Okamoto Kazuki, Ebina Teppei, Fujii Naoki, Konishi Kuniaki, Sato Yu, Kashima Tetsuhiko, Nakano Risako, Hioki Hiroyuki, Takeuchi Haruki, Yumoto Junji, Matsuzaki Masanori, Ikegaya Yuji
Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo, Japan.
Department of Cell Biology and Neuroscience, Juntendo University Graduate School of Medicine, Tokyo, Japan.
Sci Adv. 2021 Jan 6;7(2). doi: 10.1126/sciadv.abd2529. Print 2021 Jan.
Optical investigation and manipulation constitute the core of biological experiments. Here, we introduce a new borosilicate glass material that contains the rare-earth ion terbium(III) (Tb), which emits green fluorescence upon blue light excitation, similar to green fluorescent protein (GFP), and thus is widely compatible with conventional biological research environments. Micropipettes made of Tb-doped glass allowed us to target GFP-labeled cells for single-cell electroporation, single-cell transcriptome analysis (Patch-seq), and patch-clamp recording under real-time fluorescence microscopic control. The glass also exhibited potent third harmonic generation upon infrared laser excitation and was usable for online optical targeting of fluorescently labeled neurons in the in vivo neocortex. Thus, Tb-doped glass simplifies many procedures in biological experiments.
光学研究与操控是生物学实验的核心。在此，我们引入一种新型硼硅酸盐玻璃材料，其含有稀土离子铽（III）（Tb），在蓝光激发下会发出绿色荧光，类似于绿色荧光蛋白（GFP），因此与传统生物学研究环境具有广泛的兼容性。由掺Tb玻璃制成的微量移液器使我们能够在实时荧光显微镜控制下，针对GFP标记的细胞进行单细胞电穿孔、单细胞转录组分析（Patch-seq）和膜片钳记录。这种玻璃在红外激光激发下还表现出强大的三次谐波产生能力，可用于体内新皮层中荧光标记神经元的在线光学靶向。因此，掺Tb玻璃简化了生物学实验中的许多程序。
