Visualization of membrane rafts and fenestrations.

<p>(A–C) 3D-SIM of LSECs stained with Bodipy FL C5 ganglioside GM1, a marker for rafts (green) and Cell-Mask Orange, a cell membrane marker (orange). There is an inverse distribution between liver sieve plates and membrane rafts. Membrane rafts are mostly perinuclear while sieve plates are mostly peripheral. Some sieve plates are identified by an asterix (*) and fenestrations can be resolved within the sieve plates. Rafts are shown with arrows (→). The areas marked in a box (clustered rafts) are further magnified in (D–G). (D–G) Magnification of areas in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0046134#pone-0046134-g001" target="_blank">Figure 1(A–B)</a> showing clustered membrane rafts with raised perimeters. (H) TIRFM of LSEC stained with NBD-cholesterol (green), a marker for rafts, and CellMask Orange (orange) showing perinuclear distribution of rafts (arrows). Fenestrations are not resolved within the sieve plates (*) with TIRFM. (I) TIRFM of LSEC stained with Bodipy FL C5 ganglioside GM1, a marker for rafts, and CellMask Orange (orange) confirming mostly perinuclear distribution of rafts. Scale bar 5 µm (A, B, H, I), 1 µm (C–G).