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Acquire Image Stack

An image stack is a series of single images placed on top of each other. Acquiring image stacks instead of single images adds a z-dimension to the images, and is critical for certain probes3D neuron reconstruction.

1. Click Acquisition>Acquire Image Stack to display the Image Acquisition window.
2. To use different acquisition parameters, click the Options button to open the Image Stack Acquisition Options window.
   • Inputs to define height and location: Select Use Z position at time of acquire ONLY if you've already focused to the top of the stack to be captured (if you are not focused at the top, the objective could hit the slide).
   • Distance between images: Must be greater than the z-resolving power of the microscope (i.e., 0.2µ).
   • Delay before each image: Refers to the time allowed for the stage to settle after a Z move. If there is evidence of motion in the images, try 200ms.
3.  After selecting different parameters, the Image Acquisition window changes to reflect the new parameters. For example, .
   • To use the Set button, place a reference point first.
4. Click the Live Image button to easily obtain a live feed from your camera (same as Acquisition>Live Image).
5. Click the Acquire Single Image button to test your settings (e.g., exposure) on a single image before collecting the entire stack.
   • This button is the equivalent of Acquisition>Acquire Image.
   • If you selected the Multichannel acquire box, it is the same as using Acquisition>Acquire Multichannel Image.

   To use Multichannel acquire, your system must be set up correctly (see Automating the Multichannel Acquisition Process)

See Image Stack overview

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