The use of drip flow and rotating disk reactors for Staphylococcus aureus biofilm analysis.
The use of drip flow and rotating disk reactors for Staphylococcus aureus biofilm analysis.
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DOI：
10.3791/2470
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发表时间：
2010-12-27
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影响因子：
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通讯作者：
Boles, Blaise R
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作者：
Schwartz, Kelly;Stephenson, Rachel;Boles, Blaise R
自然界中的大多数微生物被认为是以表面相关群落的形式存在于生物膜中的。(1)细菌生物膜被包裹在基质中并附着在表面。(2)生物膜的形成和发展通常在实验室使用间歇系统(如微量滴定板)或流动系统(如流动细胞)进行研究。这些方法对于筛选突变和化学文库(微滴定板)(3)或用于可视化的生长生物膜(流动细胞)(4)是有用的。在这里，我们提出了在另外两种类型的流动系统生物膜中培养金黄色葡萄球菌的详细方案：滴流生物膜反应器和转盘生物膜反应器。滴流生物膜反应器是为研究在低剪切条件下生长的生物膜而设计的。(5)滴流反应器由四个平行的试验通道组成，每个通道能够容纳一张标准玻片大小的试片，或一段导管或支架。滴流反应器是微传感器监测、一般生物膜研究、生物膜冷冻切片样品、高生物量生产、医疗材料评估和留置医疗器械测试的理想选择。(6、7、8、9)旋转圆盘反应器由包含可拆卸优惠券凹槽的聚四氟乙烯圆盘组成。(10)可拆卸优惠券可以由任何可加工材料制成。旋转圆盘的底部包含一个条形磁铁，允许圆盘旋转以产生跨表面的液体表面剪切-冲洗优惠券。包含18张优惠券的整个圆盘被放置在1000毫升的玻璃侧臂反应堆容器中。当磁盘由磁力搅拌器旋转时，液体生长介质通过容器循环。将试片从反应堆容器中取出，然后刮取生物膜样本，以便进一步研究或进行显微镜成像。旋转盘式反应器设计用于实验室评估杀菌效果、去除生物膜和防污材料的性能。
Most microbes in nature are thought to exist as surface-associated communities in biofilms.(1) Bacterial biofilms are encased within a matrix and attached to a surface.(2) Biofilm formation and development are commonly studied in the laboratory using batch systems such as microtiter plates or flow systems, such as flow-cells. These methodologies are useful for screening mutant and chemical libraries (microtiter plates)(3) or growing biofilms for visualization (flow cells)(4). Here we present detailed protocols for growing Staphylococcus aureus in two additional types of flow system biofilms: the drip flow biofilm reactor and the rotating disk biofilm reactor. Drip flow biofilm reactors are designed for the study of biofilms grown under low shear conditions.(5) The drip flow reactor consists of four parallel test channels, each capable of holding one standard glass microscope slide sized coupon, or a length of catheter or stint. The drip flow reactor is ideal for microsensor monitoring, general biofilm studies, biofilm cryosectioning samples, high biomass production, medical material evaluations, and indwelling medical device testing.(6,7,8,9) The rotating disk reactor consists of a teflon disk containing recesses for removable coupons.(10) The removable coupons can by made from any machinable material. The bottom of the rotating disk contains a bar magnet to allow disk rotation to create liquid surface shear across surface-flush coupons. The entire disk containing 18 coupons is placed in a 1000 mL glass side-arm reactor vessel. A liquid growth media is circulated through the vessel while the disk is rotated by a magnetic stirrer. The coupons are removed from the reactor vessel and then scraped to collect the biofilm sample for further study or microscopy imaging. Rotating disc reactors are designed for laboratory evaluations of biocide efficacy, biofilm removal, and performance of anti-fouling materials.(9,11,12,13).