Christian Doppler Laboratory for Molecular Food Analytics, Veterinärplatz 1, 1210 Vienna, Austria.
Appl Environ Microbiol. 2010 Apr;76(8):2600-6. doi: 10.1128/AEM.01506-09. Epub 2010 Feb 19.
The aim of this study was to observe growth of isolated single bacterial cells in the absence of growth factors and intercellular contact. In order to exclude stochastic uncertainties induced by dilution series, a new micromanipulation method was developed to ensure explicit results under visual control. This was performed with particular care for production of single prokaryotic cells and subsequent investigation of their autonomous growth. Over 450 single isolated Listeria monocytogenes and Salmonella enterica subsp. enterica serovar Typhimurium cells in lag, log, and stationary growth phases were investigated by this method, which included thoroughly washing the cells. The proportion of living cells within the initial cultures was compared to the proportion of positive samples after enrichment of the separated single cells. This resulted in P values of >/=0.05 using the chi-square test for statistical analysis, indicating no significant difference, and clearly demonstrates reproduction of isolated single bacterial cells without the need for growth factors or intercellular contact. Ease of handling of the apparatus and good performance of the cleaning procedures were achieved, as was validation of the method, demonstrating its suitability for routine laboratory use.
本研究旨在观察在缺乏生长因子和细胞间接触的情况下,单个细菌细胞的生长情况。为了排除稀释系列引起的随机不确定性,开发了一种新的微操作方法,以在可视化控制下确保明确的结果。特别注意生产单个原核细胞,并随后研究其自主生长。通过这种方法研究了超过 450 个处于潜伏期、对数期和稳定期的单个分离的李斯特菌和肠炎沙门氏菌亚种肠炎血清型 Typhimurium 细胞,包括彻底清洗细胞。通过对初始培养物中活细胞的比例与分离的单个细胞富集后的阳性样本比例进行比较,使用卡方检验进行统计分析,结果表明 P 值大于等于 0.05,无显著差异,明确证明了无需生长因子或细胞间接触即可复制分离的单个细菌细胞。该设备易于操作,清洗程序性能良好,方法得到验证,表明其适用于常规实验室使用。