Project 5

Nanofluidic Devices for Rapid Single Cell Analysis
of Protein Expression

J. Michael Ramsey, Ph.D.,

Minnie N. Goldby Distinguished Professor of Chemistry, Project PI

This project is founded on the advanced micro- and nano-fluidics technology developed in the Ramsey laboratory at UNC and the Walker laboratory at North Carolina State University. A major thrust of the project is development of a novel micro/nano-fluidics cell analyzer/sorter to provide rapid analysis of normal and cancerous lymphocytes from blood and bone marrow samples of normal and tumor bearing mice and eventually from cancer patients. Analysis of other cell populations will also be pursued. Cell size and antibodies to cell surface markers will provide avenues to efficient sorting.

The sorted cells will then be analyzed for various growth-related parameters such as the state of activation of key kinases and GTPases; this will utilize the conformation-sensitive probes uniquely developed by the laboratory of Dr. Klaus Hahn of UNC Pharmacology.

Functionalized nanopores will be tested as detectors for specific proteins from lymphocytes so as to provide previously inaccessible lineage-specific ’signatures’ (for example, for regulatory T-cells).

Project 5 includes the following goals:

• Construction of the nanofluidics flow cytometer and its testing using well-defined cell types.
• Integration of a sample processing unit to allow clinicians access to near real-time diagnostic information.
• Design and fabrication of nanopore based sensors for cell-type specific protein signatures.
• Use of molecular biosensors in conjunction with the cytometer to analyze signaling events in individual cells.

Project 5 will make extensive use of the Animal Models Core and the Combinatorial Library Core. This project aims to develop both technological advances, such as the speed and precision of the sorter, and biological advances such as examination of signaling events in individual tumor cells.

Links:

Professor Ramsey's homepage