TABLE 1:

Blinded ratings of digitally stained confocal micrographs

Evaluation category	Assessment	Rating	AO	DRAQ5	p-value
Perceived Image Quality	Preferred digital H&E staining	DRAQ5 or AO	18% (10–25%)	82% (75–90%)	<.0001
	Nuclear enhancement	1–10 (poor-excellent)	4 (2.5–5)	7 (5.5–8)	<.0001
	Brightness, contrast, and saturation	1–10 (poor-excellent)	5 (4–7)	7 (5.5–7.5)	<.0001
	Background clarity	1–10 (poor-excellent)	4 (4–5)	6 (5–8)	<.0001
	Staining homogeneity	1–10 (poor-excellent)	4 (3–5)	6 (5–7)	<.0001
Histopathological Features	Preferred feature visualization	DRAQ5 or AO	19% (11–27%)	81% (73–89%)	<.0001
	Epidermis	Presence	74% (65–83%)	85% (78–93%)	<.0001
	Dermal fibers	Presence	75% (66–84%)	72% (63–81%)	<.0001
	Adipocytes	Presence	31% (22–40%)	31% (22–41%)	0.0002
	Hair follicles	Presence	78% (70–87%)	83% (76–91%)	<.0001
m	Hair shafts	Presence	39% (29–49%)	30% (21–40%)	0.0033
	Sebaceous glands	Presence	59% (49–69%)	64% (54–73%)	0.0218
	Sweat glands	Presence	53% (42–63%)	53% (42–63)	0.6080
	Tumors	Presence	19% (11–27%)	17% (9–25%)	<.0001
	Inflammatory cells	Presence	60% (50–70%)	82% (75–90%)	<.0001
	Blood vessels	Presence	18% (9–25%)	44% (34–54%)	<.0001

Chart for paired, blinded evaluation of digitally stained images for perceived image quality and histopathological feature identification. Values presented as percentages (lower and upper confidence limits) or medians (interquartile range).

H&E: Hematoxylin and Eosin; DRAQ5: Deep red anthraquinone 5; AO: Acridine orange