Agilent Seahorse XF Cell Energy Phenotype Test Kit FAQs

What rate measurements are used to calculate assay parameters?

Parameter equations are described in Table 1 on page 5 of the user guide.

How do I remove wells from the Report Generator calculations?

Outlier assay wells must be turned OFF or reassigned in Wave 2.3 before Excel export. See the Exclude Outliers/Groups section from Analysis or the Wave user guide for more information.

How many Excel files can be imported for analysis?

• XF/XFe96 & XF/XFe24: A maximum of five Excel files (*.xlsx) can be imported to the Multi-File XF Report Generator.
• XFp: A maximum of 10 Excel files (*.xlsx) can be imported to the Multi-File XF Report Generator.

Can I import Excel files generated on two different XF analyzers?

No. Assay result data must be generated by the same Agilent Seahorse XF, XFe or XFp Analyzer. Combining data from different analyzers is not supported currently (example: XF96 and XFe96).

Can I use the Excel file exported from the XFp, XF96, or XF24 Analyzer?

Assay result data must be first exported from Wave 2.3 (Desktop or Controller) to an Excel file (*.xlsx), then imported to the Multi-File XF Report Generator. Excel files exported from earlier versions of Wave (Desktop or Controller) and XF96/XF24 software are not compatible. If the Excel file has been exported from Wave but cannot be imported to the Report Generator, please contact Agilent Seahorse Technical Support.

Can I use baseline rate data (%) to calculate assay parameters?

Normalized (or non-normalized)  Oxygen Consumption Rate (OCR) data is used for parameter calculations, parameter calculations using baseline rate data (%) is not supported currently.

If you receive an error message about Instrument Protocol (XFe96; XFe24; XF96; XF24 only)  

Errors upon data import have two likely causes:

• Mismatching Instrument Protocols (see Best Practices)
• A custom cycle in one or more of the imported file Instrument Protocols. A Custom Cycle refers to an extra Mix or Wait command step in the Instrument Protocol for an assay. Custom Cycles are not part of the standardized assay template for the Seahorse XF Cell Energy Phenotype Test and are not supported in Report Generator analysis. Please contact Seahorse Technical Support for any additional questions regarding Custom Cycles.

Where can I find the OCR and ECAR kinetic graphs?

Both the Oxygen Consumption Rate (OCR) and Extracellular Acidification Rate (ECAR) kinetic graph and corresponding data table is found on the Measures Sheet tab in the Seahorse XF Cell Energy Phenotype Report Generator.

More FAQs: Cell Analysis FAQs and Troubleshooting

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