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{"title":"Microcontact Peeling: A Cell Micropatterning Technique for Circumventing Direct Adsorption of Proteins to Hydrophobic PDMS","authors":"Sho Yokoyama, Tsubasa S. Matsui, Shinji Deguchi","doi":"10.1002/cpcb.22","DOIUrl":null,"url":null,"abstract":"<p>Microcontact printing (μCPr) is one of the most popular techniques used for cell micropatterning. In conventional μCPr, a polydimethylsiloxane (PDMS) stamp with microfeatures is used to adsorb extracellular matrix (ECM) proteins onto the featured surface and transfer them onto particular areas of a cell culture substrate. However, some types of functional proteins other than ECM have been reported to denature upon direct adsorption to hydrophobic PDMS. Here we describe a detailed protocol of an alternative technique––microcontact peeling (μCPe)––that allows for cell micropatterning while circumventing the step of adsorbing proteins to bare PDMS. This technique employs microfeatured materials with a relatively high surface energy such as copper, instead of using a microfeatured PDMS stamp, to peel off a cell-adhesive layer present on the surface of substrates. Consequently, cell-nonadhesive substrates are exposed at the specific surface that undergoes the physical contact with the microfeatured material. Thus, although μCPe and μCPr are apparently similar, the former does not comprise a process of transferring biomolecules through hydrophobic PDMS. © 2017 by John Wiley & Sons, Inc.</p>","PeriodicalId":40051,"journal":{"name":"Current Protocols in Cell Biology","volume":"75 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2017-06-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpcb.22","citationCount":"2","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Cell Biology","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpcb.22","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
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Abstract
Microcontact printing (μCPr) is one of the most popular techniques used for cell micropatterning. In conventional μCPr, a polydimethylsiloxane (PDMS) stamp with microfeatures is used to adsorb extracellular matrix (ECM) proteins onto the featured surface and transfer them onto particular areas of a cell culture substrate. However, some types of functional proteins other than ECM have been reported to denature upon direct adsorption to hydrophobic PDMS. Here we describe a detailed protocol of an alternative technique––microcontact peeling (μCPe)––that allows for cell micropatterning while circumventing the step of adsorbing proteins to bare PDMS. This technique employs microfeatured materials with a relatively high surface energy such as copper, instead of using a microfeatured PDMS stamp, to peel off a cell-adhesive layer present on the surface of substrates. Consequently, cell-nonadhesive substrates are exposed at the specific surface that undergoes the physical contact with the microfeatured material. Thus, although μCPe and μCPr are apparently similar, the former does not comprise a process of transferring biomolecules through hydrophobic PDMS. © 2017 by John Wiley & Sons, Inc.
微接触剥落:一种避免蛋白质在疏水性PDMS上直接吸附的细胞微图技术
微接触印刷(μCPr)是细胞微图像化最常用的技术之一。在传统的μCPr中,使用具有微特征的聚二甲基硅氧烷(PDMS)图章将细胞外基质(ECM)蛋白质吸附到特征表面并将其转移到细胞培养底物的特定区域。然而,据报道,除ECM外,某些类型的功能蛋白在直接吸附到疏水性PDMS后会变性。在这里,我们描述了一种替代技术的详细方案-微接触剥离(μCPe) -允许细胞微图案,同时绕过将蛋白质吸附到裸PDMS的步骤。该技术采用具有相对高表面能的微特征材料,如铜,而不是使用微特征PDMS印记,剥离存在于基板表面的细胞粘附层。因此,细胞非粘附基板暴露在与微特征材料发生物理接触的特定表面。因此,尽管μCPe和μCPr表面上相似,但前者不包含通过疏水PDMS转移生物分子的过程。©2017 by John Wiley &儿子,Inc。
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