Figure 1.
SD-IVM of the liver allows one to pinpoint the location of platelet clearance and to analyze platelet Kupffer cell interactions in the steady state.(A) Schematic depiction of the liver structure (left) and example image of the mouse liver (right) show that crucial components of the liver can be stained and visualized using SD-IVM. Kupffer cells and endothelial cells were stained using fluorescently labeled anti-F4/80 and anti-CD31 antibody, respectively. LSEC, liver sinusoidal endothelial cell; Ø, diameter. Scale bar is 70 µm. (B) 3D reconstruction of a 15-µm z-stack obtained using SD-IVM of the mouse liver 24 h after injecting rhodamine dextran shows colocalization of dextran staining and hepatocytes (overlay of red dextran with green hepatocytes appears yellow). Endothelial cells were stained using fluorescently labeled anti-CD31 antibody. Scale bars are 100 µm. (C) SD-IVM of the mouse liver 24 h after injection of fluorescently labeled albumin shows dotted structure colocalizing with endothelial staining. Kupffer cells and endothelial cells were stained using fluorescently labeled anti-F4/80 and anti-CD31 antibody, respectively. Scale bars are 33 µm. (D) 1,704 interactions of platelets with 124 Kupffer cells were tracked over 30 min using Imaris software, and the dwell time was plotted. (E) Average platelet interactions per Kupffer cell per 30 min, grouped by dwell time. Data represent means ± SEM. (F) Interactions of platelets with a single Kupffer cell are visualized using SD-IVM of the mouse liver. While most interactions are transient, a small proportion (arrows) forms firm interactions. Kupffer cells and endogenous platelets were stained using fluorescently labeled anti-F4/80 and anti-CD49b antibody, respectively. Scale bars are 8 µm. (G) 3D reconstruction of a single Kupffer cell of naive CD41-YFPki/+ mice based on a z-stack obtained using SD-IVM of the liver. The Kupffer cell was labeled using an anti-F4/80 antibody. Note the YFP platelet remnant (bright green) inside the Kupffer cell (arrows). Scale bars are 100 µm. (H) Percentage of Kupffer cells (KCs) in CD41-YFPki/+ mice that harbor platelet-YFP remnants inside. Results from all experiments shown are representative of three independent experiments.

SD-IVM of the liver allows one to pinpoint the location of platelet clearance and to analyze platelet Kupffer cell interactions in the steady state. (A) Schematic depiction of the liver structure (left) and example image of the mouse liver (right) show that crucial components of the liver can be stained and visualized using SD-IVM. Kupffer cells and endothelial cells were stained using fluorescently labeled anti-F4/80 and anti-CD31 antibody, respectively. LSEC, liver sinusoidal endothelial cell; Ø, diameter. Scale bar is 70 µm. (B) 3D reconstruction of a 15-µm z-stack obtained using SD-IVM of the mouse liver 24 h after injecting rhodamine dextran shows colocalization of dextran staining and hepatocytes (overlay of red dextran with green hepatocytes appears yellow). Endothelial cells were stained using fluorescently labeled anti-CD31 antibody. Scale bars are 100 µm. (C) SD-IVM of the mouse liver 24 h after injection of fluorescently labeled albumin shows dotted structure colocalizing with endothelial staining. Kupffer cells and endothelial cells were stained using fluorescently labeled anti-F4/80 and anti-CD31 antibody, respectively. Scale bars are 33 µm. (D) 1,704 interactions of platelets with 124 Kupffer cells were tracked over 30 min using Imaris software, and the dwell time was plotted. (E) Average platelet interactions per Kupffer cell per 30 min, grouped by dwell time. Data represent means ± SEM. (F) Interactions of platelets with a single Kupffer cell are visualized using SD-IVM of the mouse liver. While most interactions are transient, a small proportion (arrows) forms firm interactions. Kupffer cells and endogenous platelets were stained using fluorescently labeled anti-F4/80 and anti-CD49b antibody, respectively. Scale bars are 8 µm. (G) 3D reconstruction of a single Kupffer cell of naive CD41-YFPki/+ mice based on a z-stack obtained using SD-IVM of the liver. The Kupffer cell was labeled using an anti-F4/80 antibody. Note the YFP platelet remnant (bright green) inside the Kupffer cell (arrows). Scale bars are 100 µm. (H) Percentage of Kupffer cells (KCs) in CD41-YFPki/+ mice that harbor platelet-YFP remnants inside. Results from all experiments shown are representative of three independent experiments.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal