Morita Masamune, Katoh Kaoru, Noda Naohiro
Biomedical Research Institute National Institute of Advanced Industrial Science and Technology (AIST), Center 6 1-1-1 Higashi Tsukuba Ibaraki 305-8566 Japan.
ChemistryOpen. 2018 Aug 17;7(11):845-849. doi: 10.1002/open.201800126. eCollection 2018 Nov.
Bacterial cultivation techniques are classic, basic, and common processes used to characterize the physiological activity of bacteria in their environment. Owing to recent advances in bacterial cultivation techniques, the physiological activity of bacteria can be elucidated at the single-cell culture level. Here, we report a novel method to monitor the real-time activity of bacterial growth at the single-cell level inside giant unilamellar vesicles (GUVs). This method consists of two steps: 1) encapsulation of single bacteria in 1-33 pL scale GUVs and 2) immobilization of the GUVs on a planar lipid bilayer membrane on a glass surface. We directly observed single cells actively growing to a great number of cells inside GUVs. GUVs also protected the bacteria from external antibiotic compounds during prolonged cultivation for more than 24 h. This approach can be applied widely in the fields of biochemistry, biotechnology, microbiology, and synthetic biology.
细菌培养技术是用于表征细菌在其环境中的生理活性的经典、基础且常见的过程。由于细菌培养技术的最新进展，细菌的生理活性可以在单细胞培养水平上得到阐明。在此，我们报告一种在巨型单层囊泡（GUV）内单细胞水平监测细菌生长实时活性的新方法。该方法包括两个步骤：1）将单个细菌封装在1 - 33皮升规模的GUV中，以及2）将GUV固定在玻璃表面的平面脂质双分子层膜上。我们直接观察到单个细胞在GUV内积极生长为大量细胞。在超过24小时的长时间培养过程中，GUV还保护细菌免受外部抗生素化合物的影响。这种方法可广泛应用于生物化学、生物技术、微生物学和合成生物学领域。
