Morphology and force probing of primary murine liver sinusoidal endothelial cells

• Published in: Journal of molecular recognition 2017-07, Vol.30 (7), p.n/a
• Main Authors: Zapotoczny, B., Owczarczyk, K., Szafranska, K., Kus, E., Chlopicki, S., Szymonski, M.
• Format: Article
• Language: English
• Subjects: AFM imaging; Animals; Atomic force microscopy; Atomic structure; Capillaries - ultrastructure; Endothelial cells; Endothelial Cells - ultrastructure; fenestrations; fixed cells; force mapping; Genes; Glutaraldehyde; High resolution; Homeostasis; Image resolution; Imaging; live cells; Liver; Liver - ultrastructure; LSEC; LSEC nanomechanics; Mice; Microscopy; Microscopy, Atomic Force; Modulation; Morphology; Nanostructure; Nuclei (cytology); Nuclei (nuclear physics); Plates; Porosity; Sieve plates; Visualization
• ISSN: 0952-3499; 1099-1352
• DOI: 10.1002/jmr.2610

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. In this work, we apply atomic force microscopy contact mode to characterize fenestrations in liver sinusoidal endothelial cells (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 cell nuclei in fixed LSECs.