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Selective corneal imaging using combined second-harmonic generation and two-photon excited fluorescence
Open accessAbstract
A multiphoton microscope employing second-harmonic generation (SHG) and two-photon excited fluorescence (TPF) is used for high-resolution ex vivo imaging of rabbit cornea in a backscattering geometry. Endogenous TPF and SHG signals from corneal cells and extracellular matrix, respectively, are clearly visible without exogenous dyes. Spectral characterization of these upconverted signals provides confirmation of the structural origin of both TPF and SHG, and spectral imaging facilitates the separation of keratocyte and epithelial cells from the collagen-rich corneal stroma. The polarization dependence of collagen SHG is used to highlight fiber orientation, and three-dimensional SHG tomography reveals that approximately 88% of the stromal volume is occupied by collagen lamellae.
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Citations
Cited by
Campagnola, P. J., & Loew, L. M. (2003) · Nature Biotechnology · doi:10.1038/nbt894 · Cited by 1,326
Second harmonic generation microscopy for quantitative analysis of collagen fibrillar structure
Chen, X., Nadiarynkh, O., Plotnikov, S., & Campagnola, P. J. (2012) · Nature Protocols · doi:10.1038/nprot.2012.009 · Cited by 985
Characterization of the Myosin-Based Source for Second-Harmonic Generation from Muscle Sarcomeres
Plotnikov, S. V., Millard, A. C., Campagnola, P. J., & Mohler, W. A. (2006) · Biophysical Journal · doi:10.1529/biophysj.105.071555 · Cited by 444
Single Cell Optical Imaging and Spectroscopy
Stender, A. S., Marchuk, K., Liu, C., Sander, S., Meyer, M. W., Smith, E. A., Neupane, B., Wang, G., Li, J., Cheng, J.-X., et al. (2013) · Chemical Reviews · doi:10.1021/cr300336e · Cited by 292
Noninvasive corneal stromal collagen imaging using two-photon-generated second-harmonic signals
Morishige, N., Petroll, M. W., Nishida, T., Kenney, C. M., & Jester, J. V. (2006) · Journal of Cataract and Refractive Surgery · doi:10.1016/j.jcrs.2006.08.027 · Cited by 154
Metadata
- Authors
- Yeh, A. T., Nassif, N., Zoumi, A., & Tromberg, B. J.
- Venue
- Optics Letters
- Publisher
- Optica Publishing Group
- Published
- 2002-12-01
- Volume
- 27
- Issue
- 23
- Pages
- 2082
- ISSN
- 0146-9592 (ISSN-L)
- 1539-4794
- OpenAlex
- W2067634317
- Cited by
- 170
- References
- 0
- FWCI
- 3.83
- Type
- journal article
- Language
- en
- Open access
- green
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- Yeh, A. T., Nassif, N., Zoumi, A., & Tromberg, B. J. (2002). Selective corneal imaging using combined second-harmonic generation and two-photon excited fluorescence. Optics Letters. https://doi.org/10.1364/ol.27.002082
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