Quantitative analysis of collagen lamellae in the normal and keratoconic human cornea by second harmonic generation imaging microscopy

Naoyuki Morishige, Ryutaro Shin-Gyou-Uchi, Haruya Azumi, Hiroaki Ohta, Yukiko Morita, Naoyuki Yamada, Kazuhiro Kimura, Atsushi Takahara, Koh Hei Sonoda

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Abstract

Purpose. To characterize the structural properties of collagen lamellae in the normal and keratoconic human corneal stroma, we measured their width and angle relative to Bowman's layer (BL).

Methods. Thirteen normal and four keratoconic corneas were examined. Collagen lamellae in tissue blocks from the central cornea were visualized by second harmonic generation imaging microscopy. Images obtained in 1-μm steps from BL to Descemet's membrane (DM) were subjected to three-dimensional reconstruction. The reconstructed data sets were divided into 10 layers of equal depth (L1–L10) for analysis. The width of lamellae adherent to BL (L0) was also determined.

Results. For the normal cornea, the width (mean ± SD) of collagen lamellae was 6.5 ± 1.7 μm at L0, decreased to 4.3 ± 1.3 μm at L1, and then increased gradually with progression toward DM to 122.2 ± 34.5 μm at L10, whereas the angle of lamellae was 20.9° ± 5.4° at L1 and decreased initially to 10.6° ± 3.2° at L2 before declining gradually to 2.7° ± 2.2° at L10. The width and angle of collagen lamellae in the keratoconic cornea were significantly larger and smaller, respectively, relative to those in the normal cornea.

Conclusions. In the normal human cornea, collagen lamellae adjacent to BL are narrow and form a steep angle with BL, whereas they increase in width and their angle relative to BL flattens with progression toward DM. These properties of collagen lamellae are altered in keratoconus and are likely related to abnormalities of corneal shape.

Original languageEnglish
Pages (from-to)8377-8385
Number of pages9
JournalInvestigative Ophthalmology and Visual Science
Volume55
Issue number12
DOIs
Publication statusPublished - Dec 2014

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Cornea
Microscopy
Collagen
Descemet Membrane
Corneal Stroma
Keratoconus

All Science Journal Classification (ASJC) codes

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

Quantitative analysis of collagen lamellae in the normal and keratoconic human cornea by second harmonic generation imaging microscopy. / Morishige, Naoyuki; Shin-Gyou-Uchi, Ryutaro; Azumi, Haruya; Ohta, Hiroaki; Morita, Yukiko; Yamada, Naoyuki; Kimura, Kazuhiro; Takahara, Atsushi; Sonoda, Koh Hei.

In: Investigative Ophthalmology and Visual Science, Vol. 55, No. 12, 12.2014, p. 8377-8385.

Research output: Contribution to journalArticle

Morishige, Naoyuki ; Shin-Gyou-Uchi, Ryutaro ; Azumi, Haruya ; Ohta, Hiroaki ; Morita, Yukiko ; Yamada, Naoyuki ; Kimura, Kazuhiro ; Takahara, Atsushi ; Sonoda, Koh Hei. / Quantitative analysis of collagen lamellae in the normal and keratoconic human cornea by second harmonic generation imaging microscopy. In: Investigative Ophthalmology and Visual Science. 2014 ; Vol. 55, No. 12. pp. 8377-8385.
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abstract = "Purpose. To characterize the structural properties of collagen lamellae in the normal and keratoconic human corneal stroma, we measured their width and angle relative to Bowman's layer (BL).Methods. Thirteen normal and four keratoconic corneas were examined. Collagen lamellae in tissue blocks from the central cornea were visualized by second harmonic generation imaging microscopy. Images obtained in 1-μm steps from BL to Descemet's membrane (DM) were subjected to three-dimensional reconstruction. The reconstructed data sets were divided into 10 layers of equal depth (L1–L10) for analysis. The width of lamellae adherent to BL (L0) was also determined.Results. For the normal cornea, the width (mean ± SD) of collagen lamellae was 6.5 ± 1.7 μm at L0, decreased to 4.3 ± 1.3 μm at L1, and then increased gradually with progression toward DM to 122.2 ± 34.5 μm at L10, whereas the angle of lamellae was 20.9° ± 5.4° at L1 and decreased initially to 10.6° ± 3.2° at L2 before declining gradually to 2.7° ± 2.2° at L10. The width and angle of collagen lamellae in the keratoconic cornea were significantly larger and smaller, respectively, relative to those in the normal cornea.Conclusions. In the normal human cornea, collagen lamellae adjacent to BL are narrow and form a steep angle with BL, whereas they increase in width and their angle relative to BL flattens with progression toward DM. These properties of collagen lamellae are altered in keratoconus and are likely related to abnormalities of corneal shape.",
author = "Naoyuki Morishige and Ryutaro Shin-Gyou-Uchi and Haruya Azumi and Hiroaki Ohta and Yukiko Morita and Naoyuki Yamada and Kazuhiro Kimura and Atsushi Takahara and Sonoda, {Koh Hei}",
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T1 - Quantitative analysis of collagen lamellae in the normal and keratoconic human cornea by second harmonic generation imaging microscopy

