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16 September 2022 Near-infrared diffuse in vivo flow cytometry
Joshua Pace, Fernando Ivich, Eric T. Marple, Mark Niedre
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Journal of Biomedical Optics, Vol. 27, Issue 9, 097002 (September 2022). https://doi.org/10.1117/1.JBO.27.9.097002
Abstract

Significance

Diffuse in vivo flow cytometry (DiFC) is an emerging technique for enumerating rare fluorescently labeled circulating cells noninvasively in the bloodstream. Thus far, we have reported red and blue-green versions of DiFC. Use of near-infrared (NIR) fluorescent light would in principle allow use of DiFC in deeper tissues and would be compatible with emerging NIR fluorescence molecular contrast agents.

Aim

We describe the design of a NIR-DiFC instrument and demonstrate its use in optical flow phantoms in vitro and in mice in vivo.

Approach

We developed an improved optical fiber probe design for efficient collection of fluorescence from individual circulating cells and efficient rejection of instrument autofluorescence. We built a NIR-DiFC instrument. We tested this with NIR fluorescent microspheres and cell lines labeled with OTL38 fluorescence contrast agent in a flow phantom model. We also tested NIR-DiFC in nude mice injected intravenously with OTL38-labeled L1210A cells.

Results

NIR-DiFC allowed detection of circulating tumor cells (CTCs) in flow phantoms with mean signal-to-noise ratios (SNRs) of 19 to 32 dB. In mice, fluorescently labeled CTCs were detectable with mean SNR of 26 dB. NIR-DiFC also exhibited orders significantly lower autofluorescence and false-alarm rates than blue-green DiFC.

Conclusions

NIR-DiFC allows use of emerging NIR contrast agents. Our work could pave the way for future use of NIR-DiFC in humans.
CC BY: © The Authors. Published by SPIE under a Creative Commons Attribution 4.0 International License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.
Citation Download Citation
Joshua Pace, Fernando Ivich, Eric T. Marple, and Mark Niedre "Near-infrared diffuse in vivo flow cytometry," Journal of Biomedical Optics 27(9), 097002 (16 September 2022). https://doi.org/10.1117/1.JBO.27.9.097002
Received: 11 May 2022; Accepted: 7 September 2022; Published: 16 September 2022
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Cited by 7 scholarly publications.
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KEYWORDS
In vivo imaging
Near infrared
Signal to noise ratio
Flow cytometry
Luminescence
Blood
Tissues
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