo-mirror. Fluorescence excited from the thin layer of the sample is de-scanned by the same galvo-mirror, and then spatially filtered by a slit so that out-of-focus fluorescence generated by the side lobes of the Bessel beam is rejected. The spatially filtered fluorescence is returned by a series of optics and is re-scanned by the same galvo-mirror across the chip of a camera such that the fluorescence image is constructed in real-time. Compared to two-photon Bessel beam excitation or other confocal line scanning approaches, our method is of lower cost, simpler, and doesn’t require calibration and synchronization of multiple galvo mirrors. We demonstrated the capability of fast 3D imaging and background rejection capabilities of this microscope with fluorescent beads embedded in PDMS.

Citation Download Citation

Pengfei Zhang, Peter M. Goodwin, and James H. Werner "Fast, 3D imaging via confocal line scanning of a Bessel beam using a single galvo mirror", Proc. SPIE 8947, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XII, 89471K (4 March 2014); https://doi.org/10.1117/12.2036848

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