Assembly of minicolony development video from still images:
1) Cells are inoculated in microtiter wells as described in basic protocol.
2) 14 hrs after inoculation, a Nikon TEM inverted microscope was focused on a single minicolony with a 20X objective. It is much easier to focus reliably on a small cluster of cells in this system than on a single cell.
3) Images are captured using a standard digital camera attached to the microscope and also connected to a computer. We use a Canon Powershot G5 digital camera controlled by GBTimelapse software (Granite Bay Software) to capture images every 10 min for 5-10 days. Video was created from this series of images in Adobe® Premiere Pro CS4 and After Effects CS4. After the video was assembled, variation in luminescence between frames was reduced using VirtualDub (1.3).
