Table 3.

Best practices for quantifying neurogenesis

Use appropriate high numerical aperture oil immersion objective lenses for the best axial resolution (i.e., the narrowest depth of field).

Validate staining exists throughout the mounted section thickness so that there is no “dead zone” in the middle devoid of staining where no cells will be counted, although they may exist there.

Use design-based stereological sampling to estimate total cell number independent from differences in region or volume between experimental and control conditions.

Determine the thickness of mounted (not cut) tissue for use in calculating results.

Use careful systematic sampling of the tissue when collecting image stacks to ensure compliance with stereological formulas.

Confocal image stacks are ready-made for stereological counting, but use guard zones and dissector probe counting rules to avoid artifacts and methodological biases.