细胞分型是一种强大的技术，可以精确地控制和排列细胞，实现广泛的单细胞分析，广泛应用于治疗、诊断和再生医学。本研究提出了一种新颖高效的技术，能够实现大规模并行高通量细胞分型和准确输送小至大的生物分子到图案化细胞中。本研究提出的创新细胞分型装置是一个独立的超薄3D SU-8微网膜，厚度为10μm，具有40μm至80μm的微孔阵列，孔间距为50μm至150μm。利用该装置，在Petri培养皿上培养细胞，可实现高效的细胞图案，可变为从单个细胞到细胞簇。利用这种技术，我们对小鼠成纤维细胞L929（80.5%）、人宫颈癌SiHa（81%）和人神经母细胞瘤IMR32（89.6%）实现了极高的可重复分型效率，不良区域的缺陷率小于1%。对于L929细胞，单细胞分型效率最高为75.8%。此外，我们通过采用红外光脉冲激发的一系列钛微环（外径10μm，内径3μm）阵列，展示了大规模并行高通量的大分子转染进图案化活细胞的能力。我们成功地将一系列不同大小的生物分子输送到各类癌细胞图案中，包括碘化丙啶（668.4Da）、右旋糖酐（3kDa）、小干扰RNA（13.3kDa）和β-半乳糖苷酶酶（465kDa）。值得注意的是，我们的平台对于诸如碘化丙啶等小分子的输送效率达到97%，对于酶的输送效率达到84%，相应的细胞存活率分别为100%和90%。此外，基于SU-8的紧凑可重复使用的膜装置有助于高效的细胞分型、转染和细胞存活，将成为诊断和治疗应用的有前景的工具。

Cell patterning is a powerful technique for the precise control and arrangement of cells, enabling detailed single-cell analysis with broad applications in therapeutics, diagnostics, and regenerative medicine. This study presents a novel and efficient technique that enables massively parallel high throughput cell patterning and precise delivery of small to large biomolecules into patterned cells. The innovative cell patterning device proposed in this study is a standalone, ultrathin 3D SU-8 micro-stencil membrane, with a thickness of 10 μm. It features an array of micro-holes ranging from 40 μm to 80 μm, spaced apart by 50 μm to 150 μm. By culturing cells on top of this SU-8 membrane, the technique achieves highly efficient cell patterns varying from single-cell to cell clusters on a Petri dish. Utilizing this technique, we have achieved a remarkable reproducible patterning efficiency for mouse fibroblast L929 (80.5%), human cervical SiHa (81%), and human neuroblastoma IMR32 (89.6%) with less than 1% defects in undesired areas. Single-cell patterning efficiency was observed to be highest at 75.8% for L929 cells. Additionally, we have demonstrated massively parallel high throughput uniform transfection of large biomolecules into live patterned cells by employing an array of titanium micro-rings (10 μm outer diameter, 3 μm inner diameter) activated through infrared light pulses. Successful delivery of a wide range of small to very large biomolecules, including propidium iodide (PI) dye (668.4 Da), dextran (3 kDa), siRNA (13.3 kDa), and β-galactosidase enzyme (465 kDa), was accomplished in cell patterns for various cancer cells. Notably, our platform achieved exceptional delivery efficiencies of 97% for small molecules like PI dye and 84% for the enzyme, with corresponding high cell viability of 100% and 90%, respectively. Furthermore, the compact and reusable SU-8-based membrane device facilitates highly efficient cell patterning, transfection, and cell viability, making it a promising tool for diagnostics and therapeutic applications.