Why Do My Objects Appear Elongated in the Z Axis?

Why Do My Objects Appear Elongated in the Z Axis?

Why Do My Objects Appear Elongated in the Z Axis?

Elongation in the Z axis is a common issue in optical microscopy and is often caused by a mismatch between the physical and optical step size between Z layers during imaging.

Steps to Address Z-Axis Elongation:

  1. Verify Spatial Calibration:

    • Ensure that the spatial calibration of the objective lenses has been performed correctly.
    • Detailed instructions are available in the Volocity User Guide (pages 120-121).

  2. Understand Optical Aberrations:

    • Elongation in Z can result from axial aberrations inherent to optical microscopy.
    • These factors are discussed in the Volocity User Guide (pages 355-356) and in Technical Note 382.

Additional References on Z-Axis Aberrations:

  • Kam, Z. et al. (2001). Computational adaptive optics for live three-dimensional biological imaging. PNAS, 98, 3790-3795.
  • Scalettar, B. A. et al. (1996). Dispersion, aberration, and deconvolution in multi-wavelength fluorescence images. Journal of Microscopy, 182, 50-60.
  • Sherman, J. et al. (2002). Adaptive correction of depth-induced aberrations in multiphoton scanning microscopy using a deformable mirror. Journal of Microscopy, 206, 65-71.

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