Technical Reference #1999

1999.

Changes in Min oscillation pattern before and after cell birth Jennifer R. Juarez and William Margolin, University of Texas Medical School at Houston, Journal of Bacteriology, 192(1999), (2010)
Link To Paper

Keywords:
Min proteins; cytokinesis; E. coli; microscopy; Z ring

Materials & Methods:
Growth conditions and microscopy. A single colony was inoculated in Luria-Bertani (LB) medium with 50-100 μM IPTG and grown at 30°C or 37°C until mid-logarithmic phase. Three μl of culture was then placed on a cover glass on a glass-bottomed culture dish (Mattek Corp.) and overlaid with a thin strip of 1.5% LB agarose. Time lapse microscopy was done with a fully automated Olympus IX-71 outfitted with a 100X objective (N.A. 1.4) and filter wheels with exposure intervals from 1.5-2 seconds and exposure times of 0.5-0.8 seconds to 488 nm excitation light. The growth temperature on the slide was maintained at 32-33°C with a WeatherStation enclosure. Identical intervals and exposure times were used during any given time course. At these temperatures the GFP-MinD oscillation period was often as fast as 10 s (34) and cells grew significantly (0.5-1 μm) and sometimes divided during the time courses which were mostly 6-8 min in length

Microscopic Technique
Time lapse microscopy

Cell Type(s)
E. coli