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June 28th, 2025
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The University of Hong Kong
bioengineering
biorxiv

In vivo Calcium Imaging in the Near-Infrared II Window

XU, D.Open in Google Scholar•Wu, Z.Open in Google Scholar•Li, W. J.Open in Google Scholar•Yue, X.Open in Google Scholar•Wang, G.-L.Open in Google Scholar•Xu, S.Open in Google Scholar•Yu, H.Open in Google Scholar•Liu, Y.Open in Google Scholar•Dai, Z.Open in Google Scholar•Yu, L.-J.Open in Google Scholaret al.

Non-invasive deep-tissue calcium imaging of live mammals with high sensitivity and resolution is challenging owing to light scattering experienced by traditional calcium ion (Ca2+) indicators with excitation and emission wavelengths within 400-750 nm. Here, we report near-infrared II (NIR-II) calcium imaging beyond 1000 nm by exploring a natural protein derived from a bacterium (Thermochromatium tepidum) living in a calcium carbonate-rich environment. This highly photostable fluorescent protein enables NIR-II imaging of intracellular Ca2+ responses to stimulant drugs in cultured mammalian cells with sensitivity comparable to that of visible Ca2+ indicators. We achieve in vivo NIR-II imaging of Ca2+ transients in response to two different tumor treatment strategies in intact tumors with high sensitivity, resolution, and contrast, opening the possibility of non-invasive deep-tissue calcium imaging for assessing treatment efficacy longitudinally. This work represents the first in vivo calcium sensing in the 1000-1300 nm range of the NIR-II window.

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