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3.7. Why do my objects appear elongated in the Z axis

Commonly when viewing your fluorescently labelled structures, aberration in Z is caused by a mismatch between the physical and optical step between each Z layer.

In the first instance it is important to ensure that the spatial calibration of the objective lenses has been performed correctly. This is covered on pages 120 and 121 of the Volocity userguide.

Elongation in Z is a common problem in Optical Microscopy, the factors affecting axial elongation are discussed both in the Volocity Userguide (page 355-356) and in Technical Note 382.

Additional References:

Kam, Z. et al., 2001, Computational adaptive optics for live threedimensional biological imaging, PNAS, v. 98, p. 3790-3795

Scalettar, B. A. et al, 1996, Dispersion, aberration and deconvolution in multi-wavelength fluorescence images, Journal of Microscopy, v. 182, p. 50-60

Sherman, J. et al, 2002, Adaptive correction of depth-induced aberrations in multiphoton scanning microscopy using a deformable mirror, Journal of Microscopy, v. 206, p. 65-71