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bioRxiv · 10.64898/2026.03.02.709062

Beyond the skin barrier: optical clearing enables non-invasive cortex-wide optical coherence angiography in mice in-vivo

Seong, D.·Yun, S.·Han, S.·Biswas, S.·Kim, B.·Remlova, E.·Razansky, D.·Kim, J.·Ou, Z.·Jeon, M.
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Abstract

Non-invasive, high-resolution visualization of mouse brain vasculature remains challenging due to significant light scattering and absorption by mammalian tissues, hence many optical imaging protocols require scalp and/or skull excision. Here we present a fully reversible tartrazine-based optical clearing strategy that enables cortex-wide optical coherence tomography angiography (OCTA) through intact scalp and skull. We characterized tartrazine properties in the near infrared (NIR)-II band of the 1.3 {micro}m swept-source OCTA system, confirming minimal absorption across 1.25-1.35 {micro}m wavelength range and an effectively constant refractive index, suggesting negligible OCTA distortions. Spatially selective agent application showed that intracranial vessels emerge selectively within the treated region of interest (ROI), whereas untreated regions retain strong interference by the scalp vascular features. Depth-encoded projections and cross-sectional OCTA demonstrated an increased signal recovery at depth and an extended vessel-detection range after clearing. Vessel-map changes were quantified using intersection-over-union and Dice coefficients, yielding high similarity outside the ROI and reduced similarity within the ROI, consistent with a transition from scalp to brain vasculature. Reproducibility was confirmed in three independent 11-week-old mice and validated against scalp-removed reference OCTA. Screening tartrazine in the 0.3-0.8 Molar concentration range (7-min application) identified 0.6 M as optimal for whole-cortex scanning, balancing clearing efficacy and solution handling. Finally, the protocol generalized across mice aged 5-18 weeks. This approach provides a practical route to non-invasive structural cerebrovascular mapping with OCTA.

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Seong, D., Yun, S., Han, S., Biswas, S., Kim, B., Remlova, E., Razansky, D., Kim, J., Ou, Z., Jeon, M.. 2026-03-05. Beyond the skin barrier: optical clearing enables non-invasive cortex-wide optical coherence angiography in mice in-vivo. https://doi.org/10.64898/2026.03.02.709062

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