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Near Infrared Diffuse In Vivo Flow Cytometry

View ORCID ProfileJoshua Pace, View ORCID ProfileFernando Ivich, Eric Marple, View ORCID ProfileMark Niedre
doi: https://doi.org/10.1101/2022.05.11.491330
Joshua Pace
aNortheastern University, Department of Bioengineering, Boston, USA
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Fernando Ivich
aNortheastern University, Department of Bioengineering, Boston, USA
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Eric Marple
bEmVision LLC, Loxahatchee, USA
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Mark Niedre
aNortheastern University, Department of Bioengineering, Boston, USA
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  • For correspondence: m.niedre@neu.edu
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Abstract

Significance Diffuse in vivo Flow Cytometry (DiFC) is an emerging technique for enumerating rare fluorescently labeled circulating cells non-invasively 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 In this work, 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 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. This work could pave the way for future use of NIR-DiFC in humans.

Competing Interest Statement

The authors have declared no competing interest.

Footnotes

  • https://github.com/mark-niedre/Design-and-Validation-of-NIR-DiFC/tree/v1.0

  • https://discover.pennsieve.io/datasets/235

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted May 11, 2022.
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Near Infrared Diffuse In Vivo Flow Cytometry
Joshua Pace, Fernando Ivich, Eric Marple, Mark Niedre
bioRxiv 2022.05.11.491330; doi: https://doi.org/10.1101/2022.05.11.491330
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Near Infrared Diffuse In Vivo Flow Cytometry
Joshua Pace, Fernando Ivich, Eric Marple, Mark Niedre
bioRxiv 2022.05.11.491330; doi: https://doi.org/10.1101/2022.05.11.491330

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