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A non-invasive imaging for the in vivo tracking of high-speed vesicle transport in mouse neutrophils.

ABSTRACT: Neutrophils play an essential role in the innate immune response. To understand neutrophil activity, the development of a new technique to observe neutrophils in situ is required. Here, we report the development of a non-invasive technique for the in vivo imaging of neutrophils labeled with quantum dots, up to 100 ?m below the skin surface of mice. Upon inflammation neutrophils began to extravasate from blood vessels and locomoted in interstitial space. Most intriguingly, the quantum dots were endocytosed into vesicles in the neutrophils, allowing us to track the vesicles at 12.5 msec/frame with 15-24 nm accuracy. The vesicles containing quantum dots moved as "diffuse-and-go" manner and were transported at higher speed than the in vitro velocity of a molecular motor such as kinesin or dynein. This is the first report in which non-invasive techniques have been used to visualize the internal dynamics of neutrophils.

SUBMITTER: Kikushima K

PROVIDER: S-EPMC3668321 | biostudies-literature | 2013

REPOSITORIES: biostudies-literature

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A non-invasive imaging for the in vivo tracking of high-speed vesicle transport in mouse neutrophils.

Kikushima Kenji K Kita Sayaka S Higuchi Hideo H

Scientific reports 20130101

Neutrophils play an essential role in the innate immune response. To understand neutrophil activity, the development of a new technique to observe neutrophils in situ is required. Here, we report the development of a non-invasive technique for the in vivo imaging of neutrophils labeled with quantum dots, up to 100 μm below the skin surface of mice. Upon inflammation neutrophils began to extravasate from blood vessels and locomoted in interstitial space. Most intriguingly, the quantum dots were e ...[more]

PMID: 23722417

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Project description:Brain-infiltrating monocyte-derived macrophages are one of the key players in the local immune response after stroke. It is now widely accepted that the inflammatory response is not an exclusively destructive process. However, the underlying molecular mechanisms needed for proper regulation still remain to be elucidated. Here, we propose an in vitro labelling strategy for multimodal in vivo observation of macrophage dynamics distinguished from brain-residing microglia response. Prior to intracerebral transplantation into the striatum of recipient mice or systemic administration, monocytes and macrophages, isolated from luciferase-expressing mice, were labelled with superparamagnetic iron oxide particles. Temporo-spatial localization was monitored by magnetic resonance imaging, whereas survival of grafted cells was investigated using bioluminescence imaging. The labelling procedure of the isolated cells did not significantly influence cell characteristics and resulted in detection of as few as 500 labelled cells in vivo. Two weeks after stereotactic transplantation, the luciferase signal was sustained traceable, with approximately 18% of the original luciferase signal detectable for monocytes and about 30% for macrophages. Hypointensity in MRI of the graft appeared unaltered in spatial location. In a therapeutically relevant approach, systemic cell administration after stroke resulted in accumulation mostly in thoracic regions, as could be visualized with BLI. For detection of homing to ischemic brain tissue more cells need to be administered. Nevertheless, during parallel MRI sessions recruitment of i.v. injected cells to the lesion site could be detected by day 2 post stroke as scattered hypointense signal voids. With further increase in sensitivity, our multi-facetted labelling strategy will provide the basis for in vivo tracking and fate specification of tissue-infiltrating macrophages and their distinct role in stroke-related neuro-inflammation.

Dataset Information

A non-invasive imaging for the in vivo tracking of high-speed vesicle transport in mouse neutrophils.

Publications

A non-invasive imaging for the in vivo tracking of high-speed vesicle transport in mouse neutrophils.

Similar Datasets

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