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See references below for related reading.
12.1 Introduction to measures of efficacy for nanomedicine
12.1.1 for evaluation purposes, does structure/size reveal function?
12.1.2 nanomedical treatment at the single cell level requires evaluation at the single cell level
12.1.3 the difficulty of anything but simple functional assays (e.g. phosphorylated “functional” proteins)
12.1.4 the need for assays which at least show correlation to functional activity
12.2 Quantitative single cell measurements of one or more proteins per cell by flow and image/confocal cytometry
12.2.1 cell surface measures of protein expression on live, single cells
12.2.2 high-throughput flow cytometric screening of bioactive compounds
12.2.3 challenges of measuring protein expression inside fixed, single cells
12.2.4 when location is important 2D or 3D imaging is required to get spatial location of proteins inside cells (“locational proteomics” at the single-cell level)
12.3 Quantitative multiparameter phospho-specific flow/image cytometry as a single-cell,structural-functional measurement
12.3.1 attempts to measure "functional proteins" by detecting phosphorylation
12.3.2 example of phospho-specific, multiparameter flow cytometry
12.3.3 example of measuring single cell gene silencing by phospho-specific flow cytometry
12.4 Quantitative measures of gene expression – the promises and the realities
12.4.1 is gene expression at the single cell level really possible?
12.4.2 is it even useful to measure a single gene's changes?
12.4.3 gene arrays of purified cell subpopulations
12.4.4 RNA amplification techniques to attempt to perform single cell gene arrays
- Chan, S.M., Olson, J.A., Utz, P.J. "Single-Cell Analysis of siRNA-Mediated Gene Silencing Using Multiparameter Flow Cytometry". Cytometry Part A 69A:59–65 (2005).
- Krutzik, P.O., Irish, J.M., Nolan, G.P., Perez, O.D. "Analysis of protein phosphorylation and cellularsignaling events by flow cytometry: techniques and clinical applications". Clinical Immunology 110:206– 221 (2004). (Full text available at nih.gov)
- 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 (2011), "BME 695L Lecture 12: Assessing Drug Efficacy and Nanotoxicity at the Single Cell Level," https://nanohub.org/resources/12392.
1083 BME, Purdue University, West Lafayette, IN