Second-harmonic imaging microscopy of normal human and keratoconus cornea
Second-harmonic imaging microscopy of normal human and keratoconus cornea
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DOI：
10.1167/iovs.06-1177
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发表时间：
2007-03-01
影响因子：
4.4
通讯作者：
Jester, James V.
中科院分区：
文献类型：
--
作者：
Morishige, Naoyuki;Wahlert, Andrew J.;Jester, James V.
目的.本研究的目的是评估二次谐波成像识别正常人和圆锥角膜之间角膜基质胶原组织差异的能力。对6例眼库捐献者的正常角膜和13例圆锥角膜患者的穿透性角膜移植术后的角膜进行了检查。使用具有800 nm输出的飞秒钛-蓝宝石激光器产生在400 nm处从中央和旁中央角膜组织块收集的二次谐波信号。收集和重建三维（3-D）数据集，以评估基质胶原板层的位置和方向。二次谐波信号的成像结合正常角膜的3-D重建确定了高度的板层交织，特别是在前角膜。值得注意的是，检测到插入Bowman层并横向于角膜表面定向的椎板，向后穿透约120 μ m。在圆锥角膜中，二次谐波信号成像显示13例中有12例的角膜板层交织较少，且插入Bowman层的角膜板层明显减少或丢失，特别是在与锥体发育相关的区域，Bowman层或瘢痕没有断裂。与正常成人角膜相比，圆锥角膜的角膜前胶原层的结构在二次谐波成像中有明显的异常。这些结构异常与圆锥角膜的胶原组织和生物力学强度的已知变化一致。
PURPOSE. The purpose of this study was to evaluate the ability of second-harmonic imaging to identify differences in corneal stromal Collagen organization between normal human and keratoconus corneas.METHODS. Six normal corneas from eye bank donors and 13 corneas of patients with keratoconus, obtained after penetrating keratoplasty were examined. A femtosecond titanium-sapphire laser with 800-nm output was used to generate second-harmonic signals collected at 400 nm from central and paracentral corneal tissue blocks. Three-dimensional (3-D) data sets were collected and reconstructed to evaluate the location and orientation of stromal collagen lamellae.RESULTS. Imaging of second-harmonic signals combined with 3-D reconstruction of the normal cornea identified a high degree of lamellar interweaving, particularly in the anterior cornea. Of note was the detection of lamellae that inserted into Bowman's layer and were oriented transverse to the corneal surface, penetrating posteriorly approximately 120 mu m. In keratoconus corneas, imaging second-harmonic signals identified less lamellar interweaving and a marked reduction or loss of lamellae inserting into Bowman's layer in 12 of 13 cases, particularly in regions associated with cone development without breaks in Bowman's layer or scarring.CONCLUSIONS. Compared with normal adult corneas, marked abnormalities were detected in the organization of the anterior corneal Collagen lamellae of keratoconus corneas by second harmonic imaging. These structural abnormalities are consistent with the known changes in Collagen organization and biomechanical strength of keratoconus.