Light-sheet microscopy in the near-infrared II window
Open accessAbstract
Non-invasive deep-tissue three-dimensional optical imaging of live mammals with high spatiotemporal resolution is challenging owing to light scattering. We developed near-infrared II (1,000-1,700 nm) light-sheet microscopy with excitation and emission of up to approximately 1,320 nm and 1,700 nm, respectively, for optical sectioning at a penetration depth of approximately 750 μm through live tissues without invasive surgery and at a depth of approximately 2 mm in glycerol-cleared brain tissues. Near-infrared II light-sheet microscopy in normal and oblique configurations enabled in vivo imaging of live mice through intact tissue, revealing abnormal blood flow and T-cell motion in tumor microcirculation and mapping out programmed-death ligand 1 and programmed cell death protein 1 in tumors with cellular resolution. Three-dimensional imaging through the intact mouse head resolved vascular channels between the skull and brain cortex, and allowed monitoring of recruitment of macrophages and microglia to the traumatic brain injury site.
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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 [196]
The Results section reports lateral feature widths of about 7–9 μm at imaging depths of 1–2.5 mm, supporting the point for this paper’s NIR-II light-sheet microscopy. The Introduction discusses other deep-imaging techniques but does not establish that none of them has achieved sub-1-μm resolution at high depth. (LLM, conf=0.82)
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 [196]
Light-sheet microscopy with SWIR emitting inorganic nanoparticles using 1320 nm excitation light achieved the lowest degree of intensity decay and the least light-sheet thickness broadening.
“The lowest degree of intensity decay and the least light-sheet thickness broadening were achieved using this method.196”
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 [196]
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References 61 · Cited by 237
- Authors
- Wang, F., Wan, H., Ma, Z., Zhong, Y., Sun, Q., Tian, Y., Qu, L., Du, H., Zhang, M., Li, L., Ma, H., Luo, J., Liang, Y., Li, W. J., Hong, G., Liu, L., & Dai, H.
- Affiliations
- Stanford University (US)
- University of North Carolina at Chapel Hill (US)
- Wuhan University of Technology (CN)
- VA Palo Alto Health Care System (US)
- Southern University of Science and Technology (CN)
- Shenyang Institute of Automation (CN)
- Corresponding author
- Dai, Hongjie
- ORCID
- Venue
- Nature Methods
- Published
- 2019-05-13
- Volume
- 16
- Issue
- 6
- Pages
- 545-552
- ISSN
- 1548-7091 (ISSN-L)
- 1548-7105
- PMC
- PMC6579541
- OpenAlex
- W2945756399
- Cited by
- 237
- References
- 61
- Citations by year
- 2026: 25 · 2025: 33 · 2024: 32 · 2023: 29 · 2022: 36 · 2021: 37 · 2020: 36 · 2019: 8 · 2018: 1
- FWCI
- 14.14
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Nature Methods (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- 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
- light-sheet microscopy, deep tissue imaging, light scattering, in vivo imaging, mouse brain, tumor microcirculation, T cell motility, PD-L1, PD-1, traumatic brain injury, macrophage recruitment, skull imaging, vascular imaging
- MeSH terms
- Brain Injuries, Traumatic, Animals, Brain, Female, Fluorescent Dyes, Humans, Infrared Rays, Mice, Inbred BALB C, Mice, Inbred C57BL, Microscopy, Fluorescence, Tumor Cells, Cultured, Colorectal Neoplasms, Spectroscopy, Near-Infrared, Imaging, Three-Dimensional, Xenograft Model Antitumor Assays, Mice, Optical Imaging
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