Noninvasive corneal stromal collagen imaging using two-photon-generated second-harmonic signals

Naoyuki Morishige, W. Matthew Petroll, Teruo Nishida, M. Cristina Kenney, James V. Jester

Research output: Contribution to journalArticle

115 Citations (Scopus)

Abstract

Purpose: To investigate the feasibility of using femtosecond-pulse lasers to produce second-harmonic generated (SHG) signals to noninvasively assess corneal stromal collagen organization. Setting: The Eye Institute, University of California, Irvine, California, USA. Methods: Mouse, rabbit, and human corneas were examined by two-photon confocal microscopy using a variable-wavelength femtosecond lasers to produce SHG signals. Two types were detected: forward scattered and backward scattered. Wavelength dependence of the SHG signal was confirmed by spectral separation using the 510 Meta (Zeiss). To verify the spatial relation between SHG signals and corneal cells, staining of cytoskeletons and nuclei was performed. Results: Second-harmonic-generated signal intensity was strongest with an excitation wavelength of 800 nm for all 3 species. Second-harmonic-generated forward signals showed a distinct fibrillar pattern organized into bands suggesting lamellae, while backscattered SHG signals appeared more diffuse and indistinct. Reconstruction of SHG signals showed two patterns of lamellar organization: highly interwoven in the anterior stroma and orthogonally arranged in the posterior stroma. Unique to the human cornea was the presence of transverse, sutural lamellae that inserted into Bowman's layer, suggesting an anchoring function. Conclusions: Using two-photon confocal microscopy to generate SHG signals from the corneal collagen provides a powerful new approach to noninvasively study corneal structure. Human corneas had a unique organizational pattern with sutural lamellae to provide important biomechanical support that was not present in mouse or rabbit corneas.

Original languageEnglish (US)
Pages (from-to)1784-1791
Number of pages8
JournalJournal of Cataract and Refractive Surgery
Volume32
Issue number11
DOIs
StatePublished - Nov 2006

Fingerprint

Photons
Cornea
Collagen
Confocal Microscopy
Lasers
Organizations
Rabbits
Cytoskeleton
Staining and Labeling

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Noninvasive corneal stromal collagen imaging using two-photon-generated second-harmonic signals. / Morishige, Naoyuki; Petroll, W. Matthew; Nishida, Teruo; Kenney, M. Cristina; Jester, James V.

In: Journal of Cataract and Refractive Surgery, Vol. 32, No. 11, 11.2006, p. 1784-1791.

Research output: Contribution to journalArticle

Morishige, Naoyuki ; Petroll, W. Matthew ; Nishida, Teruo ; Kenney, M. Cristina ; Jester, James V. / Noninvasive corneal stromal collagen imaging using two-photon-generated second-harmonic signals. In: Journal of Cataract and Refractive Surgery. 2006 ; Vol. 32, No. 11. pp. 1784-1791.
@article{7916953060bb46489b51faffe2a72dc6,
title = "Noninvasive corneal stromal collagen imaging using two-photon-generated second-harmonic signals",
abstract = "Purpose: To investigate the feasibility of using femtosecond-pulse lasers to produce second-harmonic generated (SHG) signals to noninvasively assess corneal stromal collagen organization. Setting: The Eye Institute, University of California, Irvine, California, USA. Methods: Mouse, rabbit, and human corneas were examined by two-photon confocal microscopy using a variable-wavelength femtosecond lasers to produce SHG signals. Two types were detected: forward scattered and backward scattered. Wavelength dependence of the SHG signal was confirmed by spectral separation using the 510 Meta (Zeiss). To verify the spatial relation between SHG signals and corneal cells, staining of cytoskeletons and nuclei was performed. Results: Second-harmonic-generated signal intensity was strongest with an excitation wavelength of 800 nm for all 3 species. Second-harmonic-generated forward signals showed a distinct fibrillar pattern organized into bands suggesting lamellae, while backscattered SHG signals appeared more diffuse and indistinct. Reconstruction of SHG signals showed two patterns of lamellar organization: highly interwoven in the anterior stroma and orthogonally arranged in the posterior stroma. Unique to the human cornea was the presence of transverse, sutural lamellae that inserted into Bowman's layer, suggesting an anchoring function. Conclusions: Using two-photon confocal microscopy to generate SHG signals from the corneal collagen provides a powerful new approach to noninvasively study corneal structure. Human corneas had a unique organizational pattern with sutural lamellae to provide important biomechanical support that was not present in mouse or rabbit corneas.",
author = "Naoyuki Morishige and Petroll, {W. Matthew} and Teruo Nishida and Kenney, {M. Cristina} and Jester, {James V.}",
year = "2006",
month = "11",
doi = "10.1016/j.jcrs.2006.08.027",
language = "English (US)",
volume = "32",
pages = "1784--1791",
journal = "Journal of Cataract and Refractive Surgery",
issn = "0886-3350",
publisher = "Elsevier Inc.",
number = "11",

