The functional activity and differentiation potential of cells is determined by their interaction with surrounding cells. Approaches that allow the unbiased characterization of cell states while at the same time providing spatial information are of major value to assess this environmental influence. However, most current techniques are hampered by a trade-off between spatial resolution and cell profiling depth. Here, we developed a photoswitch-based technology that allows the isolation and in-depth analysis of live cells from regions of interest in complex ex vivo systems, including human tissues. The use of a highly sensitive 4-nitrophenyl(benzofuran)-cage coupled to nanobodies allowed photoswitching of cells in areas of interest with low-intensity violet light and without detectable phototoxicity. Single cell RNA sequencing of spatially defined CD8+ T cells was used to exemplify the feasibility of identifying location-dependent cell states at the single cell level. Finally, we demonstrate the efficient labeling and photoswitching of cells in live primary human tumor tissue. The technology described here provides a valuable tool for the analysis of spatially defined cells in diverse biological systems, including clinical samples.

中文翻译：

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使用新型光开关标签对感兴趣区域（SCARI）进行单细胞分析

细胞的功能活性和分化潜力取决于它们与周围细胞的相互作用。允许对细胞状态进行无偏性表征，同时提供空间信息的方法，对于评估这种环境影响具有重要价值。但是，大多数当前技术都受到空间分辨率和像元分析深度之间折衷的阻碍。在这里，我们开发了一种基于光开关的技术，该技术可以对活体细胞从复杂的离体系统（包括人体组织）中的感兴趣区域进行分离和深入分析。高灵敏度的4-硝基苯基（苯并呋喃）笼与纳米抗体耦合的使用允许在感兴趣的区域中用低强度的紫光对细胞进行光开关，并且没有可检测到的光毒性。使用空间定义的CD8 + T细胞的单细胞RNA测序来举例说明在单细胞水平鉴定位置依赖性细胞状态的可行性。最后，我们证明了活的原发性人类肿瘤组织中细胞的有效标记和光交换。此处描述的技术为分析包括临床样本在内的各种生物系统中空间定义的细胞提供了有价值的工具。