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
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…
Overview
Integrity signals
Registry
Cites
Statistics
Verification
Claims the paper makes
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
In the citing paper
“second harmonic generation (SHG) microscopy has emerged as a label-free imaging modality capable of visualizing collagen-rich structures within intact spheroids [259,260].”
From Modeling Tumor Microenvironment Complexity In Vitro: Spheroids as Physiologically Relevant Tumor Models and Strategies for Their Analysis (2025) · cites this paper as [260]
Citations
Cites
Zipfel, W. R., Williams, R. M., Christie, R. H., Nikitin, A. Y., Hyman, B. T., & Webb, W. W. (2003) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0832308100 · Cited by 1,712
Campagnola, P. J., Millard, A. C., Terasaki, M., Hoppe, P. E., Malone, C. J., & Mohler, W. A. (2002) · Biophysical Journal · doi:10.1016/s0006-3495(02)75414-3 · Cited by 922
Self-citationZoumi, A., Yeh, A. T., & Tromberg, B. J. (2002) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.172368799 · Cited by 872
Dynamic imaging of collagen and its modulation in tumors in vivo using second-harmonic generation
Brown, E. B., McKee, T. D., DiTomaso, E., Pluen, A., Seed, B., Boucher, Y., & Jain, R. K. (2003) · Nature Medicine · doi:10.1038/nm879 · Cited by 844
High-Resolution Nonlinear Optical Imaging of Live Cells by Second Harmonic Generation
Campagnola, P. J., Wei, M.-D., Lewis, A., & Loew, L. M. (1999) · Biophysical Journal · doi:10.1016/s0006-3495(99)77165-1 · Cited by 504
Self-citationFreund, I., Deutsch, M., & Sprecher, A. (1986) · Biophysical Journal · doi:10.1016/s0006-3495(86)83510-x · Cited by 449
3-Dimensional imaging of collagen using second harmonic generation
Cox, G., Kable, E., Jones, A., Fraser, I., Manconi, F., & Gorrell, M. D. (2003) · Journal of Structural Biology · doi:10.1016/s1047-8477(02)00576-2 · Cited by 430
Polarization-Modulated Second Harmonic Generation in Collagen
Stoller, P. C., Reiser, K. M., Celliers, P. M., & Rubenchik, A. M. (2002) · Biophysical Journal · doi:10.1016/s0006-3495(02)75673-7 · Cited by 403
Membrane imaging by second-harmonic generation microscopy
Moreaux, L. C., Sandre, O., & Mertz, J. C. (2000) · Journal of the Optical Society of America B · doi:10.1364/josab.17.001685 · Cited by 346
Inoué, S. (1986) · Springer eBooks · doi:10.1007/978-1-4757-6925-8 · Cited by 343
Dombeck, D. A., Kasischke, K. A., Vishwasrao, H. D., Ingelsson, M., Hyman, B. T., & Webb, W. W. (2003) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0731953100 · Cited by 263
Second harmonic generation imaging of endogenous structural proteins
Mohler, W. A., Millard, A. C., & Campagnola, P. J. (2003) · Methods · doi:10.1016/s1046-2023(02)00292-x · Cited by 259
Self-citationSecond harmonic imaging microscopy of living cells
Campagnola, P. J., Clark, H. A., Mohler, W. A., Lewis, A., & Loew, L. M. (2001) · Journal of Biomedical Optics · doi:10.1117/1.1383294 · Cited by 245
Self-citationPolarization-dependent optical second-harmonic imaging of rat-tail tendon
Stoller, P. C., Kim, B., Rubenchik, A. M., Reiser, K. M., & da Silva, L. B. (2002) · Journal of Biomedical Optics · doi:10.1117/1.1431967 · Cited by 224
Coherent Scattering in Multi-Harmonic Light Microscopy
Moreaux, L. C., Sandre, O., Charpak, S., Blanchard-Desce, M. H., & Mertz, J. C. (2001) · Biophysical Journal · doi:10.1016/s0006-3495(01)76129-2 · Cited by 222
Membrane imaging by simultaneous second-harmonic generation and two-photon microscopy
Moreaux, L. C., Sandre, O., Blanchard-Desce, M. H., & Mertz, J. C. (2000) · Optics Letters · doi:10.1364/ol.25.000320 · Cited by 209
Nonlinear optical microscopic examination of structure in polycrystalline ZnSe
Hellwarth, R. W., & Christensen, P. (1974) · Optics Communications · doi:10.1016/0030-4018(74)90024-8 · Cited by 175
