Real-time fluorescence detection of multiple microscale cell culture analog devices in situ

✍ Scribed by Taek-il Oh; Jong Hwan Sung; Daniel A. Tatosian; Michael L. Shuler; Donghyun Kim

• Publisher: John Wiley and Sons
• Year: 2007
• Tongue: English
• Weight: 483 KB
• Volume: 71A
• Category: Article
• ISSN: 0196-4763
• DOI: 10.1002/cyto.a.20427

✦ Synopsis

Abstract

Background:

We investigated multiple microscale cell culture analog (μCCA) assays in situ with a high‐throughput imaging system that provides quantitative, nondestructive, and real‐time data on cell viability. Since samples do not move between measurements, captured images allow accurate time‐course measurements of cell population response and tracking the fate of each cell type on a quantitative basis. The optical system was evaluated by measuring the short‐term response to ethanol exposure and long‐term growth of drug‐resistant tumor cell lines with simultaneous samples.

Methods:

The optical system based on epi‐fluorescent excitation consists of an LED and a CCD as well as discrete optical components for imaging a large number of cells simultaneously. HepG2/C3A and MESSA cell lines were cultured in two μCCA systems for continuous cell status monitoring in cell death experiments with ethanol and long‐term cell growth.

Results:

The experiment that tested ethanol uptake showed that ethanol immediately caused cell death. The system was applied to extracting dynamic constants in the uptake process. In the long‐term cell growth experiment, growth of MESSA cells was followed by a stationary phase and eventual cell death attributed to nutrient and oxygen depletion and a change in the pH because of the accumulation of wastes by cell metabolism. HepG2/C3A cells were subject to contact inhibition and cell number did not change significantly over time. Issues related to long‐term assays are also discussed.

Conclusions:

The quantitative results have been consistent with qualitative images and confirm the applicability of the portable optical system, and potential application to high‐throughput analysis of cell‐based assays to measure long‐term dynamics. © 2007 International Society for Analytical Cytology