Bogusławski Jakub, Tomczewski Sławomir, Dąbrowski Michał, Komar Katarzyna, Milkiewicz Jadwiga, Palczewska Grażyna, Palczewski Krzysztof, Wojtkowski Maciej
International Centre for Translational Eye Research, Skierniewicka 10A, 01-230 Warsaw, Poland; Institute of Physical Chemistry, Polish Academy of Sciences, Kasprzaka 44/52, 01-224 Warsaw, Poland; Laser and Fiber Electronics Group, Faculty of Electronics, Photonics and Microsystems, Wrocław University of Science and Technology, Wybrzeże Wyspiańskiego 27, 50-370 Wrocław, Poland.
International Centre for Translational Eye Research, Skierniewicka 10A, 01-230 Warsaw, Poland; Institute of Physical Chemistry, Polish Academy of Sciences, Kasprzaka 44/52, 01-224 Warsaw, Poland.
STAR Protoc. 2023 Apr 13;4(2):102225. doi: 10.1016/j.xpro.2023.102225.
Noninvasive imaging of endogenous retinal fluorophores, including vitamin A derivatives, is vital to developing new treatments for retinal diseases. Here, we present a protocol for obtaining in vivo two-photon excited fluorescence images of the fundus in the human eye. We describe steps for laser characterization, system alignment, positioning human subjects, and data registration. We detail data processing and demonstrate analysis with example datasets. This technique allays safety concerns by allowing for the acquisition of informative images at low laser exposure. For complete details on the use and execution of this protocol, please refer to Bogusławski et al. (2022)..
对内源性视网膜荧光团（包括维生素A衍生物）进行无创成像，对于开发视网膜疾病的新疗法至关重要。在此，我们展示了一种获取人眼眼底体内双光子激发荧光图像的方案。我们描述了激光表征、系统校准、人类受试者定位和数据配准的步骤。我们详细介绍了数据处理，并通过示例数据集进行了分析演示。该技术通过在低激光照射下获取信息丰富的图像，减轻了安全担忧。有关此方案的使用和执行的完整详细信息，请参考博古斯拉夫斯基等人（2022年）的研究。
