Live-Cell Computed Tomography Instrumentation Development
(W.M. Keck Foundation Award, Deirdre Meldrum, PI)
In 2010 Dr. Meldrum received a $1M grant from the Keck Foundation to develop a 3D cell tomograph. This project was motivated by the work we have been pursuing under our CEGS funding, and our desire to develop additional tools to assess the functional state of individual cells.
This four-year project aims to build a novel, live-cell imaging instrument for basic and clinical science application. The 3D microscope we call the “cell CT scanner” will provide functional images, revealing the molecular mechanisms underlying important metabolic and disease processes. Cell CT is analogous to diagnostic radiology CT in that we will reconstruct 3D images with isotropic, submicron resolution from hundreds of projections acquired from many angles as the cell is rotated. A key component of the research is to determine the best method for rotating cells, which must be done extremely precisely without harming the cell. We will investigate one method that rotates an optically trapped cell in a microfluidic vortex and another that uses an asymmetrical infrared light beam. We will use fluorescent antibody probes and fusion-protein constructs specific to the proteins we are interested in to label cells for emission CT scanning. We will validate the technology studying cells from immortalized epithelial cell lines representative of various stages of esophageal cancer, and disaggregated from human biopsies spanning the same disease spectrum. Cell CT will enable for the first time rapid estimation of local protein concentrations in cellular compartments and microdomains, providing powerful insights concerning relationships between cell structure, function and disease.