Huang: Agarose Slides for Imaging

Overview
This protocol allows live bacterial cell imaging. We create a small, nutrient rich gel pad and inject the bacteria on top.

Materials

 * 500mg Agarose
 * 100 ml PBS buffer/bacterial media
 * 2 Glass microscope slides
 * 4 Glass cover slips
 * Nail Polish
 * Bacterial liquid culture (at least 10 μl, ideally in exponential phase)

Procedure

 * 1) Mix Agarose with buffer solution, microwave until boiling.
 * 2) Prepare one glass slide and position 2 thin cover slips on each end, to create 2 edges.
 * 3) Pipet 25 μl of the Agarose solution onto the middle of the glass slide, while making sure not to touch the coverslips.
 * 4) Quickly position the second glass slide on top of the molten gel drop.
 * 5) Leave to dry for a few minutes.
 * 6) Remove the top glass slide.
 * 7) Add 2-4 μl of the liquid culture onto the hardened gel drop.
 * 8) Leave to dry for a couple of minutes
 * 9) Add a cover slip on top, seal with nail polish at all 4 corners.
 * 10) You are now ready to image some cells!

Contact

 * Carolina Tropini