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FIG 4. Immunohistochemical section at a location similar to that in Figure 2 (LCM29 stain for glial keratin; original magnification, x10). Axonal fiber diameters are 1–40 µm. Individual giant reticulospinal axons are located in the lateral region (Mauthner axon) and ventral columns (Müller axons). In the fiber tracts, spaces not occupied by axons are filled with glial cells and their processes. Threshold between negatively stained area (axoplasm) and positively stained area (glial cells and fibers) is selected manually. Image is converted to a binary image, as shown. Average of two threshold determinations is displayed in overlay regions in the areas measured. Percentage of positive staining is calculated in dorsal, lateral, and ventral columns. The Mauthner axon is not included in the lateral columns sampled for MTR measurement.
