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Cited by 1,326Advanced Fluorescence Microscopy TechniquesSpectroscopy Techniques in Biomedical and Chemical ResearchCell Image Analysis Techniques

The DOI matches its Crossref record; 29 of the 29 references it cites were found in a registry or an index.

Second-harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms

Campagnola, Paul J, Loew, Leslie M·2003·Nature Biotechnology
DOI 10.1038/nbt894
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Abstract

Although the nonlinear optical effect known as second-harmonic generation (SHG) has been recognized since the earliest days of laser physics and was demonstrated through a microscope over 25 years ago, only in the past few years has it begun to emerge as a viable microscope imaging contrast mechanism for visualization of cell and tissue structure and function. Only small modifications are required to equip a standard laser-scanning two-photon microscope for second-harmonic imaging microscopy (SHIM). Recent studies of the three-dimensional in vivo structures of well-ordered protein assemblies, such as collagen, microtubules and muscle myosin, are beginning to establish SHIM as a nondestructive imaging modality that holds promise for both basic research and clinical pathology. Thus far the best signals have been obtained in a transmitted light geometry that precludes in vivo measurements on large living animals. This drawback may be addressed through improvements in the collection of SHG signals via an epi-illumination microscope configuration. In addition, SHG signals from certain membrane-bound dyes have been shown to be highly sensitive to membrane potential. Although this indicates that SHIM may become a valuable tool for probing cell physiology, the small signal size would limit the number of photons that could be collected during the course of a fast action potential. Better dyes and optimized microscope optics could ultimately lead to the imaging of neuronal electrical a…

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Research works printed but not linked: 3

  1. Sheppard, C.J.R., Kompfner, R., Gannaway, J. & Walsh, D. Scanning harmonic optical microscope. IEEE J. Quantum Electron. 13E, 100D (1977).
  2. Brown, E. et al. Dynamic imaging of collagen and its modulation in tumors in vivo using second harmonic generation. Nat. Med. 9, 796–800 (2003). · DOI
  3. Huang, J.Y., Lewis, A. & Loew, L.M. Non-linear optical properties of potential sensitive styryl dyes. Biophys. J. 53, 665–670 (1988). · DOI

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Metadata

Authors
Campagnola, P. J., & Loew, L. M.
Affiliations
UConn Health (US)
Corresponding author
Loew, Leslie M
ORCID
Venue
Nature Biotechnology
Publisher
Springer Science and Business Media LLC
Published
2003-10-31
Volume
21
Issue
11
Pages
1356-1360
ISSN
  • 0733-222X (ISSN-L)
  • 1087-0156
  • 1546-1696
DOI
10.1038/nbt894
OpenAlex
W2077572701
Cited by
1,326
References
29
Citations by year
2026: 37 · 2025: 42 · 2024: 43 · 2023: 50 · 2022: 55 · 2021: 55 · 2020: 67 · 2019: 79 · 2018: 55 · 2017: 67 · 2016: 73 · 2015: 79 · 2014: 61 · 2013: 77 · 2012: 72
FWCI
3.10
Type
journal article
Language
en
Open access
closed
Licence
www.springer.com/tdm (tdm)
Locations
  • Nature Biotechnology (publishedVersion)
  • PubMed (publishedVersion)
Funders
  • National Institutes of Health (5r21rr013472-02)
  • Office of Naval Research
  • National Institute of Biomedical Imaging and Bioengineering (R01EB00196)
  • National Center for Research Resources (R21 RR13472)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Spectroscopy Techniques in Biomedical and Chemical Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Cell Image Analysis Techniques
Keywords
second harmonic generation, nonlinear optical microscopy, collagen imaging, microtubules, myosin, membrane potential, neuronal electrical activity, in vivo imaging
MeSH terms
Action Potentials, Cell Membrane, Cells, Image Enhancement, Membrane Potentials, Proteins, Microscopy, Confocal

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