Label-free Spore Counting of Bacterial Species

Traditional bacterial counting methods rely on time-consuming agar plate assays or fluorescent labeling that add steps and potential errors. Holotomography detects individual spores label-free using refractive index contrast, and the Bacterial Cell Counter software automates CFU quantification across large scanning areas. This note provides a complete workflow from sample preparation to data analysis, with validation against conventional counting methods.

What's inside

- Step-by-step protocol for label-free spore sample preparation and matrix-scanning image acquisition

- Automated CFU calculation using Bacterial Cell Counter with RI and size thresholds

- Concentration guidelines and detection limit table (10⁶–10⁹ spores/mL range)

- Applicability to vegetative bacteria (E. coli, Bacillus), fungal spores, and microbead counting

Keywords

bacterial spore counting CFU quantification label-free bacteria imaging holotomography Bacillus subtilis endospore detection colony forming unit refractive index tomography automated cell counting microbiology imaging high-throughput screening

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