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Related Experiment Video

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In Vivo 2-Photon Calcium Imaging in Layer 2/3 of Mice
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In Vivo Calcium Imaging in the Near-Infrared II Window.

Danyang Xu1,2, Zhisheng Wu1, Hui Yang3

  • 1Department of Electrical and Electronic Engineering, The University of Hong Kong, Kowloon, Hong Kong SAR 999077, China.

Journal of the American Chemical Society
|December 24, 2025
PubMed
Summary

Researchers developed a novel near-infrared II (NIR-II) calcium imaging technique using a bacterial protein. This method allows for high-sensitivity, deep-tissue imaging of calcium ion (Ca2+) dynamics in live mammals, aiding in tumor treatment assessment.

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Area of Science:

  • Biomedical Optics
  • Molecular Imaging
  • Biotechnology

Background:

  • Traditional calcium ion (Ca2+) imaging in deep tissues of live mammals is limited by light scattering at visible wavelengths (400-750 nm).
  • High sensitivity and resolution are crucial for noninvasive imaging, especially for monitoring dynamic biological processes and treatment responses.

Purpose of the Study:

  • To develop a novel near-infrared II (NIR-II) window calcium imaging technique for noninvasive deep-tissue imaging in live mammals.
  • To explore a natural protein from *Thermochromatium tepidum* for enhanced Ca2+ indicator properties in the NIR-II spectrum (>1000 nm).
  • To demonstrate the utility of this NIR-II Ca2+ imaging for assessing tumor treatment efficacy longitudinally.

Main Methods:

  • Exploration and characterization of a novel, highly photostable fluorescent protein derived from *Thermochromatium tepidum*.
  • Utilized the protein for intracellular Ca2+ imaging in cultured mammalian cells, comparing sensitivity with visible Ca2+ indicators.
  • Performed *in vivo* NIR-II Ca2+ imaging in intact tumors in live mammals to monitor responses to different treatment strategies.

Main Results:

  • The developed bacterial protein enables sensitive NIR-II Ca2+ imaging of intracellular Ca2+ responses in mammalian cells.
  • Achieved high-sensitivity, high-resolution, and high-contrast *in vivo* imaging of Ca2+ transients in tumors using NIR-II wavelengths.
  • Demonstrated the potential for longitudinal assessment of tumor treatment efficacy through noninvasive deep-tissue Ca2+ imaging.

Conclusions:

  • A novel NIR-II fluorescent protein enables advanced deep-tissue calcium imaging beyond 1000 nm.
  • This technology offers a promising avenue for noninvasive monitoring of biological processes and therapeutic responses in live mammals.
  • Opens possibilities for longitudinal evaluation of treatment efficacy in various biomedical applications.