Opt Lett. 2024 Jun 15;49(12):3312-3315. doi: 10.1364/OL.525945.
Systems that can image in three dimensions at cellular resolution and across different locations within an organism may enable insights into complex biological processes, such as immune responses, for which a single location measurement may be insufficient. In this Letter, we describe an in vivo two-site imaging probe (TIP) that can simultaneously image two anatomic sites with a maximum separation of a few centimeters. The TIP consists of two identical bendable graded index (GRIN) lenses and is demonstrated by a two-photon two-color fluorescence imaging system. Each GRIN lens has a field of view of 162 ×162 ×170 µm, a nominal numerical aperture of 0.5, a magnification of 0.7, and working distances of 0.2 mm in air for both ends. A blind linear unmixing algorithm is applied to suppress bleedthrough between channels. We use this system to successfully demonstrate two-site two-photon two-color imaging of two biomedically relevant samples, i.e., (1) a mixture of two autofluorescent anti-cancer drugs and (2) a live hybrid tumor consisting of two spectrally distinct fluorescent cell lines.
能够以细胞分辨率在三维空间和生物体的不同位置成像的系统，可以使我们深入了解复杂的生物过程，例如免疫反应，对于这些过程，单次位置测量可能是不够的。在这封信件中，我们描述了一种可同时对两个解剖部位进行成像的体内双位点成像探针（TIP），其最大分离距离可达几厘米。TIP 由两个相同的可弯曲梯度折射率（GRIN）透镜组成，并通过双光子双色荧光成像系统进行了演示。每个 GRIN 透镜的视场为 162×162×170μm，标称数值孔径为 0.5，放大倍率为 0.7，在空气中的工作距离为两端的 0.2mm。我们应用了一种盲线性去混合算法来抑制通道之间的串扰。我们使用该系统成功地演示了两个生物医学相关样本的双位点双光子双色成像，即（1）两种自发荧光抗癌药物的混合物，以及（2）由两个光谱明显不同的荧光细胞系组成的活杂交肿瘤。
