Measurement of local diffusion coefficients in biofilms by microinjection and confocal microscopy.
Measurement of local diffusion coefficients in biofilms by microinjection and confocal microscopy.
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DOI：
10.1002/(sici)1097-0290(19970120)53:2
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发表时间：
1997-01
影响因子：
3.8
通讯作者：
D. Beer;P. Stoodley;Z. Lewandowski
中科院分区：
文献类型：
--
作者：
D. Beer;P. Stoodley;Z. Lewandowski
介绍了一种测定生物膜局部扩散系数的新方法。它是基于荧光染料的显微注射和使用共聚焦激光显微镜对随后羽流形成的定量分析。测定了荧光素（mw332）、TRITC-IgG （mw150000）和植红蛋白（mw240000）在异质生物膜细胞簇和间隙空隙中的扩散系数。孔洞中测量的扩散系数与水中的理论值接近。荧光素在细胞簇、空隙和无菌培养基中具有相同的扩散性。TRITC-IgG没有在细胞簇中扩散，可能是由于与细胞簇基质结合所致。生物膜经牛血清白蛋白处理后，结合能力下降，扩散系数可测。与细胞间隙相比，藻红蛋白在细胞簇中的扩散率降低了41%。从藻红蛋白的扩散数据进一步计算出细胞簇基质具有凝胶特性，纤维厚度为0.6 nm，孔径为80 nm。(c) 1997 John Wiley & Sons, Inc。
A new technique for the determination of local diffusion coefficients in biofilms is described. It is based on the microinjection of fluorescent dyes and quantitative analysis of the subsequent plume formation using confocal laser microscopy. The diffusion coefficients of fluorescein (MW 332), TRITC-IgG (MW 150000) and phycoerythrin (MW 240000) were measured in the cell clusters and interstitial voids of a heterogeneous biofilm. The diffusivities measured in the voids were close to the theoretical values in water. Fluorescein had the same diffusivity in cell clusters, voids, and sterile medium. TRITC-IgG did not diffuse in cell clusters, presumably due to binding to the cell cluster matrix. After treatment of the biofilm with bovine serum albumin, binding capacity decreased and the diffusion coefficient could be measured. The diffusivity of phycoerythrin in cell clusters was impeded by 41%, compared to interstitial voids. From the diffusion data of phycoerythrin it was further calculated that the cell cluster matrix had the characteristics of a gel with 0.6 nm thick fibers and pore diameters of 80 nm. (c) 1997 John Wiley & Sons, Inc.