Two-photon imaging of the mouse eye.
Two-photon imaging of the mouse eye.
复制标题
小鼠眼睛的双光子成像。
DOI：
10.1167/iovs.10-7115
复制
发表时间：
2011
影响因子：
4.4
通讯作者：
M. Kahook
中科院分区：
文献类型：
--
作者：
A. Johnson;D. Ammar;M. Kahook
目的
采用无创性显微镜技术对完整摘除的小鼠眼内的常规房水流出途径和邻近结构进行成像。
方法
双光子显微镜（2PM）技术，包括双光子自发荧光（2PAF）和二次谐波产生（SHG），用于获得图像的小梁网（TM）区域内的一个完整的小鼠眼睛。心脏灌注的荧光素结合的葡聚糖被用来标记眼睛内的血管，作为解剖参考。随后将眼睛固定、石蜡包埋、切片并染色，以与2PM图像进行比较。
结果
对多个2PM图像的三维分析显示，虹膜和角膜附近有一个明确的区域，没有SHG信号，与Schlemm管的位置一致。该开放区域与与收集器通道一致的较小的管结构是连续的。这些结构在用血管探针葡聚糖灌注的小鼠中没有标记，支持封闭空间充满房水而不是循环血液的假设。小鼠眼中的TM区域也是可见的，具有代表胶原纤维的清晰SHG信号。
结论
这些结果支持了2PM可用于青光眼小鼠模型中常规房水流出通路的非侵入性成像的假设。研究正在进行中，以验证我们的方法在活体动物。
PURPOSE
To image the conventional aqueous outflow pathway and adjacent structures within the intact enucleated mouse eye using a noninvasive microscopy technique.
METHODS
Two-photon microscopy (2PM) techniques, including two-photon autofluorescence (2PAF) and second harmonic generation (SHG), were used to obtain images of the trabecular meshwork (TM) region within an intact mouse eye. Cardiac perfusion of fluorescein-conjugated dextran was used to label blood vessels within the eye to serve as an anatomic reference. Eyes were subsequently fixed, paraffin embedded, sectioned, and stained for comparison to the 2PM images.
RESULTS
Three-dimensional analyses of multiple 2PM images revealed a well-defined region adjacent to the iris and cornea that is free of SHG signal and consistent with the location of Schlemm's canal. This open region is continuous with smaller tube structures consistent with collector channels. These structures do not label in mice perfused with the vascular probe dextran, supporting the hypothesis that the enclosed spaces are filled with aqueous humor rather than circulating blood. The TM region in the mouse eye was also visible, with a clear SHG signal representing collagen fibers.
CONCLUSIONS
These results support the hypothesis that 2PM may be useful for noninvasively imaging the conventional aqueous outflow pathway in mouse models of glaucoma. Studies are ongoing to validate our methodology in live animals.
登录
查看更多内容
影响因子：
3.4
作者：
Toyran, S;Liu, YM;Edward, DP
通讯作者：
Edward, DP
影响因子：
3.4
作者：
McKinnon, Stuart J.;Schlamp, Cassandra L.;Nickells, Robert W.
通讯作者：
Nickells, Robert W.
影响因子：
2.2
作者：
D. Ammar;Tim C. Lei;E. Gibson;M. Kahook
通讯作者：
D. Ammar;Tim C. Lei;E. Gibson;M. Kahook
影响因子：
4
作者：
Pelz, Philipp M.;Guizar-Sicairos, Manuel;Menzel, Andreas
通讯作者：
Menzel, Andreas
影响因子：
3.4
作者：
MAEPEA, O;BILL, A
通讯作者：
BILL, A