Two-photon flow cytometer with laser scanning Bessel beams

Yongdong Wang; Yu Ding; Supriyo Ray; Aurelio Paez; Chuan Xiao; Chunqiang Li

doi:10.1117/12.2208549

21 March 2016 Two-photon flow cytometer with laser scanning Bessel beams

Yongdong Wang, Yu Ding, Supriyo Ray, Aurelio Paez, Chuan Xiao, Chunqiang Li

Proceedings Volume 9709, Biophotonics and Immune Responses XI; 97090F (2016) https://doi.org/10.1117/12.2208549
Event: SPIE BiOS, 2016, San Francisco, California, United States

Abstract

Flow cytometry is an important technique in biomedical discovery for cell counting, cell sorting and biomarker detection. In vivo flow cytometers, based on one-photon or two-photon excited fluorescence, have been developed for more than a decade. One drawback of laser beam scanning two-photon flow cytometer is that the two-photon excitation volume is fairly small due to the short Rayleigh range of a focused Gaussian beam. Hence, the sampling volume is much smaller than one-photon flow cytometry, which makes it challenging to count or detect rare circulating cells in vivo. Bessel beams have narrow intensity profiles with an effective spot size (FWHM) as small as several wavelengths, making them comparable to Gaussian beams. More significantly, the theoretical depth of field (propagation distance without diffraction) can be infinite, making it an ideal solution as a light source for scanning beam flow cytometry. The trade-off of using Bessel beams rather than a Gaussian beam is the fact that Bessel beams have small concentric side rings that contribute to background noise. Two-photon excitation can reduce this noise, as the excitation efficiency is proportional to intensity squared. Therefore, we developed a two-photon flow cytometer using scanned Bessel beams to form a light sheet that intersects the micro fluidic channel.

Conference Presentation

Citation Download Citation

Yongdong Wang, Yu Ding, Supriyo Ray, Aurelio Paez, Chuan Xiao, and Chunqiang Li "Two-photon flow cytometer with laser scanning Bessel beams", Proc. SPIE 9709, Biophotonics and Immune Responses XI, 97090F (21 March 2016); https://doi.org/10.1117/12.2208549

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