Fig. 6

Double immunofluorescence staining results of ChAT and CGRP in facial and sural nerve. a representative image of staining with ChAT (red), CGRP (green), and DAPI (blue). Scale bar is 25 μm. Quantification of b area percentage; c mean gray value; and d axon count of positive ChAT/CGRP staining. Mean gray value cut-points of e CHAT to identify motor nerve fascicles and f CGRP to identify sensory nerve fascicles were determined using ROC curve methodology with sensitivity and specificity. g The most accurate cut-point occurred using CHAT/CRCP: a cutpoint of 0.855 yielded 100% sensitivity and 100% specificity identifying motor and sensory nerve fascicles with an area under the ROC curve of 1.000 (P < 0.001)