Project 2: Single Cell High Parameter Network Analysis by Phospho-Flow and Mass Tags
- Signaling systems can now be analyzed directly by Mass Cytometry, conventional flow cytometry, and Fluorescence Activated Cell Sorting. We have developed a series of methods for following multiple phosphoproteins in complex populations of primary cells.
- Up to 34 simultaneous parameters can be followed in single cells including multiple kinases, phosphoproteins, cell cycle, and other parameters allow for exacting resolution of cellular activation states.
- We are using these techniques to study B and T cell signaling, dendritic cell function, and other immune parameters by analysis of biochemical functions at the single cell level
- We currently have efforts in Pulmonary Arterial Hypertension, Acute Myelogenous Leukemia (adult and pediatric), Juvenile Monomyelocytic Leukemia, Follicular Lymphoma, Colon cancer, high throughput drug screening against kinases in complex cell populations, normal stem cell development, single cell kinetics and the cell cycle, so-called 'cancer stem cells', use of Raman spectroscopy for multiparametric cell imaging, among many other projects.
- A critical component of our work is the merging of complex biology assays with advanced computational methods for automated delineation of signaling networks (see our papers in Science, Cell, Nature Biotechnology, Cytometry A for details).
- Datasets and protocols from this work available at the Nolan Lab Resource.