中国激光, 2022, 49 (24): 2407104, 网络出版: 2022-11-09
基于多光子聚合微笼阵列的单细胞捕获方法 
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Single Cell Capture Method Based on Multiphoton Polymerization Microcage Arrays
图 & 表
图 1. 单脉冲多光子聚合实验原理以及加工的微柱阵列。(a)单脉冲多光子聚合原理示意图，右边虚线框内为倒置样品加工过程的细节放大图，红色椭圆形区域代表聚焦光斑的能量分布，黄色区域表示光刻胶中由聚焦光斑引起的多光子聚合区域；(b)(c)单脉冲多光子聚合加工得到的高度为5 μm和40 μm的4×4微柱阵列；(d)入射激光功率为0.4～0.8 W时，微柱 直径随高度的变化曲线
Fig. 1. Schematic illustrations of single-femtosecond-laser-pulse-based multiphoton polymerization principle and processed micropillars arrays. (a) Schematic illustrations of single-femtosecond-laser-pulse-based multiphoton polymerization principle, where dotted box on the right shows an enlarged view of the processing of inverted sample, red ellipse area represents energy distribution of laser focal spot, and yellow area represents multiphoton polymerized region caused by the corresponding focal spot in the photoresist; (b)(c) 4×4 micropillar arrays with height of 5 μm and 40 μm processed by single-femtosecond-laser-pulse-based multiphoton polymerization; (d) relationship between micropillar diameter and height when laser power ranges from 0.4 W to 0.8 W
图 2. 基于毛细力自组装的图案化微结构阵列，其中加工微柱的激光功率均为0.4 W。(a)临界状态时微柱不稳定自组装结果；(b)由4个微柱自组装形成的微笼结构阵列，右上角为单个微笼的侧视放大图；(c)由8个微柱自组装形成的“四叶草”微结构阵列，右上角为侧视放大图；(d)以(b)中自组装结构为子单元加工的“SIA”图案化结构
Fig. 2. Patterned microstructural arrays based on capillary force self-assembly. (a) Unstable self-assembly of micropillars in critical state; (b) microcage arrays assembled by four micropillars, the enlarged view of the microcage structure is shown at upper right corner; (c)“four-leaf clover”microstructure array assembled by eight micropillars, the enlarged view is shown the upper right corner; (d)“SIA”patterned structure consisted by the self-assembled structure in (b) as a subunit
图 3. 基于毛细力自组装的微球原位捕获。(a)捕获微球的过程示意图；(b)在液体环境中，微球落入微结构的光学图像；(c)在空气环境中，微球被捕获后的光学图像；(d)捕获微球的SEM图，红色虚线圆环中是被捕获的微球
Fig. 3. In situ capture of microspheres based on capillary force self-assembly. (a) Diagram of the process of capturing microspheres; (b) optical image of microsphere falling into microstructures in liquid; (c) optical image of captured microspheres in the air; (d) SEM images of microspheres captured, where the captured microsphere is in the red dotted ring
图 4. 单细胞阵列原位捕获实验步骤示意图。(a)微结构阵列的扫描加工；(b)微结构倒置显影；(c)细胞接种到微结构样本上；(d)细胞贴壁到样本表面；(e)对样本进行细胞消化处理后得到的单细胞阵列；(f)单细胞阵列活死染色；(g)毛细力自组装实现单细胞阵列的原位捕获
Fig. 4. Schematics of single cell array in situ capture experiment procedure. (a) Scanning fabrication of micropillar arrays; (b) inverted development of micropillar structures; (c) cell seeding on microstructural samples; (d) cells adhering to the sample surface; (e) single cell array obtained after cell digestion process; (f) single cell array processed by living dead staining; (g) in situ capture of single cell array by capillary-force-based self-assembly of micropillars
图 5. 微笼阵列的单细胞捕获。(a)MCF-7细胞；(b)(c)除气操作处理前后微结构阵列的光学图像；(d)未接种细胞前，培养液中六微柱微笼阵列的光学图像；(e)接种细胞后，样本表面结构和细胞相对分布的光学图像；(f)消化处理后得到的细胞荧光图像
Fig. 5. Single cell capture in microcage arrays. (a) MCF-7 cells; (b)(c) optical images of microstructure array before and after degassing operation processing; (d) optical image of six micropillars microcage array in culture medium before cell seeding; (e) optical image of sample surface structure and relative distribution of cells after cell seeding; (f) fluorescence images of cells obtained after digestion
图 6. 细胞原位捕获图。(a)(b)(c)四微柱结构原位捕获细胞的荧光图像、光学图像及SEM图；(d)(e)(f)六微柱结构原位捕获细胞的荧光图像、光学图像及SEM图；(g)(h)(i)八微柱结构原位捕获细胞的荧光图像、光学图像及SEM图
Fig. 6. Cells in situ capture diagram. (a)(b)(c) Fluorescence image, optical image and SEM image of cells in situ capture by four micropillars structures; (d)(e)(f) fluorescence image, optical image and SEM image of cells in situ capture by six micropillars structures; (g)(h)(i) fluorescence image, optical image and SEM image of cells in situ capture by eight micropillars structures
杨婷, 孙丽娜, 代国朋, 吕孝峰, 王晓朵. 基于多光子聚合微笼阵列的单细胞捕获方法[J]. 中国激光, 2022, 49(24): 2407104. Ting Yang, Lina Sun, Guopeng Dai, Xiaofeng Lü, Xiaoduo Wang. Single Cell Capture Method Based on Multiphoton Polymerization Microcage Arrays[J]. Chinese Journal of Lasers, 2022, 49(24): 2407104.
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