Real-time monitoring of single bacterium lysis and leakage events by chemiluminescence microscopy.

The small size of bacteria makes it difficult to study the biochemistry inside single cells. The amount of material inside is limited; therefore, an ultrasensitive method is required to interrogate single cells. Using a sensitive ICCD detector to record chemiluminescence (CL) from an optimized firefly luciferase-ATP bioluminescence reaction system, we report for the first time real-time imaging of lysis and leakage of single bacterium with 10-s temporal resolution. Movies are generated to visualize how the cell membrane was damaged by phage lysis, antibiotics attack, or dehydration, as well as the wall repair and cell recovery processes. The results show single-cell variations that are not obtainable from bulk measurements, confirming that CL microscopy of luciferase-expressing bacteria is a powerful tool for studying the fundamental biology of cells.