2.5. Histological studies
12cm2 of dorsal skin was excised between fore and hind
legs after topical application with Polygonum multiflorum extract
(4.7 mg/12cm2) at the indicated time points. Dorsal skin was
maintained in 10% neutral formalin and embedded in paraffin
blocks to obtain longitudinal and transverse section. 5m sections
were stained with hematoxylin and eosin (H&E). Digital
photomicrographs were taken from representative areas at a fixed
magnification of 100×.
2.6. Hair follicle count
The H&E stained slides were photographed using a digital photomicrograph
and all of the images were cropped in a fixed area of
300 pixels width. We counted hair follicles manually in a fixed area
(0.09mm2). Digital photomicrographs were taken from representative
areas at a fixed magnification of 100×.
2.7. Hair length determination
Hairs were plucked randomly from shaved dorsal area at 0, 1, 2,
3 and 4 week. After plucking 10 hairs per mouse, we measured the
average hair length manually.