Integral Geometry Analysis of Fluorescence Micrographs for Quantitative Relative Comparison of Protein Adsorption onto Polymer Surfaces

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• 作者：Joanna Zemł ; a ; Mał ; gorzata Lekka ; Joanna Wiltowska-Zuber ; Andrzej Budkowski ; Jakub Rysz ; Joanna Raczkowska
• 刊名：Langmuir
• 出版年：2008
• 出版时间：September 16, 2008
• 年：2008
• 卷：24
• 期：18
• 页码：10253-10258
• 全文大小：257K
• 年卷期：v.24,no.18(September 16, 2008)
• ISSN：1520-5827

文摘

Most methods developed to study protein binding to distinct surfaces can only determine the average amount of adsorbed protein or merely provide (qualitative) information on its spatial distribution. Both these features can be characterized rigorously by integral geometry analysis of fluorescence micrographs. This approach is introduced here to compare the relative protein adsorption onto various polymer surfaces: polystyrene (PS), poly(methyl methacrylate) (PMMA), poly(n-butyl methacrylate) (PnBMA), poly(tert-butyl methacrylate) (PtBMA), and PS(PETA) and cross-linked poly(ethylene oxide) (PEO*(PETA)), admixed with pentaerythritol triacrylate (PETA). The polymeric surfaces were incubated for 15 min in phosphate-buffered saline (pH 7.4) containing 125 μg/mL fluorescently labeled lectins, either lentil lectin (LcH) or concanavalin A (ConA). Fluorescence images were recorded at identical conditions (physiological buffer, same exposure time, magnification, gain). For each image, taken a few times for each polymer, the distribution and average value of the normalized intensity were determined. The results show that the binding of LcH to PS(PETA), PtBMA, PS, PnBMA, PMMA, and PEO*(PETA) can be expressed by the ratio of the following values (mean ± 95% confidence interval): 0.356 ± 0.022, 0.298 ± 0.030, 0.241 ± 0.014, 0.083 ± 0.008, 0.039 ± 0.008, and 0.010 ± 0.006, respectively. In turn, the relative adsorption of ConA is described by the values 0.252 ± 0.016, 0.217 ± 0.014, 0.222 ± 0.016, 0.046 ± 0.006, 0.116 ± 0.008, and 0.006 ± 0.002, respectively. Low dispersions of fluorescence intensity around average values indicate homogeneous distribution of adsorbed proteins. The introduced approach enables a fast and easy way not only to quantify the relative amount of bound proteins but also to characterize quantitatively the organization of their surface distribution, as demonstrated for patchlike protein adsorption onto the polymer blend surface.