The measurement of Bacillus mycoides spore adhesion using atomic force microscopy, simple counting methods, and a spinning disk technique
The measurement of Bacillus mycoides spore adhesion using atomic force microscopy, simple counting methods, and a spinning disk technique
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DOI：
10.1002/bit.10321
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发表时间：
2002-07-20
影响因子：
3.8
通讯作者：
Wright, CJ
中科院分区：
文献类型：
--
作者：
Bowen, WR;Fenton, AS;Wright, CJ
用原子力显微镜研究了蕈状芽孢杆菌孢子在亲水性玻璃表面和疏水性玻璃表面上的粘附。附着在疏水涂层云母表面的孢子的AFM图像允许在没有干燥的水性环境中测量孢子尺寸。观察到孢子外孢壁是柔性的，并且通过增加孢子与表面接触的面积来促进孢子的粘附。使用光学显微镜计数程序的结果与AFM在疏水涂层玻璃表面上观察到的孢子密度相匹配。然而，用AFM在亲水性玻璃表面上没有观察到孢子，这是孢子在该表面上较弱粘附的结果。原子力显微镜也被用来直接定量的相互作用的B。在水性环境中，在两个表面上存在丝状孢子。测量使用的是通过将单孢子固定在无尖端原子力显微镜悬臂梁的顶端而构建的“孢子探针”。数据显示，当孢子从亲水性玻璃表面缩回时，发生拉伸和连续的键断裂。最大的孢子粘附在疏水涂层的玻璃表面测量。当孢子接近疏水涂覆的表面时，测量孢子上的吸引力。在亲水性玻璃表面，在孢子接近期间仅测量排斥力。的AFM力的测量是在定性协议的流体动力学剪切粘附试验，使用旋转盘技术的结果。从定量上讲，AFM测量的粘附力比使用旋转盘数据进行的估计大4 × 10（3）倍。这是在不同粘附试验中对孢子施加不同类型力的结果。原子力显微镜提供了一些独特的见解孢子与表面的相互作用。没有其他仪器可以对单个微生物细胞进行这样的直接测量。(C)2002 Wiley Periodicals，Inc.
An atomic force microscope has been used to study the adhesion of Bacillus mycoides spores to a hydrophilic glass surface and a hydrophobic-coated glass surface. AFM images of spores attached to the hydrophobic-coated mica surface allowed the measurement of spore dimensions in an aqueous environment without desiccation. The spore exosporium was observed to be flexible and to promote the adhesion of the spore by increasing the area of spore contact with the surface. Results from counting procedures using light microscopy matched the density of spores observed on the hydrophobic-coated glass surface with AFM. However, no spores were observed on the hydrophilic glass surface with AFM, a consequence of the weaker adhesion of the spores at this surface. AFM was also used to quantify directly the interactions of B. mycoides spores at the two surfaces in an aqueous environment. The measurements used "spore probes" constructed by immobilizing a single spore at the apex of a tipless AFM cantilever. The data showed that stretching and sequential bond breaking occurred as the spores were retracted from the hydrophilic glass surface. The greatest spore adhesion was measured at the hydrophobic-coated glass surface. An attractive force on the spores was measured as the spores approached the hydrophobic-coated surface. At the hydrophilic glass surface, only repulsive forces were measured during the approach of the spores. The AFM force measurements were in qualitative agreement with the results of a hydrodynamic shear adhesion assay that used a spinning disk technique. Quantitatively, AFM measurements of adhesive force were up to 4 x 10(3) times larger than the estimates made using the spinning disk data. This is a consequence of the different types of forces applied to the spore in the different adhesion assays. AFM has provided some unique insights into the interactions of spores with surfaces. No other instrument can make such direct measurements for single microbiological cells. (C) 2002 Wiley Periodicals, Inc.