Preparation of Vibrio parahaemolyticus Swarmer Cells for Imaging

Take a dish containing a colony of Vibrio parahaemolyticus bacteria.

The bacteria exhibit two forms: short rod-shaped swimmer cells with a single polar flagellum located at the colony center and elongated swarmer cells with lateral flagella that migrate outward.

The cells are engineered to express a fluorophore-tagged protein, which localizes unipolarly in swimmer and bipolarly in swarmer cells, aiding in their identification.

Next, pour molten agarose between tape guides on a microscope slide and press it with a second slide to spread evenly.

Allow it to solidify, trim the excess, and remove the top slide to expose the agarose pad.

Cut a section from the colony edge and invert it onto the pad.

Allow the cells to adhere and remove the section.

Place a coverslip to prepare the cells for imaging.

Elongated cells with bipolarly distributed fluorescent protein indicate the presence of swarmer cells.

The first step in this procedure is to prepare agarose slides for microscopy.

Add one gram of agarose to 100 milliliters of a 20% PBS and a 10% LB broth solution. Carefully boil the solution in the microwave to dissolve the agarose, and then let it cool down to 65 to 70 degrees Celsius while mixing w