Cell Metabolism: Seahorse XF Imaging and Normalization

Agilent offers an integrated imaging and normalization system with the Seahorse XF analyzer. The system combines cellular analysis and imaging for an intuitive workflow to streamline your XF data interpretation. This imaging and normalization solution combine Agilent strength in cellular metabolic analysis with the Agilent BioTek capability in cell imaging.

Figure 1. Cell imaging workflow.

Example of XF data normalization using in situ nuclear staining and in situ cell counting.

Figure 2. SKOV3 cells were plated at 1 x 10⁴, 2 x 10⁴, 3 x 10⁴ cells per well, cultured 24 h, subject to the XF Cell Energy Phenotype Test and followed by image analysis. A) Raw OCR and ECAR change with injection (arrows) of oligomycin + FCCP (1.0 and 0.5 µM final, respectively), including 20 µM Hoechst 33342 (2 µM final). B) Representative images of nuclei fluorescently labeled by Hoechst 33342 (upper panel) and nuclei identified and outlined using the Seahorse XF Imaging and Cell Counting software with the Agilent BioTek Cytation 1 (lower panel). C) OCR and ECAR normalized by in situ nuclear staining cell counts (Mean ± SD, n=4).

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Application Notes:

• XF Data Normalization by the Agilent Seahorse XF Imaging and Normalization System
• Normalization of Agilent Seahorse XF Data by In-situ Cell Counting Using a BioTek Cytation 5

Technical Notes:

• Agilent Seahorse XF Imaging & Cell Counting Software

Related Products:

• Agilent BioTek Cytation 5 Cell Imaging Multi-Mode Reader
• Agilent BioTek Cytation 1 Cell Imaging Multi-Mode Reader
• Agilent Seahorse XF

For Research Use Only. Not for use in diagnostic procedures.

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