Settings to inject enough water with the micro-pump (SP8-FLIM) under the 25x IMM objective at 37c. These settings have been tested for 8 hours. # Switch ON the pump (button at the back) # Start the software "AOI Controller" - Tab "Settings", select "Serial Port (USB)" = COM8 (or check what's written on the green tape on top of the controller) - Tab "Settings" select "Pump amplitude for interval water flow" = 75 volts - Tab "Experiment": - Check "Activate water flow (pump interval) every" - "Active water flow pump interval, every" = 20 minutes. - "Water time at each time port" = 9 sec. - You can start the pump whenever the experiment is ready with the button "Start Pump" # On the controller the values should be: - HRZ = 9 - Vpp = 95 - In SRS mode - You can change these values with the switches on the controller # Setup your sample with the 25x IMM objectif - Check that there is distilled water in the water reservoir - Initiate the flow with the pump until you see water flowing out of the objective - Add 300 uL of distilled water on the objective - Add your sample (Coverslip) on the water - Find your focus - You can start the pump whenever the experiment is ready with the button "Start Pump" - Check again the settings displayed on the water pump controller NB : unfortunately the software cannot be switched off normally, just use Alt+Ctrl+Del and terminate the process named AOIControler.exe

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