Flaum Eye Institute, University of Rochester, Rochester, NY, USA.
J Neurophysiol. 2013 May;109(9):2415-21. doi: 10.1152/jn.01043.2012. Epub 2013 Feb 13.
This study reports development of a novel method for high-resolution in vivo imaging of the function of individual mouse retinal ganglion cells (RGCs) that overcomes many limitations of available methods for recording RGC physiology. The technique combines insertion of a genetically encoded calcium indicator into RGCs with imaging of calcium responses over many days with FACILE (functional adaptive optics cellular imaging in the living eye). FACILE extends the most common method for RGC physiology, in vitro physiology, by allowing repeated imaging of the function of each cell over many sessions and by avoiding damage to the retina during removal from the eye. This makes it possible to track changes in the response of individual cells during morphological development or degeneration. FACILE also overcomes limitations of existing in vivo imaging methods, providing fine spatial and temporal detail, structure-function comparison, and simultaneous analysis of multiple cells.
本研究报告了一种新的方法，用于高分辨率的活体成像单个小鼠视网膜神经节细胞（RGCs）的功能，该方法克服了现有 RGC 生理学记录方法的许多限制。该技术将遗传编码的钙指示剂插入到 RGCs 中，然后使用 FACILE（活体眼的功能自适应光学细胞成像）对钙反应进行多天的成像。FACILE 通过允许在多个会话中重复对每个细胞的功能进行成像，并且在从眼睛中取出时避免对视网膜造成损伤，从而扩展了最常见的 RGC 生理学方法，即体外生理学。这使得跟踪单个细胞在形态发育或变性过程中的反应变化成为可能。FACILE 还克服了现有活体成像方法的限制，提供了精细的时空细节、结构-功能比较以及对多个细胞的同时分析。
