SPA III Quick Reference Guide for Custom Panels

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1 SPA III Quick Reference Guide for Custom Panels This guide contains instructions for using the BD FACS Sample Prep Assistant III (SPA III) with BD FACS SPA software version.0 and later. Initial Setup Custom panels can be created in any user account with administrative privileges in SPA software. Once created, the custom panels can be used by all user accounts. If you plan to import SPA worklists into BD FACSDiva software, you must create identical panels in both SPA software and BD FACSDiva software. Creating a Custom Panel Start SPA software and log in. Select Setup > Panel Editor. Create a new category if needed. Create a new panel and rename it. Select Other Panel. Creating Tubes for a Custom Panel With the custom panel selected, click New Tube. Edit the new tube that appears. Specify a tube name, and sample and lyse volumes. Access the reagent list and drag reagents, one at a time, from the Reagent List to the Reagent field in the Tube Editor. Click Reagent List to access the Reagent List. Add reagents to the list as needed Rev. A

2 Specify an appropriate volume for each reagent in the Tube Editor. Select a reagent, then specify a volume. Repeat for each reagent. 9 0 In the Tube Editor, click Apply. Click Close to close both the Tube Editor and the Reagent List. Repeat steps through to create additional tubes for the custom panel. In the Panel Editor, select the custom panel and specify the reagent and lyse incubation times. Click OK to close the Panel Editor. Creating a Custom Reagent Rack Select Setup > Reagent Rack Editor. Create a new rack and rename it. With the new rack selected, click Edit Rack. Click Reagent List to display the Reagent List. Drag reagents, one at a time, from the Reagent List to the Rack Map Editor. Specify the rack position for each reagent. Select a reagent, then click a position on the rack map to assign it. Repeat until all reagents have been assigned to a position on the map. Add a rack and specify the position of reagents on the rack, if needed. Close the Rack Map Editor and the Reagent Rack Editor Rev. A

3 Using a Custom Panel The following figure shows the daily workflow steps for using a custom panel. Start Up System Set Up Worklist Run Worklist Shut Down System Starting Up the System Check for leaks or mineral deposits around the syringes, valves, and pumps that are located on the back of the instrument. 9 0 Make sure there are no crimps in the fluidics tubing. Fill the tanks with DI water and lysing solution, if necessary. Empty the waste tank and add L of undiluted bleach and 00 µl of Sigma Antifoam A Concentrate to the tank. Lock the primary tube rack cage. Close the safety cover. Turn on the instrument power. Turn on the computer. Start the software and log in. After the initialization process is complete, check for bubbles and manually prime the fluid lines, if needed Rev. A

4 Setting Up a Worklist Select File > New Prep Worklist to open a blank worklist. Enter sample information. A sample ID is required. Ensure that the appropriate tube type is selected to prevent damage to the instrument. Select the appropriate custom panel. Make Worktable assignments. Select the appropriate reagent rack ID. Select to skip the initial sample mix or to incubate offline, as needed. Select the appropriate carousel ID. Enter a unique carousel ID if needed. Enter lot information if needed. Running a Worklist Load the primary and secondary tubes into the instrument. Place the reagent vials in the reagent rack and load the rack into the instrument. Uncap the reagent vials and store the caps in their corresponding positions in the numbered reagent cap rack. Close the safety cover. Click Run in the software. Complete the verifications stated in the dialog that appears and then click OK. Click OK again to save the worklist. Choose to print the worklist or click Close Rev. A

5 Shutting Down the System Open the safety cover. Remove the carousel. Uncap the vial in R of the reagent rack and fill it with BD FACSClean solution or 0% bleach. Place the vial back in R. Close and lock the primary tube rack. Close the safety cover. From the main menu in the Prep Worklist window, select File > Shutdown. Select the Daily Clean checkbox, then click OK. 9 Click Yes. After the cleaning is complete, click OK to close the software. 0 Turn off the instrument and computer Rev. A

6 Transferring the Worklist to a BD FACSCanto II Workstation Follow this procedure to copy a saved worklist to an external storage device and then import the worklist into BD FACSDiva software. See the BD FACS Loader User s Guide for details about importing the worklist into BD Worklist Manager software. Importing a Worklist into BD FACSDiva Software Copy the saved worklist from the SPA workstation to a portable USB device. Plug the USB device into the BD FACSCanto II workstation. Start BD FACSDiva software and log in. Select File > Import > Worklist. Navigate to the USB device and select the worklist to import. Click Import. Verify that the imported information is accurate Rev. A