Duran Clelia, Zhang Shiyi, Yang Chongyang, Falco Maria Lorena, Cravo-Laureau Cristiana, Suzuki-Minakuchi Chiho, Nojiri Hideaki, Duran Robert, Sassa Fumihiro
Universite de Pau et des Pays de l'Adour, E2S UPPA, CNRS, IPREM, Pau, France.
Graduate School of Information Science and Electrical Engineering, Kyushu University, Fukuoka, Japan.
Front Microbiol. 2022 Dec 16;13:1031439. doi: 10.3389/fmicb.2022.1031439. eCollection 2022.
In order to exploit the microbes present in the environment for their beneficial resources, effective selection and isolation of microbes from environmental samples is essential. In this study, we fabricated a gel-filled microwell array device using resin for microbial culture. The device has an integrated sealing mechanism that enables high-density isolation based on the culture of microorganisms; the device is easily manageable, facilitating observation using bright-field microscopy. This low-cost device made from polymethyl methacrylate (PMMA)/polyethylene terephthalate (PET) has 900 microwells (600 μm × 600 μm × 700 μm) filled with a microbial culture gel medium in glass slide-sized plates. It also has grooves for maintaining the moisture content in the micro-gel. The partition wall between the wells has a highly hydrophobic coating to inhibit microbial migration to neighboring wells and to prevent exchange of liquid substances. After being hermetically sealed, the device can maintain moisture in the agarose gels for 7 days. In the bacterial culture experiment using this device, environmental bacteria were isolated and cultured in individual wells after 3 days. Moreover, the isolated bacteria were then picked up from wells and re-cultured. This device is effective for the first screening of microorganisms from marine environmental samples.
为了利用环境中微生物的有益资源，从环境样本中有效筛选和分离微生物至关重要。在本研究中，我们使用树脂制造了一种用于微生物培养的凝胶填充微孔阵列装置。该装置具有集成密封机制，能够基于微生物培养进行高密度分离；该装置易于操作，便于使用明场显微镜进行观察。这种由聚甲基丙烯酸甲酯（PMMA）/聚对苯二甲酸乙二酯（PET）制成的低成本装置在玻片大小的平板中有900个微孔（600μm×600μm×700μm），填充有微生物培养凝胶培养基。它还有用于保持微凝胶中水分含量的凹槽。孔之间的隔壁具有高度疏水涂层，以抑制微生物迁移到相邻孔并防止液体物质交换。密封后，该装置可在琼脂糖凝胶中保持水分7天。在使用该装置的细菌培养实验中，3天后环境细菌在各个孔中被分离和培养。此外，然后从孔中取出分离的细菌并进行再培养。该装置对于从海洋环境样本中首次筛选微生物是有效的。
