Optimizing Bacillus subtilis spore isolation and quantifying spore harvest purity.
J Microbiol Methods
; 87(3): 325-9, 2011 Dec.
Article
em En
| MEDLINE
| ID: mdl-21989299
ABSTRACT
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 1σ) 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.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Esporos Bacterianos
/
Bacillus subtilis
/
Carga Bacteriana
/
Microscopia
Tipo de estudo:
Evaluation_studies
Idioma:
En
Revista:
J Microbiol Methods
Ano de publicação:
2011
Tipo de documento:
Article
País de afiliação:
Estados Unidos