Nanovid tracking: a new automatic method for the study of mobility in living cells based on colloidal gold and video microscopy.
We describe a new automatic technique for the study of intracellular mobility. It is based on the visualization of colloidal gold particles by video-enhanced contrast light microscopy (nanometer video microscopy) combined with modern tracking algorithms and image processing hardware. The approach can be used for determining the complete statistics of saltatory motility of a large number of individual moving markers. Complete distributions of jump time, jump velocity, stop time, and orientation can be generated. We also show that this method allows one to study the characteristics of random motion in the cytoplasm of living cells or on cell membranes. The concept is illustrated by two studies. First we present the motility of colloidal gold in an in vitro system of microtubules and a protein extract containing a kinesin-like factor. The algorithm is thoroughly tested by manual tracking of the videotapes. The second study involves the motion of gold particles microinjected in the cytoplasm of PTK-2 cells. Here the results are compared to a study using the spreading of colloidal gold particles after microinjection.
Geerts
H
H
Department of Life Sciences, Janssen Pharmaceutica Research Laboratories, Beerse, Belgium.
De Brabander
M
M
Nuydens
R
R
Geuens
S
S
Moeremans
M
M
De Mey
J
J
Hollenbeck
P
P
D000818:Animals; D001322:Autoanalysis; D002460:Cell Line; D002465:Cell Movement; D006046:Gold; D008853:Microscopy / Q000379:methods; D014741:Video Recording
D006046:Gold