Fig. 3.

Immunofluorescence image (A) of a hotspot TUNEL (red, B) and DAPI (blue, C) region prior to cropping. The white box shows the region of analysis used in (B–H). The merged cropped region (D) was processed with the primary four methods: the ImagePro method (E), the RA Toolkit standard setting (F), the RA Toolkit high setting (G), and the MCT method (H). The corresponding individual total cell counts acquired with each method are shown in (I) where ‘E’ refers to ImagePro, ‘F’ refers to the RA Toolkit standard setting, ‘G’ refers to the RA Toolkit high setting, and ‘H’ refers to the MCT method. Scale bar denotes 50μm. Abbreviations: ONL – outer nuclear layer, INL – inner nuclear layer, GCL – ganglion cell layer.