Computational protocol: Beware the intruder: Real time observation of infiltrated neutrophils and neutrophil—Microglia interaction during stroke in vivo

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Protocol publication

[…] The fluorescence staining with fluorescein coupled solanum tuberosum lectin was performed in 50 μm thick frozen sections from bone marrow chimeric mice (C57BL6 reconstituted with bone marrow from CatchupIVM mice). The brains were fixed with 4% paraformaldehyde 24 h after photothrombosis by transcardial perfusion. Microscopic pictures were done with a Leica SP8 (Wetzlar, Germany) confocal microscope (objective: 20x water-immersion, excitation of FITC: 488 nm, CatchupIVM: 561 nm). IMARIS (Bitplane, Zurich) was used for preparing the movies and images. [...] IMARIS (Bitplane, Zurich) was used for automated tracking (velocity) and sphericity measurements of neutrophils. Statistical analysis were performed using GraphPadPrism (LA Jolla, USA). For comparison of two groups unpaired two-sided Student t-tests were applied. A value of p<0.05 was considered statistically significant. […]

Pipeline specifications

Software tools Imaris, GraphPad Prism
Applications Miscellaneous, Laser scanning microscopy
Organisms Mus musculus
Diseases Cerebral Infarction, Stroke