Single-Cell Analysis

A review of current NIH funding (performed by NIH Sales*

^*Data provided by NIH Sales, Inc., 2 Snowmound Ct., Rockville, MD 20850, USA. Phone: 301-279-7175. www.nihsales.net

CEGS: Microscale Life Sciences Center

5P50HG002360-08, National Human Genome Research Institute

Deirdre R. Meldrum (Arizona State University-Tempe Campus, Tempe, AZ)

$3,497,560

Goal: To link preexisting cell state to cell fate in the disease process in heterogeneous cell populations at the individual cell level, by developing microscale technology for high-throughput genomic-level and multi-parameter single-cell analysis. New technology will be integrated into Living Cell Analysis cassette system, including 1) automated systems for cell placement, off-chip device interconnects, and high throughput data analysis; 2) optical and electronic sensors, dyes, and nanowires; and 3) micromodules for single-cell qRT-PCR, LATE-PCR for LOH including single-cell pyrosequencing, on-chip single-cell proteomics, and single-cell transcriptomics using barcoded nanobeads.

Optics Instrumentation for Advanced Ophthalmic Imaging

5R01EY014375-05, National Eye Institute

David R. Williams (University Of Rochester, Rochester, NY)

$1,942,063

Goal: 1) To design and construct instruments for noninvasive imaging of the mammalian retina with 3-D resolution capable of resolving single cells in vivo. 2) To explore the value of this technology through application to human retinal disease and retinal surgery. Instruments will combine adaptive optics with confocal microscopy.

Alliance for Cellular Signaling

5U54GM062114-08, National Institute of General Medical Sciences

Alfred G. Gilman (University of Texas Southwest Medical Center, Dallas, TX)

$1,414,235

Goal: To understand cellular communication as a network of modules and components, to measure interactions between modules, to apply and develop tools to measure module “state” and information flow between modules, and to collect data to support quantitative models of specific modules.

Raman Flow Cytometry for Diagnostics and Drug Discovery

5R01EB003824-04, National Institute of Biomedical Imaging and Bioengineering

John P. Nolan (La Jolla Bioengineering Institute, La Jolla, CA)

$1,171,157

Goal: To develop a new analytical platform for high-throughput screening and selection based on Raman Flow Cytometry, including instrumentation, optically encoded polymer resins for chemical synthesis and screening, and nanostructured materials with unique optically properties for sensitive reporting and encoding. The new technology will perform Raman spectroscopy on single particles in flow to enable new applications in sensitive multiplexed detection, drug discovery, and diagnostics.

Radiological Research Accelerator Facility (RARAF)

5P41EB002033-12, National Institute of Biomedical Imaging and Bioengineering

David J. Brenner (Columbia University Health Sciences, New York, NY)

$888,877

Goal: To improve accuracy, enhance imaging capabilities, extend the charged-particle microbeam to X rays; and to develop a simple stand-alone microbeam that can be used without physics support (which might be exported to other labs). Objectives include a custom multi-photon imaging system to allow time-lapse fluorescent imaging of early (scale of seconds to minutes) radiation events in individual live cells, a new imaging system based on quantitative phase microscopy to image live cells and cell nuclei without cytotoxic stains, and a system for single-cell microsensors to monitor the flux of molecules such as oxygen and No.

Signal Transduction by Tyrosine Phosphorylation

5R01CA082683-09, National Cancer Institute

Tony R. Hunter (Salk Institute for Biological Studies, La Jolla, CA)

$656,267

Goal: To study how signals are transmitted to the action cytoskeleton, controlling local assembly and remodeling in response to tyrosine kinase mediated signaling. The research will develop genetically encoded FRET-based kinase activity reporters to study spatiotemporal patterns of protein kinase activity in single living cells, specifically along the pathway used by c-Abl tyrosine kinase to regulate assembly of the action cytoskeleton and promote formation of surface filopodia in cells spreading on ECM.

Single-Molecule Fluorophores for Cellular Imaging

5P20HG003638-04, National Human Genome Research Institute

William E. Moerner (Stanford University, Stanford, CA)

$648,597

Goal: In a high-risk/high-potential-payoff program, to design, develop, and apply unique fluorophores to explore protein localization in cells, down to the single-molecule level. The research involves 1) design, synthesis, characterization, and optimization of a new class of fluorescent tags for biological imaging, 2) development of cellular targeting technologies for these new fluorophores to determine location and co-location of proteins, and 3) demonstration of methods for detection of coordinated protein location (or mislocation in mutant strains) and gene expression to explore the regulatory function of spatial positioning in bacteria.

^*Data provided by NIH Sales, Inc., 2 Snowmound Ct., Rockville, MD 20850, USA. Phone: 301-279-7175. www.nihsales.net