Mr. Jongha Lee Profile

Jongha Lee

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Proceedings Article | 12 March 2024 Paper

Thermal investigation for determining safe laser power limits in two-photon line excitation array detection microscopy

Aditya Roy, Biswajit Mishra, Daniel Wu, Jongha Lee, Adela Ben-Yakar

Proceedings Volume 12847, 128470M (2024) https://doi.org/10.1117/12.3002191

KEYWORDS: Simulations, Brain tissue, Tissues, Laser safety, Monte Carlo methods, Numerical simulations, Detector arrays, Two photon imaging, Sampling rates, Natural surfaces

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With the aim of advancing modern neuroscience and sampling neurons at up to 100 kHz frame rates, our group is developing a novel Two-photon Line Excitation Array Detection (2p-LEAD) imaging modality. Performing high resolution two-photon imaging at such high sampling rates necessitates the deposition of a large number of photons within the focal volume, which in turn warrants high laser powers. Consequently, the risk of heating and thermal damage limits the imaging speed and depth. In contrast to point-scan two-photon imaging, where safe average laser power values of 200 mW with conventional objective cooling have been established, there are no thermal characterization studies in the case of line-scan imaging modalities that could enable us in determining maximum laser powers to prevent tissue heating damage. We recently demonstrated through numerical investigations that enhanced cooling strategies of imparting laminar flow to the objective immersion water layer while implementing laser duty cycles could potentially increase safe power levels up to 600 mW of average surface power in the case of point scanning. A clear understanding of the effects of laser dosimetry on optical parameters of line-scan systems is essential to determine safe power values that would prevent thermal damage. In this work, we perform 3D MC-FDM numerical simulations at 1035 nm wavelength with a novel beam focusing framework over a parameter space spanning average powers and imaging depth to predict optothermal interactions. With experimental validation studies on tissue phantoms, our work would establish a much-needed power threshold in two-photon line scanning, which is an emerging modality of choice for high-speed volumetric imaging systems.

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