title: A standardized method for the analysis of liver sinusoidal endothelial cells and their fenestrations by scanning electron microscopy.
abstract: Liver sinusoidal endothelial cells are the gateway to the liver, their transcellular fenestrations allow the unimpeded transfer of small and dissolved substances from the blood into the liver parenchyma for metabolism and processing. Fenestrations are dynamic structures--both their size and/or number can be altered in response to various physiological states, drugs, and disease, making them an important target for modulation. An understanding of how LSEC morphology is influenced by various disease, toxic, and physiological states and how these changes impact on liver function requires accurate measurement of the size and number of fenestrations. In this paper, we describe scanning electron microscopy fixation and processing techniques used in our laboratory to ensure reproducible specimen preparation and accurate interpretation. The methods include perfusion fixation, secondary fixation and dehydration, preparation for the scanning electron microscope and analysis. Finally, we provide a step by step method for standardized image analysis which will benefit all researchers in the field.
authors.affiliation: Centre for Education and Research on Ageing & ANZAC Research Institute, University of Sydney and Concord Hospital; Ageing and Alzheimers Institute, Concord Hospital; Charles Perkins Centre, University of Sydney; victoria.cogger@sydney.edu.au.
authors.forename: Victoria C
authors.initials: VC
authors.lastname: Cogger
authors.affiliation: Centre for Education and Research on Ageing & ANZAC Research Institute, University of Sydney and Concord Hospital; Ageing and Alzheimers Institute, Concord Hospital.
authors.forename: Jennifer N
authors.initials: JN
authors.lastname: O'Reilly
authors.affiliation: Centre for Education and Research on Ageing & ANZAC Research Institute, University of Sydney and Concord Hospital; Ageing and Alzheimers Institute, Concord Hospital; Charles Perkins Centre, University of Sydney.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Warren
authors.affiliation: Centre for Education and Research on Ageing & ANZAC Research Institute, University of Sydney and Concord Hospital; Ageing and Alzheimers Institute, Concord Hospital; Charles Perkins Centre, University of Sydney.
authors.forename: David G
authors.initials: DG
authors.lastname: Le Couteur
mesh_terms: D000818:Animals; D042783:Endothelial Cells / Q000166:cytology / Q000648:ultrastructure*; D022781:Hepatocytes / Q000166:cytology; D007091:Image Processing, Computer-Assisted / Q000379:methods; D008099:Liver / Q000166:cytology* / Q000648:ultrastructure; D051379:Mice; D008855:Microscopy, Electron, Scanning / Q000379:methods*; D051381:Rats