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Isolating motile derivatives
- Streak each strain carefully at one point of a 0.7% TBAB agar plate.
- Incubate overnight at 37ºC without inverting the plates.
- Motile cells will swarm over the surface of the plate and should be picked from as far from the site of inoculation as possible. If the strain was non-motile there will be a contained streak with a few dendritic swarms emerging from the streak. Pick from the top of one of these swarms to recover a motile derivative.
(For GFP Microscopy)
- Grow cells overnight at 22°C in LB to 1 OD_600
- Pellet 1ml and resuspend in 0.1ml PBS
- Stain membranes by resuspension in 50μL of PBS containing 5μg/mL FM4-64 and incubate for 10min at room temperature.
- Observe samples by spotting 3μL of suspension on a glass microscope slide and immobilise with poly-L-lysine-treated glass coverslip.
- Capture images