Phenotypic and MoA Analysis of Anti-Metastatic Molecules Using 3D Spheroid-Based Tumor Invasion Assays, Kinetic Microplate Detection, and Cellular Microscopy
Brad Larson
Wednesday, May 6, 2015
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Chapters
Introduction
Background
Outline
Synergy Neo
Cytation 5
Filter & Monochromator-based optics for optimal multi-mode detection
Built-in Inverted Fluorescence Microscope
Imaging & Microscopy: Z-Stacking, Z-Projection
Corning® Spheroid Microplate
Cell Culture Model Comparison
Spheroid Formation in Corning® Spheroid Microplates
3D Tumor Invasion Experimental Outline
3D Tumor Invasion Procedure
MDA-MB-231/Fibroblast Tumoroid Invasion
Total Invasion Analysis (using Brightfield Signal Mask)
Fluorescent Tracking of MDA-MB-231 GFP Invasion
Invasion Analysis (using GFP Signal Mask)
Z-Projection of Invading Structure
Determination of Cell Involvement in Total Invasion
EnSens® Protease Activity Detection Technology
Technology Integration into 3D Tumor Invasion Procedure
Quantification of MMP-14 Activity
Comparison of MMP-14 Activity to Total Invasion
Interruption of Invasion via MMP Activity Inhibition
Substrate Selectivity and Far Red Dye Signal Confirmation
Tag-Lite: Receptor Ligand Binding
SDF1-α-d2 Kinetic Characterization
Inhibitor Residence Time Assessment
Phenotypic Effect on 3D Tumoroid Invasion
Cellular Analysis of Tumoroid Invasion
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