Real-time quantitative fluorescence measurement of microscale cell culture analog systems

Taek Il Oh, Donghyun Kim, Daniel Tatosian, Jong Hwan Sung, Michael Shuler

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

A microscale cell culture analog (μCCA) is a cell-based lab-on-a-chip assay that, as an animal surrogate, is applied to pharmacological studies for toxicology tests. A μCCA typically comprises multiple chambers and microfluidics that connect the chambers, which represent animal organs and blood flow to mimic animal metabolism more realistically. A μCCA is expected to provide a tool for high-throughput drug discovery. Previously, a portable fluorescence detection system was investigated for a single μCCA device in real-time. In this study, we present a fluorescence-based imaging system that provides quantitative real-time data of the metabolic interactions in μCCAs with an emphasis on measuring multiple μCCA samples simultaneously for high-throughput screening. The detection system is based on discrete optics components, with a high-power LED and a charge-coupled device (CCD) camera as a light source and a detector, for monitoring cellular status on the chambers of each μCCA sample. Multiple samples are characterized mechanically on a motorized linear stage, which is fully-automated. Each μCCA sample has four chambers, where cell lines MES-SA/DX-5, and MES-SA (tumor cells of human uterus) have been cultured. All cell-lines have been transfected to express the fusion protein H2B-GFP, which is a human histone protein fused at the amino terminus to EGFP. As a model cytotoxic drug, 10 μM doxorubicin (DOX) was used. Real-time quantitative data of the intensity loss of enhanced green fluorescent protein (EGFP) during cell death of target cells have been collected over several minutes to 40 hours. Design issues and improvements are also discussed.

Original languageEnglish
Title of host publicationImaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues V
Volume6441
DOIs
Publication statusPublished - 2007 Apr 30
EventImaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues V - San Jose, CA, United States
Duration: 2007 Jan 222007 Jan 24

Other

OtherImaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues V
CountryUnited States
CitySan Jose, CA
Period07/1/2207/1/24

All Science Journal Classification (ASJC) codes

  • Electronic, Optical and Magnetic Materials
  • Biomaterials
  • Atomic and Molecular Physics, and Optics
  • Radiology Nuclear Medicine and imaging

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  • Cite this

    Oh, T. I., Kim, D., Tatosian, D., Sung, J. H., & Shuler, M. (2007). Real-time quantitative fluorescence measurement of microscale cell culture analog systems. In Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues V (Vol. 6441). [64410R] https://doi.org/10.1117/12.700610