title: Live endothelial cells imaged by Scanning Near-field Optical Microscopy (SNOM): capabilities and challenges.
abstract: The scanning near-field optical microscopy (SNOM) shows a potential to study details of biological samples, since it provides the optical images of objects with nanometric spatial resolution (50-200 nm) and the topographic information at the same time. The goal of this work is to demonstrate the capabilities of SNOM in transmission configuration to study human endothelial cells and their morphological changes, sometimes very subtle, upon inflammation. Various sample preparations were tested for SNOM measurements and promising results are collected to show: 1) the influence of α tumor necrosis factor (TNF-α) on EA.hy 926 cells (measurements of the fixed cells); 2) high resolution images of various endothelial cell lines, i.e. EA.hy 926 and HLMVEC (investigations of the fixed cells in buffer environment); 3) imaging of live endothelial cells in physiological buffers. The study demonstrate complementarity of the SNOM measurements performed in air and in liquid environments, on fixed as well as on living cells. Furthermore, it is proved that the SNOM is a very useful method for analysis of cellular morphology and topography. Changes in the cell shape and nucleus size, which are the symptoms of inflammatory reaction, were noticed in TNF-α activated EA.hy 926 cells. The cellular structures of submicron size were observed in high resolution optical images of cells from EA.hy 926 and HLMVEC lines.
authors.affiliation: Faculty of Chemistry, Jagiellonian University, Ingardena 3, Krakow, Poland.|Jagiellonian Centre for Experimental Therapeutics (JCET), Bobrzynskiego 14, Kraków, Poland.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Bulat
authors.affiliation: Jagiellonian Centre for Experimental Therapeutics (JCET), Bobrzynskiego 14, Kraków, Poland.
authors.forename: Anna
authors.initials: A
authors.lastname: Rygula
authors.affiliation: Faculty of Chemistry, Jagiellonian University, Ingardena 3, Krakow, Poland.
authors.forename: Ewelina
authors.initials: E
authors.lastname: Szafraniec
authors.affiliation: Kwasei Gakuin University, 2-1 Gakuen, Sanda, Hyougo, 669-1337, Japan.
authors.forename: Yukihiro
authors.initials: Y
authors.lastname: Ozaki
authors.affiliation: Faculty of Chemistry, Jagiellonian University, Ingardena 3, Krakow, Poland.|Jagiellonian Centre for Experimental Therapeutics (JCET), Bobrzynskiego 14, Kraków, Poland.
authors.forename: Malgorzata
authors.initials: M
authors.lastname: Baranska
mesh_terms: D002460:Cell Line; D042783:Endothelial Cells / Q000166:cytology*; D006801:Humans; D008853:Microscopy / Q000379:methods*; D061848:Optical Imaging / Q000379:methods*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: D014409:Tumor Necrosis Factor-alpha
keywords: TNF-α; enhothelium; live cells; scanning near-field optical microscopy (SNOM)