Troubleshooting ApoTome acquisitions

You may notice residual lines or noise in the image. Follow our instructions to troubleshoot these problems and obtain the sharpest image.

Residual lines

Residual lines in the image often result from a bad calibration. To resolve this issue, we suggest that you try these steps: re-calibrate based on the combination objective/filter cube (see "grid-phase calibration" in ApoTome setup); a mirror calibration is usually not needed.

  1. Verify that the grid lines are moving. If they are not:
    1. Close the software, restart the Apotome power supply box, and then re-open the software.
    2. Re-capture the image.
  2. If the grid is already moving, the SABC settings in the Apotome Setup window: Under Image filter options, check SABC and set to Medium, or increase SABC to Strong if it is already enabled.
  3. If the grid lines are not clearly defined with good contrast between the grids and the tissue, re-calibrate the current objective/filter cube combination (see Grid focus section in ApoTome setup).

Noise

The ApoTome removes out-of-focus light. In cases where the light in the image is mostly out of focus or the starting image is dim, there will be little light left in the ApoTome image. Brightening a dim ApoTome image for display can lead to the image appearing noisy.

To decrease noise, increase the camera's dynamic range to obtain a camera histogram that is as wide as possible, similar to this:

There are two methods to do this:

  • Turn up the light source intensity.
  • Increase the exposure time, gain, or offset (see Camera settings).

Moving the point slider in the histogram makes the image look better on-screen, but has no effect on the actual quality of the ApoTome image.