title: Morphology and force probing of primary murine liver sinusoidal endothelial cells.
abstract: Liver sinusoidal endothelial cells (LSECs) represent unique type of endothelial cells featured by their characteristic morphology, ie, lack of a basement membrane and presence of fenestrations-transmembrane pores acting as a dynamic filter between the vascular space and the liver parenchyma. Delicate structure of LSECs membrane combined with a submicron size of fenestrations hinders their visualization in live cells. In this work, we apply atomic force microscopy contact mode to characterize fenestrations in LSECs. We reveal the structure of fenestrations in live LSECs. Moreover, we show that the high-resolution imaging of fenestrations is possible for the glutaraldehyde-fixed LSECs. Finally, thorough information about the morphology of LSECs including great contrast in visualization of sieve plates and fenestrations is provided using Force Modulation mode. We show also the ability to precisely localize the cell nuclei in fixed LSECs. It can be helpful for more precise description of nanomechanical properties of cell nuclei using atomic force microscopy. Presented methodology combining high-quality imaging of fixed cells with an additional nanomechanical information of both live and fixed LSECs provides a unique approach to study LSECs morphology and nanomechanics that could foster understanding of the role of LSECs in maintaining liver homeostasis.
authors.affiliation: Centre for Nanometer-Scale Science and Advanced Materials (NANOSAM) Faculty of Physics, Astronomy and Applied Computer Science, Jagiellonian University, Cracow, Poland.
authors.forename: B
authors.identifier: 0000-0003-2129-3714
authors.initials: B
authors.lastname: Zapotoczny
authors.affiliation: Centre for Nanometer-Scale Science and Advanced Materials (NANOSAM) Faculty of Physics, Astronomy and Applied Computer Science, Jagiellonian University, Cracow, Poland.
authors.forename: K
authors.initials: K
authors.lastname: Owczarczyk
authors.affiliation: Centre for Nanometer-Scale Science and Advanced Materials (NANOSAM) Faculty of Physics, Astronomy and Applied Computer Science, Jagiellonian University, Cracow, Poland.|Jagiellonian Centre for Experimental Therapeutics (JCET), Jagiellonian University, Cracow, Poland.
authors.forename: K
authors.initials: K
authors.lastname: Szafranska
authors.affiliation: Jagiellonian Centre for Experimental Therapeutics (JCET), Jagiellonian University, Cracow, Poland.
authors.forename: E
authors.initials: E
authors.lastname: Kus
authors.affiliation: Jagiellonian Centre for Experimental Therapeutics (JCET), Jagiellonian University, Cracow, Poland.|Chair of Pharmacology, Jagiellonian University Medical College, Cracow, Poland.
authors.forename: S
authors.initials: S
authors.lastname: Chlopicki
authors.affiliation: Centre for Nanometer-Scale Science and Advanced Materials (NANOSAM) Faculty of Physics, Astronomy and Applied Computer Science, Jagiellonian University, Cracow, Poland.
authors.forename: M
authors.initials: M
authors.lastname: Szymonski
mesh_terms: D000818:Animals; D002196:Capillaries / Q000648:ultrastructure*; D042783:Endothelial Cells / Q000648:ultrastructure*; D008099:Liver / Q000648:ultrastructure*; D051379:Mice; D018625:Microscopy, Atomic Force
keywords: AFM imaging; LSEC; LSEC nanomechanics; fenestrations; fixed cells; force mapping; live cells