A microtiter peg lid with ziggurat geometry for medium-throughput antibiotic testing and in situ imaging of biofilms.

A microtiter peg lid with ziggurat geometry for medium-throughput antibiotic testing and in situ imaging of biofilms.
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用于中等通量抗生素测试和生物膜原位成像的具有金字塔形几何形状的微量滴定柱盖。

DOI:
10.1016/j.bioflm.2023.100167
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发表时间:
2023-12-15
期刊:
影响因子:
6.8
通讯作者:
--
中科院分区:
其他
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--
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使用多种方法量化和观察细菌生物膜对消毒剂和抗生素的反应,但由于速度快、减少用户错误和原位分析,优选显微镜,特别是共焦激光扫描显微镜 (CLSM)。 CLSM 可以以有限的吞吐量解决 3D 生物膜的生物学和空间异质性。 ASTM E2799-22 中描述的基于微孔板钉盖的测定是一种用于测试生物膜的中等通量方法,但不允许进行原位成像。按照制造商的建议折断钉子可能会损坏样品,并且仅限于易于触及的钉子。在这里,我们报告了针对原位可视化和所有钉子可视化而优化的钉子的修改。我们报告了类似的抗生素挑战恢复,通过遵循 ASTM E2799-22 协议和原位成像的集落形成。我们报告了抗生素对生物膜形态,特别是生物膜流带的新的可量化影响。新设计将选择生物膜表型的 MBEC® 检测设计与原位成像需求结合起来。
Bacteria biofilm responses to disinfectants and antibiotics are quantified and observed using multiple methods, though microscopy, particularly confocal laser scanning microscopy (CLSM) is preferred due to speed, a reduction in user error, and in situ analysis. CLSM can resolve biological and spatial heterogeneity of biofilms in 3D with limited throughput. The microplate peg-lid-based assay, described in ASTM E2799-22, is a medium-throughput method for testing biofilms but does not permit in situ imaging. Breaking off the peg, as recommended by the manufacturer, risks sample damage, and is limited to easily accessible pegs. Here we report modifications to the peg optimized for in situ visualization and visualization of all pegs. We report similar antibiotic challenge recovery via colony formation following the ASTM E2799-22 protocol and in situ imaging. We report novel quantifiable effects of antibiotics on biofilm morphologies, specifically biofilm streamers. The new design bridges the MBEC® assays design that selects for biofilm phenotypes with in situ imaging needs.
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