Using 3D Super-Resolution Microscopy to Probe Breast Cancer Stem Cells and Their Microenvironment
Final rept. 15 Feb 2011-14 Feb 2014
CALIFORNIA UNIV BERKELEY
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We report the final achievements of our project. These achievements include the successful building of a super-resolution microscope that is capable of performing 2D and 3D Stochastic Optical Reconstruction Microscopy STORM and Photoactivated Localization Microscopy PALM, and a quantitative-phase imaging QPI system that enables the label-free imaging of cells over a period of days. With STORM imaging, we imaged the spatial organization or clustering of the CCR7 receptor on the surface of MCF-7 breast cancer cells that had been cultured on gelatin substrates. This imaging was the first of its kind for breast cancer. We also small controllable microenvironments, in which we can simultaneously culture small groups of cells and monitor their response to external perturbation. We subsequently examined the effects of microenvironments on breast cancer by creating arrays of polydimethlysiloxane PDMS microposts of different stiffness and sizes and seeded them with MCF-7 cells. We manipulated our micropost microenvironments to study the effects of microenvironment on cell lineage, namely the basal-luminal differentiation of MCF-7s. Finally, with QPI, we investigated the real-time response of breastcancer cells to different microenvironmental cues. We thus have begun to learn how protein density, growth rate, morphology, motility, and phenotype of these cells change and evolve in response to their microenvironment.
- Medicine and Medical Research