Difference between revisions of "Huang: Agarose Slides for Imaging"
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Revision as of 12:55, 21 February 2009
This protocol allows live bacterial cell imaging. We create a small, nutrient rich gel pad and inject the bacteria on top.
- 500μg 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)
- Mix Agarose with buffer solution, microwave until boiling.
- Prepare one glass slide and position 2 thin cover slips on each end, to create 2 edges.
- Pipet 25 μl of the Agarose solution onto the middle of the glass slide, while making sure not to touch the coverslips.
- Quickly position the second glass slide on top of the molten gel drop.
- Leave to dry for a few minutes.
- Remove the top glass slide.
- Add 2-4 μl of the liquid culture onto the hardened gel drop.
- Leave to dry for a couple of minutes
- Add a cover slip on top, seal with nail polish at all 4 corners.
Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!