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BME 695N Lecture 16: Assessing drug efficacy at the single cell level

By James Leary

Purdue University

Published on

Abstract

Outline:
  1. Introduction and overview
    1. Nanomedical treatment at the single cell level requires evaluation at the single cell level
    2. For evaluation purposes, does structure reveal function?
    3. The difficulty of anything but simple functional assays
    4. The need for assays which at least show correlation to functional activity
  2. Quantitative single cell measurements of one or more proteins per cell by flow and image/confocal cytometry
    1. Cell surface measures of protein expression on live, single cells
    2. High-throughput flow cytometric screening of bioactive compounds
    3. Challenges of measuring protein expression inside fixed, single cells
    4. When location is important 2D or 3D imaging is required to get spatial location of proteins inside cells
  3. Quantitative multiparameter phospho-specific flow cytometry
    1. Attempts to measure "functional proteins" by detecting phosphorylation
    2. Example of phospho-specific, multiparameter flow cytometry
    3. Example of measuring single cell gene silencing by phospho-specific flow cytometry
  4. Quantitative measures of gene expression – the promises and the realities
    1. Is gene expression at the single cell level really possible?
    2. Is it even useful to measure a single gene's changes?
    3. Gene arrays of purified cell subpopulations
    4. RNA amplification techniques to attempt to perform single cell gene arrays

References

Outline:
  • 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)

Cite this work

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.

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Time

Location

Biomedical Engineering, Room 1083

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