A method for growing a biofilm under low shear at the air-liquid interface using the drip flow biofilm reactor
A method for growing a biofilm under low shear at the air-liquid interface using the drip flow biofilm reactor
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DOI：
10.1038/nprot.2009.59
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发表时间：
2009-01-01
期刊：
影响因子：
14.8
通讯作者：
Stewart, Philip S.
中科院分区：
文献类型：
--
作者：
Goeres, Darla M.;Hamilton, Martin A.;Stewart, Philip S.
该方案描述了如何使用滴流反应器(DFR)在靠近气液界面的低流体剪切条件下生长铜绿假单胞菌生物膜。DFR可以模拟食品加工传送带、导管、患有囊性纤维化的肺部和口腔等环境。生物膜是通过间歇运行6h的方式建立的，当反应器在连续的营养物质流中运行48h时，成熟的生物膜形成。在连续流动过程中，当介质滴到以10度角设置的表面上时，生物膜经历低剪切。在54h结束时，通过去除反应器通道中的菌球，漂洗菌球以去除浮游细胞，刮除菌球表面的生物膜，解聚团块，然后稀释和培养活细胞计数来量化生物膜的积累。整个过程需要13小时的活动时间，分布在5天内。
This protocol describes how to grow a Pseudomonas aeruginosa biofilm under low fluid shear close to the air-liquid interface using the drip flow reactor (DFR). The DFR can model environments such as food-processing conveyor belts, catheters, lungs with cystic fibrosis and the oral cavity. The biofilm is established by operating the reactor in batch mode for 6 h. A mature biofilm forms as the reactor operates for an additional 48 h with a continuous flow of nutrients. During continuous flow, the biofilm experiences a low shear as the media drips onto a surface set at a 10 degrees angle. At the end of 54 h, biofilm accumulation is quantified by removing coupons from the reactor channels, rinsing the coupons to remove planktonic cells, scraping the biofilm from the coupon surface, disaggregating the clumps, then diluting and plating for viable cell enumeration. The entire procedure takes 13 h of active time that is distributed over 5 d.