Park Jeehun, Kim Taeyup, Choi Jong Chul, Doh Junsang
School of Interdisciplinary Bioscience and Bioengineering (I-Bio) Pohang University of Science and Technology 77, Cheongam-ro Pohang Gyeongbuk 37673 South Korea.
Department of Mechanical Engineering Pohang University of Science and Technology 77, Cheongam-ro Pohang Gyeongbuk 37673 South Korea.
Adv Sci (Weinh). 2019 May 17;6(14):1900566. doi: 10.1002/advs.201900566. eCollection 2019 Jul 17.
Dynamic adhesion and detachment of subcellular regions occur during cell migration, thus a technique allowing precise control of subcellular detachment of cells will be useful for cell migration study. Previous methods for cell detachment were developed either for harvesting cells or cell sheets attached on surfaces with low resolution patterning capability, or for detaching subcellular regions located on predefined electrodes. In this paper, a method that allows in situ subcellular detachment of cells with ≈1.5 µm critical feature size while observing cells under a fluorescence microscope is introduced using a cell-friendly photoresist and spatially modulated light. Using this method, a single cell, regions in cell sheets, and a single focal adhesion complex within a cell are successfully detached. Furthermore, different subcellular regions of migrating cells are detached and changes in cell polarity and migration direction are quantitatively analyzed. This method will be useful for many applications in cell detachment, in particular when subcellular resolution is required.
亚细胞区域的动态黏附和脱离发生在细胞迁移过程中,因此一种能够精确控制细胞亚细胞脱离的技术将有助于细胞迁移研究。先前的细胞脱离方法,要么是用于收获细胞或附着在具有低分辨率图案化能力的表面上的细胞片,要么是用于脱离位于预定义电极上的亚细胞区域。在本文中,介绍了一种方法,该方法使用对细胞友好的光刻胶和空间调制光,在荧光显微镜下观察细胞的同时,能够以约1.5 µm的关键特征尺寸原位实现细胞的亚细胞脱离。使用该方法,成功地分离了单个细胞、细胞片中的区域以及细胞内的单个黏着斑复合体。此外,对迁移细胞的不同亚细胞区域进行了分离,并对细胞极性和迁移方向的变化进行了定量分析。该方法将在细胞脱离的许多应用中发挥作用,特别是在需要亚细胞分辨率的情况下。