用于中等通量抗生素检测和生物膜原位成像的微型滴度钉盖

IF 5.9 Q1 MICROBIOLOGY Biofilm Pub Date : 2023-11-17 DOI:10.1016/j.bioflm.2023.100167
Sarah K. Childs , A-Andrew D. Jones III
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引用次数: 0

摘要

细菌生物膜对消毒剂和抗生素的反应是用多种方法量化和观察的,尽管显微镜,特别是共聚焦激光扫描显微镜(CLSM)是首选,因为速度快,减少了用户的错误,并且在原位分析。CLSM可以在有限的通量下三维解析生物膜的生物学和空间异质性。在ASTM E2799-22中描述的基于微孔板peg-盖的检测是一种中等通量的生物膜检测方法,但不允许原位成像。如制造商所建议的那样,折断木栓有损坏样品的风险,并且仅限于容易接近的木栓。在这里,我们报告了对peg的修改,优化了所有peg的原位可视化和可视化。我们根据ASTM E2799-22方案和原位成像,通过菌落形成报告类似的抗生素挑战恢复。我们报告了抗生素对生物膜形态的新的可量化影响,特别是生物膜流线。新设计连接了MBEC®分析设计,选择生物膜表型与原位成像需求。
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A microtiter peg lid with ziggurat geometry for medium-throughput antibiotic testing and in situ imaging of biofilms

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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来源期刊
Biofilm
Biofilm MICROBIOLOGY-
CiteScore
7.50
自引率
1.50%
发文量
30
审稿时长
57 days
期刊介绍:
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