Assembly of live micro-organisms on microstructured PDMS stamps by convective/capillary deposition for AFM bio-experiments
Assembly of live micro-organisms on microstructured PDMS stamps by convective/capillary deposition for AFM bio-experiments
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DOI：
10.1088/0957-4484/22/39/395102
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发表时间：
2011-09-30
期刊：
影响因子：
3.5
通讯作者：
Ressier, L.
中科院分区：
文献类型：
--
作者：
Dague, E.;Jauvert, E.;Ressier, L.
将活微生物固定在固体基质上是原子力显微镜生物实验的重要前提。所采用的方法必须足够牢固地固定细胞，使它们能够承受扫描过程中尖端施加的横向摩擦力，但不会使细胞界面变性。在这项工作中，提出了一种在特定底物区域上组装活细胞的通用方法。它包括使用对流/毛细管沉积在微结构、功能化的聚二甲基硅氧烷(PDMS)邮票图案内组装活细胞。这种通用的方法通过将其应用于生物技术和医学中最重要的两个系统来验证：酿酒酵母和烟曲霉真菌孢子。我们证明，这种方法可以通过作用力光谱对酿酒酵母进行多路AFM纳米机械测量，并在缓冲介质中对萌发的曲霉分生孢子进行高分辨率AFM成像。这两个例子清楚地展示了通过对流/毛细管沉积在功能化的微结构PDMS邮票上组装微生物的巨大潜力，用于在活细胞上进行严格的AFM生物实验。
Immobilization of live micro-organisms on solid substrates is an important prerequisite for atomic force microscopy (AFM) bio-experiments. The method employed must immobilize the cells firmly enough to enable them to withstand the lateral friction forces exerted by the tip during scanning but without denaturing the cell interface. In this work, a generic method for the assembly of living cells on specific areas of substrates is proposed. It consists in assembling the living cells within the patterns of microstructured, functionalized poly-dimethylsiloxane (PDMS) stamps using convective/capillary deposition. This versatile approach is validated by applying it to two systems of foremost importance in biotechnology and medicine: Saccharomyces cerevisiae yeasts and Aspergillus fumigatus fungal spores. We show that this method allows multiplexing AFM nanomechanical measurements by force spectroscopy on S. cerevisiae yeasts and high-resolution AFM imaging of germinated Aspergillus conidia in buffer medium. These two examples clearly demonstrate the immense potential of micro-organism assembly on functionalized, microstructured PDMS stamps by convective/capillary deposition for performing rigorous AFM bio-experiments on living cells.