Intravital Imaging

Liver Reticular Fiber Network:

Injection of a fluorescent probe (blue) labels the reticular fiber network of the live liver. Kupffer cells stained with F4/80 (pink) and autofluorescent hepatocytes (green).

Neutrophils and NETs in Human Inflammation: Immunofluoresence labeling of human inflammatory infiltrate obtained from an abscess. Leukocytes (blue), neutrophils (red), DNA (green).

Neutrophils and NETs in Human Inflammation:

Immunofluoresence labeling of human inflammatory infiltrate obtained from an abscess. Leukocytes (blue), neutrophils (red), DNA (green).

Immunofluorescence staining of a human blood smear:

A single neutrophil is labelled by anti-CD66b (red) and DNA labelled by Sytox Green (green). Red blood cells appear autofluorescence (blue).

Imaging of the tumour "landscape":

Intravital imaging of a CD26 tumour following oncolytic viral treatment. The tumour expresses a fluorescent reporter protein (red) and infected cells express a virus-delivered GFP gene (bright green). Various leukocyte populations are labelled in blue, magenta and orange.

Neutrophils and NETs in Human Inflammation:

Immunofluoresence labeling of human inflammatory infiltrate obtained from an abscess. Leukocytes (blue), neutrophils (red), DNA (green).

Multiple leukocyte populations in the spleen:

Intravital imaging of the spleen revealing the presence of B cells (blue), neutrophils (orange), T cells (green) and macrophage (red).

Intravital imaging of the lung:

Neutrophils have been stained with anti-Ly6G (green), monocytes with anti-CD11b (blue) and the vasculature labelled with a contrast agent (red). Circular black areas correspond to the air spaces in the lung.

3D-Image Reconstruction of Neutrophil-Platelet Aggregates in the Liver Following Acute Viral Infection :

3D images rendered from z-stacks captured during intravital imaging of the liver in an uninfected, control mouse (top) and in a mouse infected with Myxoma virus (bottom). Platelets (red) can be observed forming large aggregates, wrapping around the surface of adherent neutrophils (blue) within the liver vasculature. Virally infected cells are shown in green.

Deposition of Neutrophil Extracellular Traps (NETs) in the Liver Vasculature Following Viral Challenge:

Staining for extracellular histone (red), a hallmark feature of NETs, reveals punctate staining along the sinusoidal walls of the liver following challenge with a viral analogue. Neutrophils within the liver vasculature are shown in blue.

Platelet Aggregation Within The Liver Following Viral Challenge:

Platelets (red) binding to, and aggregating on adherent neutrophils (blue) within the liver vasculature following systemic challenge with a viral analogue. Hepatocytes are autofluorescent (green).

Acute Virus Infection Of The Liver Induces A Potent Inflammatory Response:

Infection of the liver with Myxoma virus (green) induces a robust innate immune response including massive neutrophil (blue) and platelet (red) recruitment to the liver vasculature.

Platelet Aggregation Within The Liver Following Viral Challenge:

Platelets (red) can be observed binding to, and aggregating on adherent neutrophils (blue) within the liver vasculature following systemic challenge with a viral analogue.

Videos

Intravital imaging of leukocytes rolling along the vasculature of a virus challenged animal. Neutrophils are labelled with anti-Ly6G (yellow), platelets (red) and virus (blue).

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Intravital imaging of neutrophil and platelet recruitment to the liver following viral challenge. Neutrophils are labelled with anti-Ly6G (blue), platelets (red).

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Resonant-scanning intravital imaging of neutrophils and platelets within the liver vasculature. Neutrophils are labelled with anti-Ly6G (blue), platelets (red) and blood flow is visualized with the aid of a contrast agent (grey).

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4D Resonant-scanning intravital imaging of recruited neutrophils within the liver of a virus challenged mouse. Neutrophils are labelled with anti-Ly6G (purple), and the liver is visualized due to the autofluoresence of the hepatocytes (green).

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4D intravital imaging of recruited neutrophils and platelet aggregation within the liver of a virus challenged mouse. 3D isosurface models of labelled cells are generated through the assembly of z-stacks of focal planes. Neutrophils are labelled with anti-Ly6G (blue), and platelets are labelled with anti-CD49b (red).

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Resonant-scanning intravital imaging of recruited neutrophils and platelets within the lung of a virus challenged mouse. Neutrophils have been stained with anti-Ly6G (purple), platelets with anti-CD49b (red). Blood flow is visible due to the use of a contrast agent (green). Circular black areas correspond to the air spaces in the lung.

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4D Resonant-scanning intravital imaging of recruited neutrophils within the lung of a virus challenged mouse. Neutrophils have been stained with anti-Ly6G (blue) and the blood flow is visible due to the use of a contrast agent (green). Circular black areas correspond to the air spaces in the lung.

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