This Application Note describes a simplified and fast protocol for the detection of apoptotic cells with the Agilent 2100 bioanalyzer and the cell fluorescence LabChip®. The specific advantage of the simplified protocol is the amount of time saved and its ease-of-use. The note describes how the Agilent 2100 bioanalyzer can be used to analyze apoptotic cell samples. Annexin V staining of live cells is demonstrated and the data are compared to that obtained with a conventional flow cytometer. Histogram quality of the microfluidic system is comparable to that of a conventional flow cytometer. Detailed protocols and reagent recommendations for staining of cells are provided. The high reproducibility of the chip results, low cell and reagent consumption, the speed and the ease of use are advantages the Agilent 2100 bioanalyzer offers for monitoring apoptotic cells by annexinV staining.
This document presents an optimized, rapid method for analyzing cell apoptosis using the Agilent 2100 bioanalyzer system. The protocol utilizes the Cell Fluorescence LabChip kit to detect apoptotic cells stained with Annexin V and Calcein AM.
The note details a specific workflow involving Jurkat cells treated with Camptothecin or Anti-Fas antibody. It provides step-by-step instructions for preparing reagents, staining samples, and running the analysis using the predefined 'Apoptosis — fast protocol' in the software. Results obtained via this method are shown to be comparable to conventional flow cytometry.
After mixing with Solution A, the cells must be incubated for 10 minutes at room temperature.
The cells must be centrifuged at 400 x g for 2 minutes.