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Determining the Depth Limit of Bioluminescent Sources in Scattering Media

View ORCID ProfileAnkit Raghuram, Fan Ye, Jesse K. Adams, Nathan Shaner, Jacob T. Robinson, Ashok Veeraraghavan
doi: https://doi.org/10.1101/2020.04.21.044982
Ankit Raghuram
1Department of Electrical and Computer Engineering, Rice University, Houston, TX 77005, USA
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  • For correspondence: ar89@rice.edu
Fan Ye
1Department of Electrical and Computer Engineering, Rice University, Houston, TX 77005, USA
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Jesse K. Adams
1Department of Electrical and Computer Engineering, Rice University, Houston, TX 77005, USA
2Applied Physics Program, Rice University, Houston, TX 77005, USA
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Nathan Shaner
3Department of Neurosciences, University of California San Diego, La Jolla, CA 92093, USA
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Jacob T. Robinson
1Department of Electrical and Computer Engineering, Rice University, Houston, TX 77005, USA
2Applied Physics Program, Rice University, Houston, TX 77005, USA
4Department of Neuroscience, Baylor College of Medicine, Houston, TX 77030, USA
5Department of Bioengineering, Rice University, Houston, TX 77005, USA
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Ashok Veeraraghavan
1Department of Electrical and Computer Engineering, Rice University, Houston, TX 77005, USA
2Applied Physics Program, Rice University, Houston, TX 77005, USA
6Department of Computer Science, Rice University, Houston TX 77005, USA
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Abstract

Bioluminescence has several potential advantages compared to fluorescence microscopy for in vivo biological imaging. Because bioluminescence does not require excitation light, imaging can be performed for extended periods of time without phototoxicity or photobleaching, and optical systems can be smaller, simpler, and lighter. Eliminating the need for excitation light may also affect how deeply one can image in scattering biological tissue, but the imaging depth limits for bioluminescence have yet to be reported. Here, we perform a theoretical study of the depth limits of bioluminescence microscopy and find that cellular resolution imaging should be possible at a depth of 5-10 mean free paths (MFPs). This limit is deeper than the depth limit for confocal microscopy and slightly lower than the imaging limit expected for two-photon microscopy under similar conditions. We also validate our predictions experimentally using tissue phantoms. Overall we show that with advancements in the brightness of bioluminescent indicators, it should be possible to achieve deep, long-term imaging in biological tissue with cellular resolution.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
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Posted April 23, 2020.
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Determining the Depth Limit of Bioluminescent Sources in Scattering Media
Ankit Raghuram, Fan Ye, Jesse K. Adams, Nathan Shaner, Jacob T. Robinson, Ashok Veeraraghavan
bioRxiv 2020.04.21.044982; doi: https://doi.org/10.1101/2020.04.21.044982
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Determining the Depth Limit of Bioluminescent Sources in Scattering Media
Ankit Raghuram, Fan Ye, Jesse K. Adams, Nathan Shaner, Jacob T. Robinson, Ashok Veeraraghavan
bioRxiv 2020.04.21.044982; doi: https://doi.org/10.1101/2020.04.21.044982

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