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- Introduction and overview
- Nanomedical treatment at the single cell level requires evaluation at the single cell level
- For evaluation purposes, does structure reveal function?
- The difficulty of anything but simple functional assays
- The need for assays which at least show correlation to functional activity
- Quantitative single cell measurements of one or more proteins per cell by flow and image/confocal cytometry
- Cell surface measures of protein expression on live, single cells
- High-throughput flow cytometric screening of bioactive compounds
- Challenges of measuring protein expression inside fixed, single cells
- When location is important 2D or 3D imaging is required to get spatial location of proteins inside cells
- Quantitative multiparameter phospho-specific flow cytometry
- Attempts to measure "functional proteins" by detecting phosphorylation
- Example of phospho-specific, multiparameter flow cytometry
- Example of measuring single cell gene silencing by phospho-specific flow cytometry
- Quantitative measures of gene expression – the promises and the realities
- Is gene expression at the single cell level really possible?
- Is it even useful to measure a single gene's changes?
- Gene arrays of purified cell subpopulations
- RNA amplification techniques to attempt to perform single cell gene arrays
- Steven M. Chan, Janelle A. Olson, and Paul J. Utz. Single-Cell Analysis of siRNA-Mediated Gene Silencing Using Multiparameter Flow Cytometry. Cytometry Part A 69A:59–65 (2005).
- Peter O. Krutzik, Jonathan M. Irish, Garry P. Nolan and Omar D. Perez. Analysis of protein phosphorylation and cellular signaling events by flow cytometry: techniques and clinical applications. Clinical Immunology 110: 206– 221 (2004).
- Susan M. Young, Mark S. Curry, John T. Ransom, Juan A. Ballesteros, Eric R. Prossnitz, Larry A. Sklar and Bruce S. Edwards. High-Throughput Microfluidic Mixing and Multiparametric Cell Sorting for Bioactive Compound Screening. J Biomol Screening; 9; 103 – 111 (2004).
- Szaniszlo, P., Wang, N., Sinha, M., Reece, L.M., Van Hook, J.W., Luxon, B.A., Leary, J.F. "Getting the Right Cells to the Array: Gene Expression Microarray Analysis of Cell Mixtures and Sorted Cells" Cytometry 59A: 191-202 (2004).
- Szaniszlo, P. Gene Expression Microarray Analysis of Small, Purified Cell Subsets. University of Texas Medical Branch, Galveston, TX April, 2007 (mentor: Dr. Leary)
Researchers should cite this work as follows:
James Leary (2007), "BME 695N Lecture 16: Assessing drug efficacy at the single cell level," https://nanohub.org/resources/3478.
Biomedical Engineering, Room 1083