Figure 6 of Bassnett, Mol Vis 2010; 16:2294-2300.

Figure 6. Quantification of epithelial nuclei in orthographic projections of the equatorial lens region. A: Projection of double-labeled lens tissue showing Edu-positive nuclei (yellow) and Draq5-stained nuclei (red). B: Three-dimensional editing of the original image stack removes the underlying fiber cell nuclei and clarifies epithelial nuclear fluorescence, facilitating automated counting. An arrow indicates the border between the epithelial cell nuclei and those of young fiber cells in the meridional rows (MR). C: Epithelial nuclei are identified automatically and color-coded according to size. Note the densely packed nuclei (blue green color) near the border of the epithelium. D: An isosceles trapezoid is defined in the center of the projection. The width of the trapezoid corresponds to 10° of longitude (See Figure 2). Sides a and b are oriented parallel to the lens equator. Side a is positioned level with the fiduciary cell, d (see Figure 4) and side b is located at the border between the epithelium and the meridional row cells. Nuclei (green) with centroids lying within the trapezoid are included in the count.