Digestion of NETs in Paraffin Sections

The digestion of NETs in paraffin section with external DNase was conducted as described previously (15 (link)). Briefly, the digestion was conducted in one slide for 30 minutes at 37°C. A serial cut of the paraffin section was handled and stained in parallel without adding DNase. Samples were recorded using a Leica TCS SP5 AOBS confocal inverted-base fluorescence microscope with an HCX PL APO ×40 0.75–1.25 oil immersion objective The settings were adjusted using isotype control antibodies in separate preparations. Stitched images were combined using Adobe Photoshop CS6.

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de Buhr N., Baumann T., Werlein C., Fingerhut L., Imker R., Meurer M., Götz F., Bronzlik P., Kühnel M.P., Jonigk D.D., Ernst J., Leotescu A., Gabriel M.M., Worthmann H., Lichtinghagen R., Tiede A., von Köckritz-Blickwede M., Falk C.S., Weissenborn K., Schuppner R, & Grosse G.M. (2022). Insights Into Immunothrombotic Mechanisms in Acute Stroke due to Vaccine-Induced Immune Thrombotic Thrombocytopenia. Frontiers in Immunology, 13, 879157.

Publication 2022

TCS SP5 AOBS HCX PL APO ×40

Antibodies isotype Digestion Dnase Fluorescence microscope Immersion Nets Paraffin

Corresponding Organization : University of Veterinary Medicine Hannover, Foundation

Other organizations : Medizinische Hochschule Hannover, German Center for Lung Research

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Variable analysis

independent variables

Digestion of NETs in paraffin section with external DNase

dependent variables

Samples recorded using a Leica TCS SP5 AOBS confocal inverted-base fluorescence microscope

control variables

Serial cut of the paraffin section handled and stained in parallel without adding DNase

controls

Isotype control antibodies in separate preparations

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