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Apoptosis detection by annexin V and,,,active caspase-3 with the Agilent 2100,,,bioanalyzer

This Application Note describes how the Agilent 2100 bioanalyzer can be used to analyze apoptotic cell samples. Performance of two assays for apoptosis detection is demonstrated and the data are compared to those obtained with a conventional flow cytometer. Histogram quality of the low number of fluorescent cells counted with the microfluidic sytem is in good agreement with data obtained with the flow cytometer. Detailed protocols and reagent recommendations for an annexin V assay and a caspase-3 assay are given. High reproducibility of chip results, low cell consumption and ease-of-use are advantages the compact Agilent 2100 bioanalyzer system offers for apoptosis cell assays.
The Agilent 2100 bioanalyzer was introduced by Agilent Technologies as the first commercially available lab-on-a-chip analysis system for the life science laboratory using LabChip products, developed by Caliper Technologies Corp. Chip-based approaches for a variety of separation-based techniques have been introduced, addressing DNA, RNA, and protein separations. The Agilent 2100 bioanalyzer is capable of two-color fluorescence detection and runs disposable microfluidic glass chips. The applications presented here are based on the controlled movement of cells by pressure-driven flow inside the interconnected networks of microfluidic channels. Cells are hydrodynamically focused in these channels before passing the fluorescence detector in single file. Each chip accommodates up to six samples and data acquisition of all samples is fully automated while data analysis allows for user-specific settings. Specific advantages of the instrument are the low number of cells required for analysis and the easeof- use. Here, the detection of apoptotic cell death is investigated.

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