The functional activity and differentiation potential of cells are determined by their interactions with surrounding cells. Approaches that allow 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 tradeoff between spatial resolution and cell profiling depth. Here, we develop a photocage-based technology that allows isolation and in-depth analysis of live cells from regions of interest in complex ex vivo systems, including primary human tissues. The use of a highly sensitive 4-nitrophenyl(benzofuran) cage coupled to a set of nanobodies allows high-resolution photo-uncaging of different cell types in areas of interest. Single-cell RNA-sequencing of spatially defined CD8⁺ T cells is used to exemplify the feasibility of identifying location-dependent cell states. The technology described here provides a valuable tool for the analysis of spatially defined cells in diverse biological systems, including clinical samples.

细胞的功能活性和分化潜力是由它们与周围细胞的相互作用决定的。允许对细胞状态进行公正表征，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。然而，大多数当前技术都受到空间分辨率和细胞分析深度之间的权衡的阻碍。在这里，我们开发了一种基于光笼的技术，可以从复杂的离体系统（包括原代人体组织）的感兴趣区域中分离和深入分析活细胞。使用与一组纳米体耦合的高灵敏度 4-硝基苯基（苯并呋喃）笼可以对感兴趣区域中的不同细胞类型进行高分辨率光解笼。空间限定的 CD8 ⁺ T 细胞的单细胞 RNA 测序用于举例说明识别位置依赖性细胞状态的可行性。这里描述的技术为分析不同生物系统（包括临床样本）中空间定义的细胞提供了一个有价值的工具。