Yeh, A. T., Nassif, N. A., Zoumi, A., & Tromberg, B. J. (2002) · Optics Letters · doi:10.1364/ol.27.002082 · Cited by 170
Probing membrane potential with nonlinear optics
Bouevitch, O., Lewis, A., Pinevsky, I., Wuskell, J. P., & Loew, L. M. (1993) · Biophysical Journal · doi:10.1016/s0006-3495(93)81126-3 · Cited by 139
Self-citationPeleg, G., Lewis, A., Michal Linial, B., & Loew, L. M. (1999) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.96.12.6700 · Cited by 123
Self-citationElectro-optic response of second-harmonic generation membrane potential sensors
Moreaux, L. C., Pons, T., Dambrin, V., Blanchard-Desce, M. H., & Mertz, J. C. (2003) · Optics Letters · doi:10.1364/ol.28.000625 · Cited by 68
Second Harmonic Generation Properties of Fluorescent Polymer-Encapsulated Gold Nanoparticles
Clark, H. A., Campagnola, P. J., Wuskell, J. P., Lewis, A., & Loew, L. M. (2000) · Journal of the American Chemical Society · doi:10.1021/ja002223v · Cited by 67
Self-citationMillard, A. C., Jin, L., Lewis, A., & Loew, L. M. (2003) · Optics Letters · doi:10.1364/ol.28.001221 · Cited by 58
Self-citation[3] Second harmonic imaging microscopy
Millard, A. C., Campagnola, P. J., Mohler, W. A., Lewis, A., & Loew, L. M. (2003) · Methods in enzymology on CD-ROM/Methods in enzymology · doi:10.1016/s0076-6879(03)61005-0 · Cited by 57
Self-citationNonlinear optical properties of potential sensitive styryl dyes
Huang, J. Y., Lewis, A., & Loew, L. M. (1988) · Biophysical Journal · doi:10.1016/s0006-3495(88)83147-3 · Cited by 57
Self-citationInfrared nonlinear optical measurements of membrane potential in photoreceptor cells
Ben-Oren, I., Peleg, G., Lewis, A., Minke, B., & Loew, L. M. (1996) · Biophysical Journal · doi:10.1016/s0006-3495(96)79365-7 · Cited by 56
Self-citationIn situdetermination of induced dipole moments of pure and membrane-bound retinal chromophores
Rasing, T., Huang, J., Lewis, A., Stehlin, T., & Shen, Y. R. (1989) · Physical Review A · doi:10.1103/physreva.40.1684 · Cited by 26
Image formation in three-photon fluorescence microscopy
Sheppard, C. J. R. (1996) · Bioimaging · doi:10.1002/1361-6374(199609)4:3 · Cited by 20
Self-citationPotentiometric Membrane Dyes and Imaging Membrane Potential in Single Cells
Loew, L. M. (1999) · Elsevier eBooks · doi:10.1016/b978-012447836-7/50016-6 · Cited by 11
Self-citation
Flagged first, then by citations
Research works printed but not linked: 3
- Sheppard, C.J.R., Kompfner, R., Gannaway, J. & Walsh, D. Scanning harmonic optical microscope. IEEE J. Quantum Electron. 13E, 100D (1977).
- 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
- Huang, J.Y., Lewis, A. & Loew, L.M. Non-linear optical properties of potential sensitive styryl dyes. Biophys. J. 53, 665–670 (1988). · DOI
Cited by
Sorted by citations
Deep tissue two-photon microscopy
Helmchen, F., & Denk, W. T. (2005) · Nature Methods · doi:10.1038/nmeth818 · Cited by 4,288
Upconversion Nanoparticles: Design, Nanochemistry, and Applications in Theranostics
Chen, G., Qiu, H., Prasad, P. N., & Chen, X. S. (2014) · Chemical Reviews · doi:10.1021/cr400425h · Cited by 2,746
Fluorescence Lifetime Measurements and Biological Imaging
Berezin, M. Y., & Achilefu, S. (2010) · Chemical Reviews · doi:10.1021/cr900343z · Cited by 2,463
Going deeper than microscopy: the optical imaging frontier in biology
Ntziachristos, V. (2010) · Nature Methods · doi:10.1038/nmeth.1483 · Cited by 1,896
Nanochemistry and Nanomedicine for Nanoparticle-based Diagnostics and Therapy
Chen, G., Roy, I., Yang, C., & Prasad, P. N. (2016) · Chemical Reviews · doi:10.1021/acs.chemrev.5b00148 · Cited by 1,528
Metadata
- Authors
- Campagnola, P. J., & Loew, L. M.
- Affiliations
- UConn Health (US)
- Corresponding author
- Loew, Leslie M
- Venue
- Nature Biotechnology
- Published
- 2003-10-31
- Volume
- 21
- Issue
- 11
- Pages
- 1356-1360
- ISSN
- 0733-222X (ISSN-L)
- 1087-0156
- 1546-1696
- 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
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
