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Image Search Results
Journal: Bio-protocol
Article Title: In vitro Demonstration and Quantification of Neutrophil Extracellular Trap Formation
doi: 10.21769/BioProtoc.2386
Figure Lengend Snippet: A. Human neutrophils were isolated from fresh peripheral blood of healthy donors by gradient centrifugation (step A1). The neutrophil layer at the interface of H1077 and H1119 were collected. PBMC, peripheral blood mononuclear cells; PMN, polymorphonuclear leukocytes; RBC, red blood cells. B. The cell viability was analyzed with PI dye and the purity of the viable neutrophils was analyzed using a human CD15 antibody within the PI negative gate. C. Human neutrophils were isolated from buffy coat with the MACSxpress method (step A2). The cell viability was analyzed with PI dye and the purity of the viable neutrophils was analyzed with human CD15 antibody within the PI negative gate. D. Murine bone marrow neutrophils were isolated by magnetic sorting using a negative depletion method (step A3). The cell viability was analyzed with PI dye and the purity of the viable neutrophils was analyzed with a mouse Ly6G (Gr-1) antibody within the PI negative gate. Gray histograms: PI dye or CD15 antibody or Gr-1 antibody; open histogram: unstained control or isotype antibody.
Article Snippet: Isolate neutrophils by immunomagnetic separation (MACS) using the negative depletion method with
Techniques: Isolation, Gradient Centrifugation, Control
Journal: Bio-protocol
Article Title: In vitro Demonstration and Quantification of Neutrophil Extracellular Trap Formation
doi: 10.21769/BioProtoc.2386
Figure Lengend Snippet: 2 x 105 human neutrophils (A) or murine neutrophils (B) were activated by 100 ng/ml PMA for 3 h in wells of glass chamber slide. Resting neutrophils with DMSO served as negative control. Slides were fixed and labelled with anti-DNA/Histone H1 antibody followed by AF555-conjugated anti-mouse IgG (red) and nuclei were counterstained with DAPI (blue) (steps B1 and B2). C. Immunostaining of human neutrophil marker CD15 (green) and NET marker DNA/Histone H1 (red) on paraffin-embedded skin section from human vasculitis patients. D. Immunostaining of the murine neutrophil marker Ly6G (green) and NET marker DNA/Histone H1 (red) on paraffin-embedded skin section from mice with induced immune complex-mediated vasculitis (step B3). Scale bars = 100 µm.
Article Snippet: Isolate neutrophils by immunomagnetic separation (MACS) using the negative depletion method with
Techniques: Negative Control, Immunostaining, Marker