}

TY - JOUR

T1 - Noninvasive corneal stromal collagen imaging using two-photon-generated second-harmonic signals

AU - Morishige, Naoyuki

AU - Petroll, W. Matthew

AU - Nishida, Teruo

AU - Kenney, M. Cristina

AU - Jester, James V.

PY - 2006/11

Y1 - 2006/11

N2 - Purpose: To investigate the feasibility of using femtosecond-pulse lasers to produce second-harmonic generated (SHG) signals to noninvasively assess corneal stromal collagen organization. Setting: The Eye Institute, University of California, Irvine, California, USA. Methods: Mouse, rabbit, and human corneas were examined by two-photon confocal microscopy using a variable-wavelength femtosecond lasers to produce SHG signals. Two types were detected: forward scattered and backward scattered. Wavelength dependence of the SHG signal was confirmed by spectral separation using the 510 Meta (Zeiss). To verify the spatial relation between SHG signals and corneal cells, staining of cytoskeletons and nuclei was performed. Results: Second-harmonic-generated signal intensity was strongest with an excitation wavelength of 800 nm for all 3 species. Second-harmonic-generated forward signals showed a distinct fibrillar pattern organized into bands suggesting lamellae, while backscattered SHG signals appeared more diffuse and indistinct. Reconstruction of SHG signals showed two patterns of lamellar organization: highly interwoven in the anterior stroma and orthogonally arranged in the posterior stroma. Unique to the human cornea was the presence of transverse, sutural lamellae that inserted into Bowman's layer, suggesting an anchoring function. Conclusions: Using two-photon confocal microscopy to generate SHG signals from the corneal collagen provides a powerful new approach to noninvasively study corneal structure. Human corneas had a unique organizational pattern with sutural lamellae to provide important biomechanical support that was not present in mouse or rabbit corneas.

AB - Purpose: To investigate the feasibility of using femtosecond-pulse lasers to produce second-harmonic generated (SHG) signals to noninvasively assess corneal stromal collagen organization. Setting: The Eye Institute, University of California, Irvine, California, USA. Methods: Mouse, rabbit, and human corneas were examined by two-photon confocal microscopy using a variable-wavelength femtosecond lasers to produce SHG signals. Two types were detected: forward scattered and backward scattered. Wavelength dependence of the SHG signal was confirmed by spectral separation using the 510 Meta (Zeiss). To verify the spatial relation between SHG signals and corneal cells, staining of cytoskeletons and nuclei was performed. Results: Second-harmonic-generated signal intensity was strongest with an excitation wavelength of 800 nm for all 3 species. Second-harmonic-generated forward signals showed a distinct fibrillar pattern organized into bands suggesting lamellae, while backscattered SHG signals appeared more diffuse and indistinct. Reconstruction of SHG signals showed two patterns of lamellar organization: highly interwoven in the anterior stroma and orthogonally arranged in the posterior stroma. Unique to the human cornea was the presence of transverse, sutural lamellae that inserted into Bowman's layer, suggesting an anchoring function. Conclusions: Using two-photon confocal microscopy to generate SHG signals from the corneal collagen provides a powerful new approach to noninvasively study corneal structure. Human corneas had a unique organizational pattern with sutural lamellae to provide important biomechanical support that was not present in mouse or rabbit corneas.

UR - http://www.scopus.com/inward/record.url?scp=33751043199&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33751043199&partnerID=8YFLogxK

U2 - 10.1016/j.jcrs.2006.08.027

DO - 10.1016/j.jcrs.2006.08.027

M3 - Article

C2 - 17081858

AN - SCOPUS:33751043199

VL - 32

SP - 1784

EP - 1791

JO - Journal of Cataract and Refractive Surgery

JF - Journal of Cataract and Refractive Surgery

SN - 0886-3350

IS - 11

ER -