title: Automated analysis of invadopodia dynamics in live cells.
abstract: Multiple cell types form specialized protein complexes that are used by the cell to actively degrade the surrounding extracellular matrix. These structures are called podosomes or invadopodia and collectively referred to as invadosomes. Due to their potential importance in both healthy physiology as well as in pathological conditions such as cancer, the characterization of these structures has been of increasing interest. Following early descriptions of invadopodia, assays were developed which labelled the matrix underneath metastatic cancer cells allowing for the assessment of invadopodia activity in motile cells. However, characterization of invadopodia using these methods has traditionally been done manually with time-consuming and potentially biased quantification methods, limiting the number of experiments and the quantity of data that can be analysed. We have developed a system to automate the segmentation, tracking and quantification of invadopodia in time-lapse fluorescence image sets at both the single invadopodia level and whole cell level. We rigorously tested the ability of the method to detect changes in invadopodia formation and dynamics through the use of well-characterized small molecule inhibitors, with known effects on invadopodia. Our results demonstrate the ability of this analysis method to quantify changes in invadopodia formation from live cell imaging data in a high throughput, automated manner.
authors: UNC/NCSU Joint Department of Biomedical Engineering, University of North Carolina at Chapel Hill , Chapel Hill, NC , USA.
authors: Matthew E
authors: ME
authors: Berginski
authors: Department of Cell Biology and Physiology, University of North Carolina at Chapel Hill , Chapel Hill, NC , USA.
authors: Sarah J
authors: SJ
authors: Creed
authors: UNC/NCSU Joint Department of Biomedical Engineering, University of North Carolina at Chapel Hill , Chapel Hill, NC , USA.
authors: Shelly
authors: S
authors: Cochran
authors: Department of Cell Biology and Physiology, University of North Carolina at Chapel Hill , Chapel Hill, NC , USA.
authors: David W
authors: DW
authors: Roadcap
authors: Department of Cell Biology and Physiology, University of North Carolina at Chapel Hill , Chapel Hill, NC , USA ; Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill , Chapel Hill, NC , USA ; Howard Hughes Medical Institute , Chevy Chase, MD , USA.
authors: James E
authors: JE
authors: Bear
authors: UNC/NCSU Joint Department of Biomedical Engineering, University of North Carolina at Chapel Hill , Chapel Hill, NC , USA ; Department of Computer Science, University of North Carolina at Chapel Hill , Chapel Hill, NC , USA ; Department of Pharmacology, University of North Carolina at Chapel Hill , Chapel Hill, NC , USA.
authors: Shawn M
authors: SM
authors: Gomez
keywords: Cancer; ECM degradation; Fluorescence microscopy; Image analysis; Invadopodia; Invasion; Live cell imaging; Metastasis; Podosomes