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酿酒酵母黏附的原子力显微镜研究

Atomic Force Microscopy Study of the Adhesion of Saccharomyces cerevisiae.

作者信息

Bowen W. Richard, Lovitt Robert W., Wright Chris J.

机构信息

Centre for Complex Fluids Processing, Department of Chemical and Biological Process Engineering, University of Wales Swansea, Swansea, SA2 8PP, United Kingdom

出版信息

J Colloid Interface Sci. 2001 May 1;237(1):54-61. doi: 10.1006/jcis.2001.7437.

Abstract

An atomic force microscope (AFM) has been used to quantify directly the adhesion of metabolically active Saccharomyces cerevisiae cells at a hydrophilic mica surface, a mica surface with a hydrophobic coating, and a protein-coated mica surface in an aqueous environment. The measurements used "cell probes" constructed by immobilizing a single cell at the apex of a tipless AFM cantilever. Adhesion was quantified from force-distance data for the retraction of the cell from the surface. The data indicated stretching and sequential bond-breaking as the cell probe was retracted from all of the surfaces. Detailed studies were made for physiologically active cells, which were shown to have different adhesion properties to glutaraldehyde-treated cells. Greatest cell adhesion was measured at the hydrophobic surface. Prior adsorption of a bovine serum albumin protein layer at the hydrophilic surface did not significantly affect cell adhesion. Changes in yeast surface hydrophobicity and zeta-potential with yeast cell age were correlated with differences in adhesion. Cells from the stationary phase adhered most strongly to a mica surface. Time of surface contact was demonstrated to be important. Both the force needed to detach a cell from a hydrophilic mica surface and the length of the adhesive interaction increased after 5 min contact. The AFM cell probe technique gives unique insights into primary colonization events in biofilm formation. It will continue to aid both fundamental studies and the assessment of new procedures that are designed to lower cell adhesion at surfaces relevant to biotechnology, medicine, and dentistry Copyright 2001 Academic Press.

摘要

原子力显微镜(AFM)已被用于在水性环境中直接量化代谢活跃的酿酒酵母细胞在亲水性云母表面、具有疏水涂层的云母表面和蛋白质包被的云母表面上的粘附力。测量使用通过将单个细胞固定在无尖端AFM悬臂梁的顶端构建的“细胞探针”。通过细胞从表面缩回的力-距离数据对粘附力进行量化。数据表明,当细胞探针从所有表面缩回时,会发生拉伸和连续的键断裂。对生理活性细胞进行了详细研究,结果表明它们与戊二醛处理的细胞具有不同的粘附特性。在疏水表面测得的细胞粘附力最大。在亲水性表面预先吸附牛血清白蛋白蛋白层对细胞粘附没有显著影响。酵母表面疏水性和ζ电位随酵母细胞年龄的变化与粘附差异相关。稳定期的细胞与云母表面的粘附力最强。表面接触时间被证明很重要。在接触5分钟后,将细胞从亲水性云母表面分离所需的力以及粘附相互作用的长度都增加了。AFM细胞探针技术为生物膜形成中的初始定植事件提供了独特的见解。它将继续有助于基础研究以及评估旨在降低细胞在与生物技术、医学和牙科相关的表面上的粘附力的新程序 版权所有2001年学术出版社。

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