title: Contribution of high-resolution correlative imaging techniques in the study of the liver sieve in three-dimensions.
abstract: Correlative microscopy has become increasingly important for the analysis of the structure, function, and dynamics of cells. This is largely due to the result of recent advances in light-, probe-, laser- and various electron microscopy techniques that facilitate three-dimensional studies. Furthermore, the improved understanding in the past decade of imaging cell compartments in the third dimension has resulted largely from the availability of powerful computers, fast high-resolution CCD cameras, specifically developed imaging analysis software, and various probes designed for labeling living and or fixed cells. In this paper, we review different correlative high-resolution imaging methodologies and how these microscopy techniques facilitated the accumulation of new insights in the morpho-functional and structural organization of the hepatic sieve. Various aspects of hepatic endothelial fenestrae regarding their structure, origin, dynamics, and formation will be explored throughout this paper by comparing the results of confocal laser scanning-, correlative fluorescence and scanning electron-, atomic force-, and whole-mount electron microscopy. Furthermore, the recent advances of vitrifying cells with the vitrobot in combination with the glove box for the preparation of cells for cryo-electron microscopic investigation will be discussed. Finally, the first transmission electron tomography data of the liver sieve in three-dimensions are presented. The obtained data unambiguously show the involvement of special domains in the de novo formation and disappearance of hepatic fenestrae, and focuses future research into the (supra)molecular structure of the fenestrae-forming center, defenestration center and fenestrae-, and sieve plate cytoskeleton ring by using advanced cryo-electron tomography.
authors.affiliation: Australian Key Centre for Microscopy and Microanalysis, The University of Sydney, New South Wales, Australia. filip.braet@emu.usyd.edu.au
authors.forename: Filip
authors.initials: F
authors.lastname: Braet
authors.forename: Eddie
authors.initials: E
authors.lastname: Wisse
authors.forename: Paul
authors.initials: P
authors.lastname: Bomans
authors.forename: Peter
authors.initials: P
authors.lastname: Frederik
authors.forename: Willie
authors.initials: W
authors.lastname: Geerts
authors.forename: Abraham
authors.initials: A
authors.lastname: Koster
authors.forename: Lilian
authors.initials: L
authors.lastname: Soon
authors.forename: Simon
authors.initials: S
authors.lastname: Ringer
mesh_terms: D000818:Animals; D020285:Cryoelectron Microscopy / Q000379:methods; D022781:Hepatocytes / Q000166:cytology* / Q000648:ultrastructure*; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000379:methods*; D018625:Microscopy, Atomic Force / Q000379:methods; D008855:Microscopy, Electron, Scanning / Q000379:methods; D046529:Microscopy, Electron, Transmission / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods