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If you want to install the app on a different system drive follow this support article for additional instruction. Remove the assembled sensor cartridge with Hydro Booster and Utility plate from the incubator. Remove and discard the Hydro Booster. A: image shows low end of optimal cell density, ideal seeding density and distribution.

B: image shows high end of optimal seeding density, some cell clusters can be seen. Place row labels lettered A-H to the left. The triangular notch will be in the bottom left-hand corner. Place row labels lettered A-D to the left. Place well labels lettered A-H to the left. Do not force the tips completely into the holes. Orient the loading guide so the letter 'A' is located in the upper left-hand corner. Use your fingertips to hold the outside edges of the loading guide to stabilize during loading so pipette tips do not dislodge the loading guide.

Position the pipette tips filled with your compounds for injection into the desired column in the loading guide, and orient the tips at a very slight angle. Insert the tips as far as they will go without resistance into the holes and dispense the compound. Using a multichannel pipette, make sure the tips are securely fitted onto the pipette. Place the tips halfway into the injection ports with the bevel of the tip against the opposite wall of the injection port. Do NOT insert the tips completely to the bottom of the injection ports as this may cause compound leakage through the port.

Dispense the compounds into the ports gently. Withdraw the tips from the ports carefully , stabilizing the loading guide throughout the procedure. The liquid should be in the port, make sure there are no residual drops on the top of the sensor cartridge.

View cells under a microscope to: Confirm cell health, morphology, seeding uniformity and purity no contamination. For adherent cells , ensure cells are adhered with a consistent monolayer and were not washed away during washing step.

For suspension cells , ensure cells are stably adhered following centrifugation, washing and incubation. Ensure your background wells do not contain cells. You can verify the instrument connection status in the widget panel in the lower-left corner of Wave Controller software. Double-click to open the desired assay template file from the Templates view. If your assay template is not displayed on the Templates view, transfer the template using a shared network drive or USB flash drive.

After reviewing the group definitions, plate map layout and instrument protocol, click Start Run. After you enter the save location for your result file following completion of the assay , the tray door on the XFe Analyzer will open.

Before starting calibration, ensure: The sensor cartridge fits properly on the Utility plate. The lid is removed from the sensor cartridge. Proper orientation direction of the sensor cartridge on the Utility plate. Place the sensor cartridge hydrated and loaded with compounds and Utility plate onto the tray when prompted. Press I'm Ready to initiate sensor cartridge calibration. Once sensor cartridge calibration is complete, Wave Controller will display the Load Cell Plate dialog.

The sensor cartridge remains inside the XFe Analyzer for this step. Before loading the cell plate, ensure: The lid is removed the Cell Plate.

Proper orientation direction of the Cell Plate on the tray. After completing equilibration, the assay will automatically begin acquiring baseline measurements as outlined in your instrument protocol. Once the final measurement command in the instrument protocol is completed, Wave Controller software will display the Unload Sensor Cartridge dialog.

Click Eject when ready to eject the sensor cartridge and cell plate. Set aside for later analysis if necessary example - cell count normalization. After removing the sensor cartridge and cell plate, the Assay Complete dialog will appear.

Click View Results to immediately open your assay result file, or click Wave Home to return to the Templates view and begin another XFe assay. Save or transfer the assay result file to a shared network drive or USB drive and open using Wave Desktop software on your PC for data analysis.

Procedure to Run your Assay Once all required injection ports are filled, transfer the cartridge and utility plate to the Analyzer and begin cartridge calibration.

When the cartridge calibration is complete, follow the software prompts to exchange the utility plate for the cell culture plate and initiate the XF assay. Touch to open the assay template to review the template design: Group definitions - Touch the group name to display the injection strategy, pretreatments, and cell type information for the selected group.

XFp Analyzer software does not allow modifications to group definitions, Wave Desktop software must be used. Plate Map - Change a well assigned to a group, touch the group name first, then touch the well on the plate map. Touch a template from the list to open and review the group definitions and plate map layout: Group Definitions — Touch the group name to display the injection strategy, pretreatments, assay media, and cell type for the selected group.

Modifications to group definitions can be made using the modify function in Agilent Seahorse Analytics. Plate Map — To change the group assigned to a well on the plate map, first touch the group name from the list then touch the well on the plate map. Touch the right arrow lower-right corner to review or edit the instrument protocol.

Press Edit next to Assay Name to customize the name of your assay result file. Press Edit next to Notes to add custom notes related to your assay. Press Edit next to Email Notification to notify recipients for user-interaction example — replace utility plate with the cell plate , and to automatically send the assay result file following completion of the assay. This feature requires an active internet connection on the XFp Analyzer.

Add notes about the assay. Review alert notifications. Enter an email address to receive automated notifications from the XF HS Mini Analyzer when user interaction is required, and a copy of the assay result file when the assay has completed.

An active internet connection is required for this feature. When ready to begin the XF assay, click Start Assay. Before loading the cell plate, ensure: The sensor cartridge fits properly on the Utility plate. Touch Start Run when you are ready to begin your XF assay. Follow the on-screen prompts to load the calibrant utility plate and sensor cartridge hydrated and loaded with compounds for injection. Before starting calibration, verify: The sensor cartridge fits properly on the calibrant utility plate.

