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Proceedings Paper

In vivo imaging flow cytometry based on laser scanning two-photon microscopy at kHz cross-sectional frame rate
Author(s): Lingjie Kong; Jianyong Tang; Meng Cui
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Paper Abstract

In vivo flow cytometry has found numerous applications in biology and pharmacology. However, conventional cytometry does not provide the detailed morphological information that is needed to fully determine the phenotype of individual circulating cells. Imaging cytometry, capable of visualizing the morphology and dynamics of the circulating cells at high spatiotemporal resolution, is highly desired. Current wide-field based image cytometers are limited in the imaging depth and provide only two-dimensional resolution. For deep tissue imaging, laser scanning two-photon fluorescence microscopy (TPM) is widely adopted. However, for applications in flow cytometry, the axial scanning speed of current TPMs is inadequate to provide high-speed cross-sectional imaging of vasculature. We have integrated an optical phase-locked ultrasound lens into a standard TPM and achieved microsecond-scale axial scanning. With a galvo scanner for transverse scanning, we achieved kHz cross-sectional frame rate. Here we report its applications for in vivo deformability cytometry and in vivo imaging flow cytometry, and demonstrate the capability of imaging dynamical morphologies of flowing cells, distinguishing cells and cellular clusters, and simultaneously quantifying different cell populations based on their fluorescent labels.

Paper Details

Date Published: 14 March 2016
PDF: 6 pages
Proc. SPIE 9712, Multiphoton Microscopy in the Biomedical Sciences XVI, 971203 (14 March 2016); doi: 10.1117/12.2211418
Show Author Affiliations
Lingjie Kong, Purdue Univ. (United States)
Jianyong Tang, National Institutes of Health (United States)
Meng Cui, Purdue Univ. (United States)

Published in SPIE Proceedings Vol. 9712:
Multiphoton Microscopy in the Biomedical Sciences XVI
Ammasi Periasamy; Peter T. C. So; Karsten König, Editor(s)

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