Abstract
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.
Highlights
Dynamic adhesion and detachment of subcellular regions occur during cell surfaces.[2b,3] by controlling spatial distribution of cell adhesive and migration, a technique allowing precise control of subcellular detachrepellent moieties using microfabricament of cells will be useful for cell migration study
interference reflection microscopy (IRM) images confirmed that PDMP thin film on the spatially modulated light (SML)-illuminated region was dissolved, but MDCK cells on the PDMP-dissolved region remain suspended on the surfaces as shown in differential interference contrast (DIC) images, presumably by tight cell–cell junctions
Force sensing and transmission occurs through focal adhesion complex (FAC),[24] detachment of integrin associated with certain FACs could lead to the movement of FACs toward cell bodies to release tensions applied to the FACs via filamentous actin (F-actin)
Summary
To create surfaces that initially promote cell adhesion, but can trigger partial detachment of cells by light illumination, the surfaces of PDMP, a cell friendly photoresist polymer previously developed for dynamic cell micropatterning (Figure 1A),[11–13,16] was modified. To activate PDMP surfaces, they were treated with air plasma for 1 min prior to fibronectin coating. Www.advancedscience.com substantial reduction of the C–O peak (≈286 eV) that are mostly generated from the (CH2CH2O)n occurred for plasma-treated PDMP surfaces, indicating PEG side chains critical for protein resistance were damaged by plasma treatment. Plasma treatment was sufficient to induce substantial cell adhesion in terms of cell density, presumably by promoting adhesion mole cule binding in FBS on the plasma-treated PDMP surfaces.[17]. Cells on fibronectin-coated PDMP surfaces exhibited more spread morphologies (Figure 1C) with significantly larger areas (Figure 1E) compared with cells on uncoated surfaces (or only plasma-treated surfaces), meaning fibronectin coating on PDMP surfaces further enhanced cell adhesion
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