The monocytes were imaged with epifluorescence with an Olympus 60X N.A. 1.4 objective at the rate of 8 frames per second with a Photometrics PXL camera. The first timepoint is approximately 3 minutes after the cells were plated on glass. Notice that the macrophage has already begun to spread dramatically. It has also sent out very dynamic processes to touch neighboring cells and to assess whether to phagocytose them. The graph at the left shows the increased spreading of the monocyte as it attempts to ingest the glass coverslip.
Small spikes show quick variations such as long protrusions which quickly retract.
These data were generated from the movie by an automated process outlined below. The diagram at the left shows the outlines of the cell at all time points. The numbers at the center show the location of the centroid of the cell and the path traveled. The quantitation of each frame in the time lapse movie was achieved using the script at the left. Before running the script, the user chooses the threshold values that the cell needs to be extracted from other features in the image.