Agilent Seahorse XF Imaging and Normalization System
Agilent and BioTek have joined forces to create the Agilent Seahorse XF Imaging and Normalization System. Seahorse XF Analyzers are now integrated with BioTek's Cytation Cell Imaging Multi-Mode Readers providing an easy to use cell count based normalization solution for XF data comparison. Rapid cell count based normalization enables reliable, validated, meaningful XF data comparison, well to well, plate to plate, and experiment to experiment.
Agilent Seahorse XF technology enables real time measurement of the mitochondrial respiration and glycolysis in live cells. Because we are dealing with the real time activity, determining cell culture status and getting accurate cell counts on plates is critical for correct data interpretation. There are many factors that can lead to a significant well to well cell count variations. It may be due to differences in cell plating efficiency or in sensitivity to contact inhibition.
Also when cells are stimulated biologically or chemically there's a good chance they're proliferation rate or death rate will change. Macrophage activation is a good example in which cell counts can change significantly. In vitro macrophage activation is commonly induced by treatment with lipopolysaccharide alone or together with interferon gamma.
The activation associated metabolic phenotype changes are measured from several hours to overnight. When the samples are ready to be analyzed, the bar code scans of the cell plate registers the plate. Then the plate is placed in the Cytation. The plate can stay in the Cytation to de-gas.
It's a simple four step process. First brightfield scans in the Cytation document sample quality.
Next the plate is placed in the XF Analyzer and the metabolic analysis assay is run. The plate is then placed back in the Cytation where a fluorescence scan counts cells then passes the scan and cell count information to Wave software for normalization. Finally XF metabolic analysis data, normalized cell counts, and brightfield and fluorescence images are linked together in an XF result file ready for analysis and interpretation in Wave.
In the case of the glycolytic rate assay, the data interpretation dramatically changed after the data was normalized.
The Agilent XF Imaging and Normalization System provides a seamless simple workflow integrating the real time imaging capability of Cytation with the XF analysis system.
It automates and accelerates access to data normalization and interpretation resulting in the reliable accurate and meaningful XF data comparison.