The sensor cartridge lid has been removed. Once calibration is complete the XFp Analyzer tray will open and present the utility plate. Remove the utility plate and load the cell plate on the tray. Once calibration is complete, press Open Tray to present the calibration utility plate. Remove the calibration utility plate and place the cell plate on the tray. Before loading the cell plate to start your assay, verify: The cell plate lid has been removed. After placing the Cell Plate on the tray, touch Continue to begin equilibration.

The first step in your assay is called Equilibration. Once Equilibration is complete, the analyzer will start acquiring the first baseline measurement as defined in your instrument protocol. Above the data display in the center of the view you can see the current and completed commands in your instrument protocol.

Once the final measurement command in the instrument protocol is completed, the Remove Plate and Cartridge dialog will appear. Click Continue to eject the sensor cartridge and cell plate. Once the final measurement in your instrument protocol has completed, press Eject to eject the sensor cartridge and cell plate from the XF HS Mini.

When the tray is fully ejected, remove the sensor cartridge and cell plate from the tray and set aside for additional analysis if necessary example - cell count normalization. Press Continue to close the tray. Select a topic below to learn more: 5. JPG and. Below the files list, you will see My Analyzers, where you can assign one or more Seahorse XF Analyzers to your profile.

To import a data file from the Home view, click the small File Upload button in the upper-right corner above the files list.

Settings and User Information: Click the small user account icon in the upper-right corner of the dark blue top navigation bar to manage your user account information, or logout. Click the Settings and User Data link to display account management options, which include: Checking the amount of free space to store data files, view the Agilent Privacy Policy, or delete your Seahorse Analytics account.

Renaming or deleting your custom analysis views. Changing your Seahorse Analytics account password. Notifications: Click the small bell icon in the upper-right corner of the dark blue top navigation bar to display your account notifications.

Help: Click the question mark icon in the upper-right corner of the dark blue top navigation bar to display basic help information. How to import data files There are two ways to get data files into your account: 1 manually import data files or 2 accepting data files that have been shared with you. Click the File Upload button to display the file upload dialog pictured on the right. Import one or multiple data files at a time, you can see that 2 data files have been selected for import.

Choose a custom folder to save the data file in rather than the main files list. You can always move a data file to a custom folder after you have imported it. Assign a category label to a data file by typing a new category, or by clicking in the category field to display existing category labels.

Click Import Seahorse Analytics will confirm a successful file import. When you dismiss this dialog, you will see the imported file s are displayed first in the files list. Click this 3-dot button to display a menu of additional file features, which include: Send To: Display the share dialog to enter an email address to send the selected file to.

Delete: Delete the selected file from your account. Categories: Add or delete category labels. Make a Copy: Create a copy of the selected file.

Rename: Rename the selected file. Move to folder: Move the selected file to a custom folder. Download: Download the selected file to your PC. Blank and Custom Analysis Views The custom analysis view feature provides ultimate data analysis flexibility. Important: For XF Cell Mito Stress Test assays with 4 injections, you must identify the oligomycin injection using the drop-down menu seen above the widgets before you can add this analysis view.

Select the desired parameter widgets e. A blue outline around the widget indicates it is selected. If the Acute Response widget is greyed-out, then it cannot be selected as this parameter can only be calculated for XF Cell Mito Stress Test assays with an acute injection.

Spare Respiratory Capacity to an existing analysis view: a. From the analysis view, click Modify upper-right corner of the dark blue ribbon at the top of the application. Click the Assay Media tab to confirm buffer factor is assigned to your assay media group definitions. If you are using a custom assay media, you will need to input the buffer factor of your custom media.

Click the Background Buffer tab to confirm buffer factor is assigned to your background wells. Click the Groups tab to confirm the assay medium is assigned to each assay group. Once finished click Save. Once you have performed or corrected the above steps you should be able to add your analysis view or widget to the data file.

Use the Measurement drop-down menu to display group data for another rate measurement in the assay. Export all rate data You can create an Excel and Prism file containing all rate data for the selected file. How to export all rate data: Go to the Files or Home view. Click the 3-dot button seen on the right of the file column to display the menu. Click Export to Excel or Export to Prism to create the desired file. Use the Export to Excel and Export to Prism options found under the menu for individual files on the Home and Files views green highlight, pictured right.

Related Support Material Wave Desktop 2. Select a topic below to learn more: 6. Online store Shop the online store to purchase cartridges, plates, kits, media, and reagents to your your XF assay.

Buy XF Assays. Request a demo. Worldwide technical support E-mail: cellanalysis. Seed cells at single or different densities and visually assess degree of cell confluence; choose a miniplate for the next step.

See method below for preparation. Seahorse Data File Compatibilty. XFe96 assay result file. XFe24 assay result file. XFp assay result file. XF HS Mini assay result file. Internet Browser Compatibility. Microsoft Edge Use of Internet Explorer is strongly discouraged. Microsoft Excel.

Graphpad Prism. Image File. Last rate measurement before first injection — Non-Mitochondrial Oxygen Consumption. Last rate measurement before Oligomycin injection — Minimum rate measurement after Oligomycin injection. Last rate measurement before Oligomycin injection — Last rate measurement before acute injection. Last OCR rate measurement before first injection.