In vivo NIR-II structured-illumination light-sheet microscopy
Open accessAbstract
Significance: Three-dimensional microscopy in the NIR-II window (1,000 to 1,700 nm) allows noninvasive deep-tissue optical sectioning of live mammals with high spatiotemporal resolution due to suppressed light scattering and reduced tissue autofluorescence. Herein, we present a NIR-II structured-illumination light-sheet microscopy (NIR-II SIM) with both excitation and emission wavelengths in the NIR-IIb window (1,500 to 1,700 nm). Integrating structured illumination into NIR-II light-sheet microscopy further diminished out-of-focus background and improved spatial resolution, facilitating in vivo imaging of cell behavior in the intact tissues without any invasive surgery during a long period. NIR-II SIM enabled longitudinally multiplexed molecular imaging of immune cells in response to immunotherapy in the tumor microenvironment.
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A spatial resolution below 1 μm at a high depth has not been achieved by these imaging techniques.
“However, a spatial resolution below 1 μm at a high depth has not been achieved by these imaging techniques.”
From The Pursuit of Shortwave Infrared-Emitting Nanoparticles with Bright Fluorescence through Molecular Design and Excited-State Engineering of Molecular Aggregates (2022) · cites this paper as [197]
The Results report a vascular feature width of 9 μm at approximately 1 mm depth, consistent with a resolution–depth trade-off in that experiment. But the Discussion also reports approximately 0.96 μm lateral resolution for NIR-II SIM using a high-NA objective; the paper does not establish the categorical claim that none of the imaging techniques achieved sub-1-μm resolution at high depth. (LLM, conf=0.78)
Experimental — accuracy not yet measuredA penetration depth up to ∼750 μm was achieved in through-scalp and -skull imaging of intact mouse head without craniotomy and through-skin imaging of tumors.
“A penetration depth up to ∼750 μm was achieved in the through-scalp and -skull imaging of intact mouse head without craniotomy196 and through-skin imaging of tumors.197”
From The Pursuit of Shortwave Infrared-Emitting Nanoparticles with Bright Fluorescence through Molecular Design and Excited-State Engineering of Molecular Aggregates (2022) · cites this paper as [197]
Integration of structured illumination microscopy with SWIR light-sheet microscopy with ultralong excitation and emission wavelengths up to ∼1540 and ∼1700 nm led to further diminished background and improved spatial resolution by approximately twofold.
“An integration of structured illumination microscopy with the SWIR light-sheet microscopy with ultralong excitation and emission wavelengths up to ∼1540 and ∼1700 nm, respectively, lead to further diminished background and improved spatial resolution by approximately twofold.197”
From The Pursuit of Shortwave Infrared-Emitting Nanoparticles with Bright Fluorescence through Molecular Design and Excited-State Engineering of Molecular Aggregates (2022) · cites this paper as [197]
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- Authors
- Wang, F., Ma, Z., Zhong, Y., Salazar, F., Xu, C., Ren, F., Qu, L., Wu, A. M., & Dai, H.
- Affiliations
- Stanford University (US)
- City Of Hope National Medical Center (US)
- Corresponding author
- Wang, Feifei
- Ma, Zhuoran
- Dai, Hongjie
- ORCID
- Venue
- Proceedings of the National Academy of Sciences
- Publisher
- National Academy of Sciences
- Published
- 2021-02
- Volume
- 118
- Issue
- 6
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- PMC
- PMC8017937
- OpenAlex
- W3127300648
- Cited by
- 85
- References
- 79
- Citations by year
- 2026: 11 · 2025: 17 · 2024: 17 · 2023: 17 · 2022: 18 · 2021: 5
- FWCI
- 6.46
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Proceedings of the National Academy of Sciences (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institute of Neurological Disorders and Stroke (DP1NS105737)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
- Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Optical Imaging and Spectroscopy Techniques
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Spectroscopy Techniques in Biomedical and Chemical Research
- Keywords
- structured illumination, in vivo imaging, deep tissue imaging, light scattering suppression, tumor microenvironment, CT26 cells, OX40, CD4 and CD8 T cells, immunotherapy, CpG, Toll-like receptor 9 agonist
- MeSH terms
- Animals, Microscopy, Fluorescence, Imaging, Three-Dimensional, Cell Line, Tumor, Toll-Like Receptor 9, Mice, Single-Cell Analysis, Cellular Microenvironment, Optical Imaging
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