Dynamic staining of Bacillus endospores with Thioflavin T

Rapid detection and identification of endospores presents a range of complex challenges. Dynamic staining approach, developed in our lab, utilizes the time-course fluorescence enhancement of an amyloid-staining dye, Thioflavin T (ThT), after mixing with intact endospores. We examined the kinetics of staining Bacillus atrophaeus and Bacillus thuringiensis endospores, and the rates of staining were different for the two bacilli when intact endospores were treated with ThT. This finding demonstrates an avenue for attaining information about the sporulated bacterial species without lysing, germinating or other pretreatment steps.

[1]  Vicente Nuñez,et al.  Amyloid Histology Stain for Rapid Bacterial Endospore Imaging , 2011, Journal of Clinical Microbiology.

[2]  A. Ponce,et al.  Rapid Endospore Viability Assay of Clostridium sporogenes spores. , 2009, International journal of food microbiology.

[3]  Adam Driks,et al.  Morphogenesis of Bacillus Spore Surfaces , 2003, Journal of bacteriology.

[4]  Rong Wang,et al.  Fingerprinting species and strains of Bacilli spores by distinctive coat surface morphology. , 2007, Langmuir : the ACS journal of surfaces and colloids.

[5]  Terrance J Leighton,et al.  Architecture and high-resolution structure of Bacillus thuringiensis and Bacillus cereus spore coat surfaces. , 2005, Langmuir : the ACS journal of surfaces and colloids.

[6]  Guilford Jones,et al.  Nonlithographic fabrication of microfluidic devices. , 2006, Journal of the American Chemical Society.

[7]  H. Gray,et al.  Detection of bacterial spores with lanthanide-macrocycle binary complexes. , 2009, Journal of the American Chemical Society.

[8]  Duoduo Bao,et al.  Kinetics of bacterial fluorescence staining with 3,3'-diethylthiacyanine. , 2010, Langmuir : the ACS journal of surfaces and colloids.

[9]  Adrian Ponce,et al.  Bacterial spore detection by [Tb3+(macrocycle)(dipicolinate)] luminescence. , 2007, Journal of the American Chemical Society.

[10]  P. Eichenberger,et al.  The Bacillus subtilis spore coat protein interaction network , 2006, Molecular microbiology.

[11]  Leslie D. Knecht,et al.  Bacterial spores as platforms for bioanalytical and biomedical applications , 2011, Analytical and bioanalytical chemistry.

[12]  A. Malkin,et al.  In vitro high-resolution structural dynamics of single germinating bacterial spores , 2006, Proceedings of the National Academy of Sciences.