title: Phasor analysis of multiphoton spectral images distinguishes autofluorescence components of in vivo human skin.
abstract: Skin contains many autofluorescent components that can be studied using spectral imaging. We employed a spectral phasor method to analyse two photon excited autofluorescence and second harmonic generation images of in vivo human skin. This method allows segmentation of images based on spectral features. Various structures in the skin could be distinguished, including Stratum Corneum, epidermal cells and dermis. The spectral phasor analysis allowed investigation of their fluorescence composition and identification of signals from NADH, keratin, FAD, melanin, collagen and elastin. Interestingly, two populations of epidermal cells could be distinguished with different melanin content.
authors.affiliation: Utrecht University, Department of Molecular Biophysics, Utrecht, The Netherlands.
authors.forename: Farzad
authors.initials: F
authors.lastname: Fereidouni
authors.forename: Arjen N
authors.initials: AN
authors.lastname: Bader
authors.forename: Anne
authors.initials: A
authors.lastname: Colonna
authors.forename: Hans C
authors.initials: HC
authors.lastname: Gerritsen
mesh_terms: D003094:Collagen / Q000378:metabolism; D020405:Dermis / Q000166:cytology* / Q000378:metabolism*; D004549:Elastin / Q000378:metabolism; D000078404:Epidermal Cells; D004817:Epidermis / Q000378:metabolism*; D005182:Flavin-Adenine Dinucleotide / Q000378:metabolism; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D007633:Keratins / Q000378:metabolism; D008297:Male; D008543:Melanins / Q000378:metabolism; D036641:Microscopy, Fluorescence, Multiphoton; D009243:NAD / Q000378:metabolism
chemical_list: D008543:Melanins; D009243:NAD; D005182:Flavin-Adenine Dinucleotide; D007633:Keratins; D003094:Collagen; D004549:Elastin
keywords: fluorescence microscopy; human skin; image analysis; multispectral and hyper spectral imaging; spectral unmixing; tissue imaging