Quantitative three-dimensional confocal imaging of the cornea in situ and in vivo: system design and calibration.
Quantitative three-dimensional confocal imaging of the cornea in situ and in vivo: system design and calibration.
复制标题
DOI：
10.1002/sca.1996.4950180107
复制
发表时间：
2006-12
期刊：
影响因子：
--
通讯作者：
W. Petroll;J. V. Jester;H. D. Cavanagh
中科院分区：
文献类型：
--
作者：
W. Petroll;J. V. Jester;H. D. Cavanagh
提出了一种新的深度编码系统（DES），它可以计算，显示和记录的z轴位置连续在体内成像过程中使用串联扫描共焦显微镜（TSCM）。为了验证DES在体外和体内计算角膜焦平面位置的准确性，我们将角膜厚度的TSCM测量值与使用超声测厚仪（UP，标准临床仪器）在摘除的兔、猫和人眼（n = 15）以及两名人类患者（n = 7）中进行的测量值进行了比较。在摘除眼的UP和TSCM测量值之间发现非常接近的一致性;平均百分比差异为0.50 +/- 2.58%（平均值+/- SD，不显著）。UP和TSCM测量值之间存在显著相关性（R = 0.995，n = 15，p < 0.01）。这些结果验证了由TSCM制造商提供的用于计算焦深的理论方程对于角膜成像是准确的。同样，发现体内UP和TSCM测量值之间非常一致;平均百分比差异为1.67 +/- 1.38%（不显著），证实通过适当压平物镜可以最大限度地减少z轴漂移。这些结果证实了DES用于离体和体内角膜成像的准确性。这个系统应该是非常实用的应用程序中的细胞结构的三维位置的定量是必要的。
A new depth encoding system (DES) is presented, which makes it possible to calculate, display, and record the z-axis position continuously during in vivo imaging using tandem scanning confocal microscopy (TSCM). In order to verify the accuracy of the DES for calculating the position of the focal plane in the cornea both in vitro and in vivo, we compared TSCM measurements of corneal thickness to measurements made using an ultrasonic pachymeter (UP, a standard clinical instrument) in both enucleated rabbit, cat, and human eyes (n = 15), and in both human patients (n = 7). Very close agreement was found between the UP and TSCM measurements in enucleated eyes; the mean percent difference was 0.50 +/- 2.58% (mean +/- SD, not significant). A significant correlation (R = 0.995, n = 15, p < 0.01) was found between UP and TSCM measurements. These results verify that the theoretical equation for calculating focal depth provided by the TSCM manufacturer is accurate for corneal imaging. Similarly, close agreement was found between the in vivo UP and TSCM measurements; the mean percent differences was 1.67 +/- 1.38% (not significant), confirming that z-axis drift can be minimized with proper applanation of the objective. These results confirm the accuracy of the DES for imaging of the cornea both ex vivo and in vivo. This system should be of great utility for applications where quantitation of the three-dimensional location of cellular structures is needed.