Experimental Designs to Study the Aggregation and Colonization of Biofilms by Video Microscopy With Statistical Confidence.
Experimental Designs to Study the Aggregation and Colonization of Biofilms by Video Microscopy With Statistical Confidence.
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DOI：
10.3389/fmicb.2021.785182
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发表时间：
2021
影响因子：
5.2
通讯作者：
Parker AE
中科院分区：
文献类型：
--
作者：
Pettygrove BA;Smith HJ;Pallister KB;Voyich JM;Stewart PS;Parker AE
本研究的目标是量化早期定殖生物膜的共聚焦激光扫描显微镜（CLSM）延时图像的可变性，以帮助设计未来的成像实验。为了实现这一点，利用了由16个独立的CLSM显微镜实验组成的大型成像数据集。这些实验旨在研究人中性粒细胞与金黄色葡萄球菌（S。金黄色葡萄球菌）。结果表明，在未处理的对照实验中，生长阶段之间的变异性显著不同（即，滞后或指数）。当研究抗微生物处理（在这种情况下，嗜中性粒细胞攻击）的效果时，无论接种水平或生长阶段如何，变异性作为处理功效的皱眉形函数（即，生物膜表面覆盖率的降低）。这些发现被用来预测早期生物膜的未来成像研究的最佳实验设计，通过考虑不同的（i）独立实验的数量;（ii）每个实验的视场（FOV）数量;和（iii）每小时的帧捕获率。包括能够评估任何用户指定设计的电子表格，该电子表格需要来自用户生成的实验结果的预期平均对数减少和方差分量。本研究中概述的方法可以帮助研究人员设计具有高统计置信度的抗菌治疗CLSM研究。
The goal of this study was to quantify the variability of confocal laser scanning microscopy (CLSM) time-lapse images of early colonizing biofilms to aid in the design of future imaging experiments. To accomplish this a large imaging dataset consisting of 16 independent CLSM microscopy experiments was leveraged. These experiments were designed to study interactions between human neutrophils and single cells or aggregates of Staphylococcus aureus (S. aureus) during the initial stages of biofilm formation. Results suggest that in untreated control experiments, variability differed substantially between growth phases (i.e., lag or exponential). When studying the effect of an antimicrobial treatment (in this case, neutrophil challenge), regardless of the inoculation level or of growth phase, variability changed as a frown-shaped function of treatment efficacy (i.e., the reduction in biofilm surface coverage). These findings were used to predict the best experimental designs for future imaging studies of early biofilms by considering differing (i) numbers of independent experiments; (ii) numbers of fields of view (FOV) per experiment; and (iii) frame capture rates per hour. A spreadsheet capable of assessing any user-specified design is included that requires the expected mean log reduction and variance components from user-generated experimental results. The methodology outlined in this study can assist researchers in designing their CLSM studies of antimicrobial treatments with a high level of statistical confidence.
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影响因子：
3.1
作者：
Ghimire, Niranjan;Pettygrove, Brian A.;Stewart, Philip S.
通讯作者：
Stewart, Philip S.
DOI：
10.1007/978-1-4939-0554-6_13
发表时间：
2014-01-01
期刊：
ENGINEERING AND ANALYZING MULTICELLULAR SYSTEMS: METHODS AND PROTOCOLS
影响因子：
--
作者：
Brileya, Kristen A.;Camilleri, Laura B.;Fields, Matthew W.
通讯作者：
Fields, Matthew W.
影响因子：
3.6
作者：
Allesen-Holm, M;Barken, KB;Tolker-Nielsen, T
通讯作者：
Tolker-Nielsen, T
影响因子：
3.8
作者：
Renslow, Ryan;Lewandowski, Zbigniew;Beyenal, Haluk
通讯作者：
Beyenal, Haluk
影响因子：
3.3
作者：
Lee JY;Kitaoka M
通讯作者：
Kitaoka M