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11 © 2013 PerkinElmer
Dr. Matthias FasslerPerkinElmer Application Lab, Hamburg
Automated single cell tracking using the Operetta High Content Imaging System
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A549 non-small cell lung cancer cells were used to study chemokinesis (Caino et al., 2012)Chemokinesis = non-oriented increased migration after stimulationCells were seeded w/o serum onto collagen-coated 384 CellCarrier™ microplatesLive-cell imaging using a TCO-equipped Operetta® High Content Imaging SystemImaging every 5-10 mins for up to 6 hrs using a 10x objective and digital phase imaging
Digital phase imaging is non-toxic
A549 NSCLC Model
Constructed digital phase imageAcquisition of two brightfield images at suitable Z heights
HeLa cells, 20X hNA objective
upper plane
lower plane
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Object Tracking Workflow
Segment images Track objectsAcquire time series
Building block Track Objects of Harmony® Imaging and Analysis Software was used to tracks cells based on overlap between adjacent time points
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A549 migration is stimulated by EGF-treatment
Observed track length increases after EGF stimulation
A549 NSCLC Model
No EGF 100 ng/ml EGF
55 www.cellularimaging.com
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