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bioRxiv · 10.1101/2025.06.26.661443

In vivo Calcium Imaging in the Near-Infrared II Window

XU, D.·Wu, Z.·Li, W. J.·Yue, X.·Wang, G.-L.·Xu, S.·Yu, H.·Liu, Y.·Dai, Z.·Yu, L.-J.·Dai, H.·Wang, F.
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Abstract

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. TOC Graphic O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=105 SRC="FIGDIR/small/661443v2_ufig1.gif" ALT="Figure 1"> View larger version (24K): org.highwire.dtl.DTLVardef@149504eorg.highwire.dtl.DTLVardef@37722borg.highwire.dtl.DTLVardef@2878a7org.highwire.dtl.DTLVardef@1091d0c_HPS_FORMAT_FIGEXP M_FIG C_FIG

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XU, D., Wu, Z., Li, W. J., Yue, X., Wang, G.-L., Xu, S., Yu, H., Liu, Y., Dai, Z., Yu, L.-J., Dai, H., Wang, F.. 2025-06-28. In vivo Calcium Imaging in the Near-Infrared II Window. https://doi.org/10.1101/2025.06.26.661443

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