Correlative fluorescence-, and SEM micrographs of control (A-C) and microfilament-disrupted LSECs obtained after 10 min (D-F), 60 min (G-I) and 120 min (J-L) di-h-HALI treatment. Figure set shows simultaneous localization of fluorescent labeled F-actin (red) (left column) in combination with topographic SEM information (green) (middle column) and the merged image (right column) of the same cell. Control LSECs show features similar as those seen in Figs. 1A and 2A: i.e., a well developed filamentous actin cytoskeleton (A) and sieve plates (B). The merge image (C) reveals that the fenestrated areas are clearly interspersed between the actin filaments. Scale bar, 2 μm. After 10 min di-h-HALI treatment (D-F), the images show that the brightly stained F-actin dots matches with the fine globular topographic elevations present on the thin nonfenestrated cytoplasmic arms. Within one hour of di-h-HALI treatment (G-I), the merge image of the fenestrated area reveals that the FFC and the area around is devoid of F-actin. Scale bar, 200 nm. (J-L) Images obtained after 120 min of di-h-HALI treatment. Note that the F-actin dots are localized in the thin nonfenestrated cytoplasmic arms; while the highly fenestrated cytoplasm lacks F-actin. Scale bar, 2 μm.
Braet et al. BMC Cell Biology 2002 3:7 doi:10.1186/1471-2121-3-7