Modified Nile red staining method for improved visualization of neutral lipid depositions in stratum corneum.
The neutral lipids existing in the intercellular spaces of the stratum corneum (SC) provide a permeability barrier to prevent water loss. Nile red is the most sensitive lipid stain for tissue sections. However, due to the extremely flattened morphology of corneocytes and the resolution limits of the light microscope, Nile red staining is seldom used as a fluorescent probe for the lipid-rich SC. In this study, we modified the traditional method for visualization of intracellular lipid by adding 4% potassium hydroxide after Nile red staining. This modified method not only allowed visualization of lipids existing in the intercellular membrane regions and the lateral junctions of the adjoining corneocytes, but also clearly demonstrated small lipid droplets within pathological corneocytes. These features were not observed with the traditional staining method. Thus, this modified Nile red staining method greatly improved the resolution of the SC lipids under light microscopy and should be useful for studying lipid depositions in both normal and pathological SC.
Sheu
Hamm-Ming
HM
Department of Dermatology, College of Medicine, National Cheng Kung University Hospital, Tainan, Taiwan.
Tsai
Jui-Chen
JC
Lin
Tzu-Kai
TK
Wong
Tak-Wah
TW
Lee
J Yu-Yun
JY
D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000078404:Epidermal Cells; D004817:Epidermis / Q000737:chemistry; D005260:Female; D005456:Fluorescent Dyes; D006801:Humans; D006878:Hydroxides; D008055:Lipids / Q000032:analysis*; D008297:Male; D008875:Middle Aged; D010078:Oxazines; D017680:Potassium Compounds; D013194:Staining and Labeling / Q000379:methods*
D005456:Fluorescent Dyes; D006878:Hydroxides; D008055:Lipids; D010078:Oxazines; D017680:Potassium Compounds; C044808:nile red; C029943:potassium hydroxide