Computational protocol: Rapid Sequestration of Leishmania mexicana by Neutrophils Contributes to the Development of Chronic Lesion

Similar protocols

Protocol publication

[…] Mice were anaesthetized with 80mg/kg Ketamin and 8mg/kg Rompun. The ear pinna was spread with the inner side on top on a custom made stage heated to 37°C and processed as reported previously []. Intravital imaging was performed using a Zeiss LSM 710 NLO microscope equipped with a Cameleon Ultra II Ti:Sapphire 2-Photon laser (Coherent) tuned at 960 nm. Emitted fluorescence during concomitant GFP, YFP and DsRed imaging was read out using the descanned detectors of the microscope imaging at 500–520 nm, 525–555 nm, and 560–650 nm, respectively. Two-color YFP and DsRed imaging was detected with non-descanned detectors at 510–555 nm and 560–650 nm, respectively. Four-dimensional full-scale images were analyzed using the Tracking and Colocalization modules of the Imaris Software (Bitplane, Zurich, Switzerland) and the ImageJ software. Tracking of DCs was performed on 20μm Z-projections, distances to the closest parasite were determined automatically for every DC position of the track. […]

Pipeline specifications

Software tools Imaris, ImageJ
Application Microscopic phenotype analysis
Organisms Leishmania mexicana, Mus musculus, Homo sapiens
Diseases Infection, Neutropenia, Leishmaniasis, Cutaneous