Bohn Sebastian, Stahnke Thomas, Sperlich Karsten, Linke Stephan J, Farrokhi Sanaz, Klemm Maren, Allgeier Stephan, Köhler Bernd, Reichert Klaus-Martin, Witt Martin, Stachs Oliver, Guthoff Rudolf F
Universitätsaugenklinik, Universitätsmedizin Rostock, Deutschland.
Department Leben, Licht & Materie, Universität Rostock, Deutschland.
Klin Monbl Augenheilkd. 2020 Dec;237(12):1442-1454. doi: 10.1055/a-1297-4717. Epub 2020 Nov 23.
Confocal in vivo microscopy is an established method in ophthalmology research. As it requires contact coupling and calibration of the instruments is suboptimal, this method has been only rarely used in clinical routine work. As a result of close collaboration between physicists, information scientists and ophthalmologists, confocal laser scanning microscopy (CLSM) of the eye has been developed in recent years and a prototype can now be used in patients. The present study evaluates possible clinical uses of this method.
The essential innovations in CLSM are (1) a newly designed coupling element with superficial adaptation to corneal curvature and (2) the use of a dual computerised piezo drive for rapid and precise focusing. In post-processing and after elastic imaging registration of the individual images parallel to the surface, it is also possible to produce sagittal sections resembling a split lamp and with resolution in the micrometer range. The concept was tested on enucleated pig bulbi and tested on normal volunteers and selected patients with diseases of the cornea.
Simultaneous imaging in planes parallel to the surface and in sagittal planes provided additional information that can help us to understand the processes of wound healing in all substructures of the cornea and the role of immune competent cells. Possible clinical uses were demonstrated in a volunteer with healthy eyes and several groups of patients (keratoconus after CXL, recurrent keratitis, status after PRK). These show that this new approach can be used in morphological studies at cellular level in any desired and appropriate test plane.
It could be shown that this new concept of CLSM can be used clinically. It can provide valuable and novel information to both preclinical researchers and to ophthalmologists interested in corneal disease, e.g. density of Langerhans cells and epithelial stratification in ocular surface diseases.
共聚焦体内显微镜检查是眼科研究中的一种成熟方法。由于它需要接触耦合且仪器校准不够理想，这种方法在临床常规工作中很少使用。由于物理学家、信息科学家和眼科医生的密切合作，近年来已开发出眼部共聚焦激光扫描显微镜（CLSM），现在有一种原型可用于患者。本研究评估了该方法可能的临床用途。
CLSM的主要创新之处在于：（1）新设计的耦合元件，可根据角膜曲率进行表面适配；（2）使用双计算机控制的压电驱动器进行快速精确聚焦。在对平行于表面的单个图像进行后处理和弹性成像配准后，还可以生成类似裂隙灯的矢状切面，分辨率可达微米级。该概念在摘除的猪眼球上进行了测试，并在正常志愿者和选定的角膜疾病患者身上进行了测试。
在平行于表面的平面和矢状平面上同时成像提供了额外信息，有助于我们了解角膜所有亚结构中的伤口愈合过程以及免疫活性细胞的作用。在一名健康眼睛的志愿者和几组患者（CXL术后圆锥角膜、复发性角膜炎、PRK术后状态）中展示了可能的临床用途。这些表明，这种新方法可用于在任何所需的合适测试平面上进行细胞水平的形态学研究。
可以证明，CLSM的这一新概念可用于临床。它可以为临床前研究人员和对角膜疾病感兴趣的眼科医生提供有价值的新信息，例如眼表疾病中朗格汉斯细胞的密度和上皮分层情况。
