Cell Enumeration of Leptospira by Flow Cytometry

Rapid and reliable enumeration of Leptospira spp., the causative agent of leptospirosis, represents a technical challenge because leptospires are thin, highly motile, and slow-growing bacteria. The current gold standard for cell enumeration is the use of a Petroff-Hausser counting chamber and a dark-field microscope, but this method remains time-consuming and lacks reproducibility. New alternative techniques are then of great interest. Here we describe the protocol for counting leptospires by flow cytometry. This method is rapid, reproducible, sensitive, and hence suitable to become a new standard to enumerate Leptospira spp.