AU - Morishige, Naoyuki

AU - Shin-Gyou-Uchi, Ryutaro

AU - Azumi, Haruya

AU - Ohta, Hiroaki

AU - Morita, Yukiko

AU - Yamada, Naoyuki

AU - Kimura, Kazuhiro

AU - Takahara, Atsushi

AU - Sonoda, Koh Hei

PY - 2014/12

Y1 - 2014/12

N2 - Purpose. To characterize the structural properties of collagen lamellae in the normal and keratoconic human corneal stroma, we measured their width and angle relative to Bowman's layer (BL).Methods. Thirteen normal and four keratoconic corneas were examined. Collagen lamellae in tissue blocks from the central cornea were visualized by second harmonic generation imaging microscopy. Images obtained in 1-μm steps from BL to Descemet's membrane (DM) were subjected to three-dimensional reconstruction. The reconstructed data sets were divided into 10 layers of equal depth (L1–L10) for analysis. The width of lamellae adherent to BL (L0) was also determined.Results. For the normal cornea, the width (mean ± SD) of collagen lamellae was 6.5 ± 1.7 μm at L0, decreased to 4.3 ± 1.3 μm at L1, and then increased gradually with progression toward DM to 122.2 ± 34.5 μm at L10, whereas the angle of lamellae was 20.9° ± 5.4° at L1 and decreased initially to 10.6° ± 3.2° at L2 before declining gradually to 2.7° ± 2.2° at L10. The width and angle of collagen lamellae in the keratoconic cornea were significantly larger and smaller, respectively, relative to those in the normal cornea.Conclusions. In the normal human cornea, collagen lamellae adjacent to BL are narrow and form a steep angle with BL, whereas they increase in width and their angle relative to BL flattens with progression toward DM. These properties of collagen lamellae are altered in keratoconus and are likely related to abnormalities of corneal shape.

AB - Purpose. To characterize the structural properties of collagen lamellae in the normal and keratoconic human corneal stroma, we measured their width and angle relative to Bowman's layer (BL).Methods. Thirteen normal and four keratoconic corneas were examined. Collagen lamellae in tissue blocks from the central cornea were visualized by second harmonic generation imaging microscopy. Images obtained in 1-μm steps from BL to Descemet's membrane (DM) were subjected to three-dimensional reconstruction. The reconstructed data sets were divided into 10 layers of equal depth (L1–L10) for analysis. The width of lamellae adherent to BL (L0) was also determined.Results. For the normal cornea, the width (mean ± SD) of collagen lamellae was 6.5 ± 1.7 μm at L0, decreased to 4.3 ± 1.3 μm at L1, and then increased gradually with progression toward DM to 122.2 ± 34.5 μm at L10, whereas the angle of lamellae was 20.9° ± 5.4° at L1 and decreased initially to 10.6° ± 3.2° at L2 before declining gradually to 2.7° ± 2.2° at L10. The width and angle of collagen lamellae in the keratoconic cornea were significantly larger and smaller, respectively, relative to those in the normal cornea.Conclusions. In the normal human cornea, collagen lamellae adjacent to BL are narrow and form a steep angle with BL, whereas they increase in width and their angle relative to BL flattens with progression toward DM. These properties of collagen lamellae are altered in keratoconus and are likely related to abnormalities of corneal shape.

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