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. 2014 Mar 10;55(3):1483–1492. doi: 10.1167/iovs.13-13766

Figure 1.

Figure 1

Methods used for quantification. (A) Image showing mGluR6 staining. To compare the average staining intensity in rod bipolar dendritic tips versus that in ON cone bipolar dendritic tips, we drew ROIs around these structures. Puncta in the upper part of the OPL represent the dendritic tips of rod bipolar cells (small rectangles) and characteristic row of puncta lower in the OPL represent the dendritic tips of ON cone bipolar cells (dotted irregular regions). The mean intensity was taken as the average per pixel in the ROIs minus the average background (taken from the ONL, dotted square). These averages were computed from single confocal images. (B) Same image as in (A). Average staining intensity per pixel in the OPL was calculated by encircling the whole OPL (dotted rectangle) and subtracting the average background taken from the ONL (as in [A]). This operation was computed from the captured volume (approximately 4-μm depth). (C) A hypothetical 3D surface showing the intensity (as height) of each pixel in the stained puncta in the x-y dimension (horizontal plane). The dashed black line shows the level of half maximum intensity, and the dark circle indicates the size of the puncta (FWHM). The punctum intensity was calculated as the average of the intensities within the circle above the half maximum width (darker shade).