Fig. 3. Use of acridine orange to delineate the borders of the organelle-free zone (OFZ) in the developing chicken lens. (A) Midsagittal section of an E16 chicken lens. The lens is shown with its anterior surface facing up. The bulk of the lens is composed of elongated fiber cells arranged in concentric layers. Nuclei (arrow) are present in the peripheral cells but are absent from the oldest, innermost, fiber cells (asterisk). (B) Confocal image of the section shown in A, following incubation in 1 µg ml^–1 acridine orange. Acridine fluorescence is strongest in the peripheral fiber cells. A clearly demarcated region with very low fluorescence is present in the center of the lens (arrowhead). This region corresponds to the OFZ. Scale bar, 250 µm. (C) Tracing of the section shown in B, indicating the spatial parameters that were measured in this study to analyze the time course of organelle breakdown in the developing lens. d, distance; D, diameter; T, thickness.

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