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Journal of Biomolecular Screening
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High-Throughput Microfluidic Mixing and Multiparametric Cell Sorting for Bioactive Compound Screening

Susan M. Young

Cytometry, Cancer Research and Treatment Center, University of New Mexico Health Sciences Center, Albuquerque

Mark S. Curry

Cytometry, Cancer Research and Treatment Center, University of New Mexico Health Sciences Center, Albuquerque

John T. Ransom

Novasite, Inc., 11095 Flintkote Avenue, San Diego, CA

Juan A. Ballesteros

Novasite, Inc., 11095 Flintkote Avenue, San Diego, CA

Eric R. Prossnitz

Cytometry, Cancer Research and Treatment Center, University of New Mexico Health Sciences Center, Albuquerque

Larry A. Sklar

Cytometry, Cancer Research and Treatment Center, University of New Mexico Health Sciences Center, Albuquerque

Bruce S. Edwards

Cytometry, Cancer Research and Treatment Center, University of New Mexico Health Sciences Center, Albuquerque bedwards{at}salud.unm.edu

HyperCyt®, an automated sample handling system for flow cytometry that uses air bubbles to separate samples sequentially introduced from multiwell plates by an autosampler. In a previously documented HyperCyt® configuration, air bubble separated compounds in one sample line and a continuous stream of cells in another are mixed in-line for serial flow cytometric cell response analysis. To expand capabilities for high-throughput bioactive compound screening, the authors investigated using this system configuration in combination with automated cell sorting. Peptide ligands were sampled from a 96-well plate, mixed in-line with fluo-4-loaded, formyl peptide receptor-transfected U937 cells, and screened at a rate of 3 peptide reactions per minute with ~ 10,000 cells analyzed per reaction. Cell Ca 2+ responses were detected to as little as 10-11 Mpeptide with no detectable carryover between samples at up to 10-7 M peptide. After expansion in culture, cells sort-purified from the 10% highest responders exhibited enhanced sensitivity and more sustained responses to peptide. Thus, a highly responsive cell subset was isolated under high-throughput mixing and sorting conditions in which response detection capability spanned a 1000-fold range of peptide concentration. With single-cell readout systems for protein expression libraries, this technology offers the promise of screening millions of discrete compound interactions per day. (Journal of Biomolecular Screening 2004:103-111)

Key Words: drug discovery • flow cytometry • automation • cell sorting • sample handling

Journal of Biomolecular Screening, Vol. 9, No. 2, 103-111 (2004)
DOI: 10.1177/1087057103262335
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Home page
 Mol. Pharmacol.Home page
B. S. Edwards, C. Bologa, S. M. Young, K. V. Balakin, E. R. Prossnitz, N. P. Savchuck, L. A. Sklar, and T. I. Oprea
Integration of Virtual Screening with High-Throughput Flow Cytometry to Identify Novel Small Molecule Formylpeptide Receptor Antagonists
Mol. Pharmacol., November 1, 2005; 68(5): 1301 - 1310.
[Abstract] [Full Text] [PDF]

Home page
 J Biomol ScreenHome page
S. M. Young, C. Bologa, E. R. Prossnitz, T. I. Oprea, L. A. Sklar, and B. S. Edwards
High-Throughput Screening with HyperCyt(R) Flow Cytometry to Detect Small Molecule Formylpeptide Receptor Ligands
J Biomol Screen, June 1, 2005; 10(4): 374 - 382.
[Abstract] [PDF]