Optimizing Bacillus subtilis spore isolation and quantifying spore harvest purity

被引:21
|
作者
Harrold, Zoe R. [1 ]
Hertel, Mikaela R. [1 ]
Gorman-Lewis, Drew [1 ]
机构
[1] Univ Washington, Earth & Space Sci Dept, Seattle, WA 98195 USA
基金
美国国家科学基金会;
关键词
Bacillus subtilis; Endospore; Isolation; Purification; Separation; MARINE BACILLUS; BACTERIAL SPORES; ENDOSPORES; OXIDATION; COATS;
D O I
10.1016/j.mimet.2011.09.014
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Investigating the biochemistry, resilience and environmental interactions of bacterial endospores often requires a pure endospore biomass free of vegetative cells. Numerous endospore isolation methods, however, neglect to quantify the purity of the final endospore biomass. To ensure low vegetative cell contamination we developed a quality control technique that enables rapid quantification of endospore harvest purity. This method quantifies spore purity using bright-field and fluorescence microscopy imaging in conjunction with automated cell counting software. We applied this method to Bacillus subtilis endospore harvests isolated using a two-phase separation method that utilizes mild chemicals. The average spore purity of twenty-two harvests was 88 +/- 11% (error is 1 sigma) with a median value of 93%. A spearman coefficient of 0.97 correlating automated and manual bacterial counts confirms the accuracy of software generated data. (C) 2011 Elsevier B.V. All rights reserved.
引用
收藏
页码:325 / 329
页数:5
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