Real-Time Assessment of the Antifungal Activity of Primary Human Immune Cells

To visualize DsRed and AlexaFlour 633 during microscopy, preheat a microscope heater and turn on the microscope and the computer. Next, add 100 microliters of 3 times 10 to the 6th per milliliter FLARE conidial suspension to the appropriate wells of the imaging dish. Record the time that FLARE conidia are added.

Mount the imaging dish on the microscope stage. Then, in the Acquisition settings, set the Laser Power to 10% and Exposure Time to one second, and adjust the camera sensitivity for a clear but not overexposed image of DsRed and AF 633, and set up a list of stage points. Initialize imaging when all points are in focus, the fluorescent channels are optimized, and the cycle time and duration are set.