Automated time-lapse microscopy and high-resolution tracking of cell migration.

Abstract:

:We describe a novel fully automated high-throughput time-lapse microscopy system and evaluate its performance for precisely tracking the motility of several glioma and osteoblastic cell lines. Use of this system revealed cell motility behavior not discernable with conventional techniques by collecting data (1) from closely spaced time points (minutes), (2) over long periods (hours to days), (3) from multiple areas of interest, (4) in parallel under several different experimental conditions. Quantitation of true individual and average cell velocity and path length was obtained with high spatial and temporal resolution in "scratch" or "wound healing" assays. This revealed unique motility dynamics of drug-treated and adhesion molecule-transfected cells and, thus, this is a considerable improvement over current methods of measurement and analysis. Several fluorescent vital labeling methods commonly used for end-point analyses (GFP expression, DiO lipophilic dye, and Qtracker nanocrystals) were found to be useful for time-lapse studies under specific conditions that are described. To illustrate one application, fluorescently labeled tumor cells were seeded onto cell monolayers expressing ectopic adhesion molecules, and this resulted in consistently reduced tumor cell migration velocities. These highly quantitative time-lapse analysis methods will promote the creation of new cell motility assays and increase the resolution and accuracy of existing assays.

journal_name

Cytotechnology

journal_title

Cytotechnology

authors

Fotos JS,Patel VP,Karin NJ,Temburni MK,Koh JT,Galileo DS

doi

10.1007/s10616-006-9006-7

subject

Has Abstract

pub_date

2006-05-01 00:00:00

pages

7-19

issue

1

eissn

0920-9069

issn

1573-0778

journal_volume

51

pub_type

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