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Updated: May 14, 2026

Improvement of Bacillus subtilis Spore Enumeration and Label Analysis in Flow Cytometry
Published on: June 30, 2023
Dynamic staining of Bacillus endospores with Thioflavin T
Srigokul Upadhyayula1, Samuel Lam, Alice Ha
1Department of Biochemistry, Department of Bioengineering and Center for Bioengineering Research, University of California, Riverside, CA 92521, USA. supad003@ucr.edu
Abstract:
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.
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The simplest type of preparation is the wet mount, in which the specimen is placed in a drop of liquid on the slide. A liquid specimen can be directly deposited on the slide using a dropper. Solid specimens, such as skin scraping, can be placed on the slide before adding a drop of liquid to prepare the wet mount. Sometimes the liquid is simply water, but stains are often added...
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