Acquiring an image stack

Requires a separate license

Purpose

An image stack is a series of single images, representing z-planes of the specimen, stacked on top of each other. Acquiring image stacks adds a z-dimension to the images, and is critical for 3D neuron reconstruction.

Procedure

Go to the Acquire ribbon and click Stack acquisition.
To use different acquisition parameters, click the Options button.
- 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., 0.2 µ).
- 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 200 ms.
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.
Click Acquire>Display>Live Image to show a live feed from your camera onscreen.
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 Acquire>Capture>Acquire Image.
- If you selected the Multichannel acquire box, it is the same as using Acquire>Capture>Enable multichannel.