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In this how-to video, we will cover how to set up an image acquisition using the z-stacking feature in CellReporterXpress First set up focus as normal in the acquisition setting step and snap a preview of your image to best visualize the single plane image toggle on and off the available channels in the image viewer. This also aids in optimizing the settings for exposure and focus offset to view the z-stacking option, click on the z-stacking icon by default. The z-stacking option is turned off in the bottom pane, set z-stacking to on the recommended focus step - the distance between planes and the z-stack - is automatically generated for each objective and is determined by the depth of field of the objective. If desired, the focus step value can be changed manually. The Z steps and the z-stack will be centered around the focus offsets configured in the first acquisition set up step. This can be seen by the highlighted focus plane in the middle of the z-stack preview bar located in t