Application of Bioimage Informatics to Quantification of Focal Adhesions and Invadopodia.

摘要

The development of the ability to fluorescently label functional proteins and visualize their subcellular localization using microscopy in living cells, has made it possible to study a wide range of single cell phenomena. To understand the results of imaging assays, cell biologists have used manual methods for determining the quantitative properties of the cellular structures visualized fluorescent microscopy. As the quantity and complexity of the images that can be collected using fluorescence microscopy has increased, a new subfield of Bioinformatics has developed, named Bioimage Informatics, which specializes in adapting and developing new methods to quantify the image sets resulting from biological assays.;In this thesis, I describe the application and development of Bioimage Informatic methods to the analysis of Focal Adhesions and Invadopodia. Focal Adhesions are subcellular protein complexes, whose role include acting as the points of contact for cellular motility and sensing the outside environment. Focal Adhesions have traditionally been analyzed using manual methods, which has limited the number of Focal Adhesions that could be analyzed and the depth of properties that could be collected. I have developed a set of methods which can identify, track and quantify Focal Adhesion properties from live cell image sets. This Focal Adhesion analysis framework has been expanded to include spatial and global methods for describing Focal Adhesion morphology. I have also developed a system for quantifying Invadopodia properties. Invadopodia are subcellular protein complexes present in metastatic cancer cells, which actively degrade the extracellular matrix, allowing migration of cancer cells away from primary tumors. This analysis system has two parts, one which can follow single Invadopodia and assess their properties and a complementary component which assesses degradation behavior in cell populations.