An agarose-in-plug bridge method to study chemotaxis in the Archaeon Halobacterium salinarum

Abstract

A simple agarose-in-plug bridge method was developed to study chemotaxis in the Archaeon Halobacterium salinarum. Preheated liquid agarose solution with chemoeffectors is pipetted in the middle of a microscope slide bridge, constructed by placing two plastic strips 16 mm apart. A coverslip is immediately placed over the agarose. The solidified agarose plug is completely encircled with the halobacterial cell suspension. Within a certain time concentrated halobacteria were seen as a ring at the edge of the agarose plug containing attractant amino acids and the control growth medium. Chemotaxis mutant Pho60 cells do not accumulate either around the attractants or around the growth medium. The kinetics of the ring formation can be readily videotaped or photographed using either phase-contrast or dark-field microscopy.