了解抗生素耐药性的机制对于开发新的疗法至关重要。用于测试细菌的传统方法通常在其效率和可重用性方面受到限制。可以使用双重乳液以高通量研究单细菌细胞,尽管缺乏对油壳渗透性的控制和对液滴内部的有限访问存在严重的缺点。这里,介绍了一种研究细菌包封双乳液模板巨型单层囊泡(GUV)的简单策略。这种微流体方法用于同时将细菌装载在合成的GUV内,并用成孔肽蜂毒素渗透其膜。这使得抗生素递送或新鲜培养基流入GUV腔用于高度平行培养和抗微生物效力测试。基于聚合物的GUV被证明是有效的培养和分析微血管,因为微流体允许容易选择和封装的细菌和快速修改培养条件的抗生素开发。Further,证明了一种用于高通量筛选的GUV内生物膜原位谱分析的方法。可以想象,配备生物孔的合成GUV可以作为在生物膜形成过程中高通量筛选细菌菌落相互作用和研究抗生素对生物膜的影响的基础。
Understanding the mechanisms of antibiotic resistance is critical for the development of new therapeutics. Traditional methods for testing bacteria are often limited in their efficiency and reusability. Single bacterial cells can be studied at high throughput using double emulsions, although the lack of control over the oil shell permeability and limited access to the droplet interior present serious drawbacks. Here, a straightforward strategy for studying bacteria-encapsulating double emulsion-templated giant unilamellar vesicles (
GUVs) is introduced. This microfluidic approach serves to simultaneously load bacteria inside synthetic
GUVs and to permeabilize their membrane with the pore-forming peptide melittin. This enables antibiotic delivery or the influx of fresh medium into the GUV lumen for highly parallel cultivation and antimicrobial efficacy testing. Polymer-based
GUVs proved to be efficient culture and analysis microvessels, as microfluidics allow easy selection and encapsulation of bacteria and rapid modification of culture conditions for antibiotic development. Further, a method for in situ profiling of biofilms within
GUVs for high-throughput screening is demonstrated. Conceivably, synthetic
GUVs equipped with biopores can serve as a foundation for the high-throughput screening of bacterial colony interactions during biofilm formation and for investigating the effect of antibiotics on biofilms.