Significantly improved precision of cell migration analysis in time-lapse video microscopy through use of a fully automated tracking system

Table 1 Cell speed variability caused by imprecise centroid selection

Cell type	σ in μm	original MD	AMD in μm/min	%	AMD (smoothed)	%
sc	3	0.181	0.423	234	0.125	69
sc	4.5		0.578	320	0.151	84
sc	6		0.747	414	0.181	100
sc	7.5		0.921	510	0.218	121
mc	3	0.622	0.723	116	0.465	75
mc	4.5		0.829	133	0.473	76
mc	6		0.956	154	0.484	78
mc	7.5		1.099	177	0.499	80
fc	3	1.787	1.821	102	1.589	89
fc	4.5		1.860	104	1.591	89
fc	6		1.923	108	1.597	89
fc	7.5		1.985	111	1.598	89

Three cell types where tested (slow cell (sc), medium fast cell (mc), fast cell (fc)). The centroid positions where artificially varied within a standard deviation (σ, both axes) of 3 to 7.5 μm around the real centroid and the average mean displacement (AMD) computed for each set of varied tracks (200 tracks per setting). Variation of centroid positions resulted in overestimation of cell speeds, which was most pronounced for the slowest cell. Smoothing of "noisy" cell tracks by a centered moving average filter (window size 5) tended to underestimate MD values to varying degrees.

ISSN: 2661-8850