Imaging corneal crosslinking by autofluorescence 2‐photon microscopy, second harmonic generation, and fluorescence lifetime measurements
Imaging corneal crosslinking by autofluorescence 2‐photon microscopy, second harmonic generation, and fluorescence lifetime measurements
复制标题
通过自发荧光双光子显微镜、二次谐波产生和荧光寿命测量对角膜交联进行成像
DOI：
10.1016/j.jcrs.2010.06.068
复制
发表时间：
2010
影响因子：
2.8
通讯作者：
O. Stachs
中科院分区：
文献类型：
--
作者：
P. Steven;M. Hovakimyan;R. Guthoff;G. Hüttmann;O. Stachs
目的：评价激发组织自体荧光的双光子显微镜（TPM）在检测和计算胶原角膜交联等级中的应用，胶原角膜交联是通过裂隙灯和体内共聚焦显微镜不可见的。单位：吕贝克大学眼科系，吕贝克，罗斯托克大学，罗斯托克，德国。设计：实验研究。方法：兔角膜用不同的交联（CXL）方案处理。治疗后两周，通过共聚焦显微镜在体内评价角膜。摘除眼睛，并在710 nm和826 nm激发波长下进行TPM，以检测组织自发荧光、二次谐波产生和荧光寿命测量（FLIM）。眼睛，然后固定和分析histologics.RESULTS：交联后的标准协议产生了强烈的自体荧光信号中的基质，检测TPM。该信号在对照样本中较弱，并且在外周区和CXL区之间观察到明显的过渡区。在FLIM，增加角膜交联测量时，标准协议used.CONCLUSIONS：双光子显微镜，一种非侵入性的方法，能够检测到治疗CXL的效果和测量CXL的等级。除了术后治疗控制外，该技术还可用于双光子触发CXL期间的在线剂量测定。
PURPOSE:To evaluate the use of 2-photon microscopy (TPM), which excites tissue autofluorescence, in detecting and calculating the grade of collagen corneal crosslinks, which are not visible through the slitlamp and in vivo confocal microscopy.SETTING:Departments of Ophthalmology, University of Lübeck, Lübeck, and University of Rostock, Rostock, Germany.DESIGN:Experimental study.METHODS:Corneas of rabbits were treated with different crosslinking (CXL) protocols. Two weeks after treatment, the corneas were evaluated in vivo by confocal microscopy. Eyes were enucleated and TPM was performed at 710 nm and 826 nm excitation wavelengths to detect tissue autofluorescence, second harmonic generation, and fluorescence lifetime measurements (FLIM). Eyes were then fixed and analyzed by histology.RESULTS:Crosslinking following the standard protocol generated a strong autofluorescence signal in the stroma that was detected by TPM. This signal was weakly present in the control specimens, and a sharp transition zone between the peripheral zone and the CXL zone was seen. On FLIM, an increase in corneal crosslinks was measured when the standard protocol was used.CONCLUSIONS:Two-photon microscopy, a noninvasive method, was able to detect the effects of therapeutic CXL and measure the grade of CXL. In addition to postoperative treatment control, the technique has possibilities for use in online dosimetry during 2-photon triggered CXL.
登录
查看更多内容
影响因子：
4.4
作者：
Lili Zhou;S. Sawaguchi;S. Twining;J. Sugar;R. Feder;B. J. Yue
通讯作者：
Lili Zhou;S. Sawaguchi;S. Twining;J. Sugar;R. Feder;B. J. Yue
影响因子：
3.4
作者：
Raub, Christopher B.;Suresh, Vinod;George, Steven C.
通讯作者：
George, Steven C.
影响因子：
4.2
作者：
Patel, SV;Hodge, DO;Bourne, WM
通讯作者：
Bourne, WM
影响因子：
56.9
作者：
DENK, W;STRICKLER, JH;WEBB, WW
通讯作者：
WEBB, WW