title: Label-free and non-invasive discrimination of HaCaT and melanoma cells in a co-culture model by hyperspectral confocal reflectance microscopy.
abstract: A novel hyperspectral confocal microscopy method to separate different cell populations in a co-culture model is presented here. The described methodological and instrumental approach allows discrimination of different cell types using a non-invasive, label free method with good accuracy with a single cell resolution. In particular, melanoma cells are discriminated from HaCaT cells by hyperspectral confocal imaging, principal component analysis and optical frequencies signing, as confirmed by fluorescence labelling cross check. The identification seems to be quite robust to be insensitive to the cellular shape within the studied samples, enabling to separate cells according to their cytotype down to a single cell sensitivity. Set of hyperspectral images of melanoma-keratinocytes co-culture model (left), score plot of principal component analysis and spectral analysis of principal components coefficients (center), label-free spectral identification of cell populations (right).
authors.affiliation: CNR-ISC Istituto dei Sistemi complessi Via fosso del Cavaliere, 100 00133, Rome, Italy.
authors.forename: Francesca R
authors.initials: FR
authors.lastname: Bertani
authors.affiliation: Dermatology Unit, NESMOS Department, Sapienza University of Rome, via di Grottarossa 1035, 00189, Rome, Italy.
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Botti
authors.affiliation: CNR-ISC Istituto dei Sistemi complessi Via fosso del Cavaliere, 100 00133, Rome, Italy.
authors.forename: Luisa
authors.initials: L
authors.lastname: Ferrari
authors.affiliation: CNR-ISC Istituto dei Sistemi complessi Via fosso del Cavaliere, 100 00133, Rome, Italy.
authors.forename: Valentina
authors.initials: V
authors.lastname: Mussi
authors.affiliation: Dermatology Unit, NESMOS Department, Sapienza University of Rome, via di Grottarossa 1035, 00189, Rome, Italy.
authors.forename: Antonio
authors.initials: A
authors.lastname: Costanzo
authors.affiliation: CNR-ISC Istituto dei Sistemi complessi Via fosso del Cavaliere, 100 00133, Rome, Italy.
authors.forename: Marco
authors.initials: M
authors.lastname: D'Alessandro
authors.affiliation: CNR-ISC Istituto dei Sistemi complessi Via fosso del Cavaliere, 100 00133, Rome, Italy.
authors.forename: Francesco
authors.initials: F
authors.lastname: Cilloco
authors.affiliation: CNR-ISC Istituto dei Sistemi complessi Via fosso del Cavaliere, 100 00133, Rome, Italy. stefano.selci@isc.cnr.it.
authors.forename: Stefano
authors.initials: S
authors.lastname: Selci
mesh_terms: D002460:Cell Line; D018920:Coculture Techniques; D006801:Humans; D015603:Keratinocytes / Q000166:cytology*; D008545:Melanoma / Q000473:pathology*; D018613:Microscopy, Confocal; D025341:Principal Component Analysis
keywords: hyperspectral microscopy; melanoma cells HaCaT cells co-culture; multivariate analysis