Image-based cell-resolved screening assays in flow

Cytometry A. 2015 Jun;87(6):541-8. doi: 10.1002/cyto.a.22609. Epub 2014 Dec 16.

Abstract

A parallel microfluidic cytometer (PMC) is based on a one-dimensional (1D) scanning detector, a parallel array of flow channels, and new multiparameter analysis algorithms that operate on low-pixel-count 1D images. In this article, we explore a series of image-based live- and fixed-cell screening assays, including two NF-kB nuclear translocations and T-cell capping. We then develop a new multiparametric linear weighted classifier that achieves a Z' factor sufficient for scaled pharmaceutical discovery with Jurkat cells in suspension. We conclude that the PMC should have the throughput and statistical power to permit a new capability for image-based high-sample-number pharmaceutical screening with suspension samples.

Keywords: HCA; high-content; high-throughput; imaging; live-cell assay; microfluidic; receptor capping; screening.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Algorithms
  • Animals
  • CHO Cells
  • Carcinoma, Hepatocellular / drug therapy
  • Cell Line, Tumor
  • Computational Biology / methods
  • Cricetulus
  • Drug Discovery / methods*
  • Flow Cytometry / instrumentation
  • Flow Cytometry / methods*
  • Green Fluorescent Proteins
  • High-Throughput Screening Assays / methods
  • Humans
  • Image Processing, Computer-Assisted / methods*
  • Jurkat Cells
  • Liver Neoplasms / drug therapy
  • Microfluidic Analytical Techniques / instrumentation
  • Microfluidic Analytical Techniques / methods*
  • NF-kappa B / physiology
  • Systems Biology / methods
  • T-Lymphocytes / physiology

Substances

  • NF-kappa B
  • Green Fluorescent Proteins