Bar-Noam Adi Schejter, Farah Nairouz, Shoham Shy
Faculty of Biomedical Engineering, Technion - Israel Institute of Technology, Kiryat HaTechnion, Haifa 32000, Israel.
Light Sci Appl. 2016 Jan 1;5(1):e16007. doi: 10.1038/lsa.2016.7. eCollection 2016 Jan.
Non-invasive fluorescence retinal imaging in small animals is an important requirement for an array of translational vision applications. The two-photon imaging of the mouse retina may enable the long-term investigation of the structure and function of healthy and diseased retinal tissue. However, to date, this has only been possible using relatively complex adaptive-optics systems. Here, the optical modeling of the murine eye and of the imaging system is used to achieve correction-free two-photon microscopy through the pupil of a mouse eye to yield high-quality, optically sectioned fundus images. By remotely scanning the focus using an electronically tunable lens, high-resolution three-dimensional fluorescein angiograms and cellular-scale images are acquired, thus introducing a correction-free baseline performance level for two-photon retinal imaging. Moreover, the system enables functional calcium imaging of repeated retinal responses to light stimulation using the genetically encoded indicator, GCaMP6s. These results and the simplicity of the new add-on optics are an important step toward several structural, functional, and multimodal imaging applications that will benefit from the tight optical sectioning and the use of near-infrared light.
小动物的非侵入性荧光视网膜成像对于一系列转化视觉应用来说是一项重要需求。小鼠视网膜的双光子成像或许能够对健康和患病视网膜组织的结构与功能进行长期研究。然而，迄今为止，这仅能通过相对复杂的自适应光学系统来实现。在此，利用小鼠眼睛和成像系统的光学建模，通过小鼠眼睛的瞳孔实现无校正双光子显微镜检查，以产生高质量的光学切片眼底图像。通过使用电子可调透镜对焦点进行远程扫描，获取了高分辨率三维荧光血管造影图像和细胞尺度图像，从而为双光子视网膜成像引入了无校正基线性能水平。此外，该系统能够使用基因编码指示剂GCaMP6s对视网膜对光刺激的重复反应进行功能性钙成像。这些结果以及新型附加光学器件的简易性，是迈向若干结构、功能和多模态成像应用的重要一步，这些应用将受益于紧密的光学切片和近红外光的使用。
