Journal: PLoS ONE

Article Title: Entamoeba histolytica Trophozoites and Lipopeptidophosphoglycan Trigger Human Neutrophil Extracellular Traps

doi: 10.1371/journal.pone.0158979

Figure Lengend Snippet: A) Growth kinetic of trophozoites co-incubated with NETs for the indicated times (5 to 60 min) and thereafter cultured in fresh TYI-S-33 medium. Live trophozoites were counted every 24 h under light microscope using Trypan blue. Data shown at each time is the mean ± SD of three independent assays. B) Immunofluorescence assay upon NETs induced by amoebas at 15 and 60 min (left panels) using anti-LL-37 antibody and an anti-rabbit IgG conjugated to FITC (right panels). Magnification 40X.

Article Snippet: All the samples with fluorescence and immunocytochemistry were analyzed with a Carl Zeiss microscope (Axio Imager M1) and the microphotographs were taken with the AxioCam MRc5 digital camera (Carl Zeiss).

Techniques: Incubation, Cell Culture, Light Microscopy, Immunofluorescence

Journal: PLoS ONE

Article Title: Entamoeba histolytica Trophozoites and Lipopeptidophosphoglycan Trigger Human Neutrophil Extracellular Traps

doi: 10.1371/journal.pone.0158979

Figure Lengend Snippet: NETs induced with PMA were incubated 1 h with amoeba trophozoites. Top: A-D, Sytox Green; E and F, Hoechst 33342. A, B at 10X and C-F at 40X objectives at the fluorescence microscope. A and C, phase contrast fields corresponding to B and D, respectively. F, florescence; E, merges phase contrast with fluorescence. Arrow tips, E . histolytica trophozoites. Bottom: after amoeba-NET interaction, trophozoites were growth in TYI-S-33 culture medium during 72 h and cells harvested were counted under a light microscope. Controls, trophozoites cultured at the same conditions in the absence of NETs. Amoeba growth after interaction with NETs and EGTA or EGTA + E-64 was significantly different from controls without NETs (p <0.001). No other significant difference (p <0.05) was observed compared to the control without NETs in the absence of other compounds.

Article Snippet: All the samples with fluorescence and immunocytochemistry were analyzed with a Carl Zeiss microscope (Axio Imager M1) and the microphotographs were taken with the AxioCam MRc5 digital camera (Carl Zeiss).

Techniques: Incubation, Fluorescence, Microscopy, Light Microscopy, Cell Culture, Control

Journal: PLoS ONE

Article Title: Entamoeba histolytica Trophozoites and Lipopeptidophosphoglycan Trigger Human Neutrophil Extracellular Traps

doi: 10.1371/journal.pone.0158979

Figure Lengend Snippet: Human neutrophils were incubated for 2 h at 37°C with 0 to 15 ng/μL of purified EhLPPG. A, C, E and G, phase contrast fields of B, D, F, and H, respectively. Sytox Green observed at 40X objective in the fluorescence microscope.

Article Snippet: All the samples with fluorescence and immunocytochemistry were analyzed with a Carl Zeiss microscope (Axio Imager M1) and the microphotographs were taken with the AxioCam MRc5 digital camera (Carl Zeiss).

Techniques: Incubation, Purification, Fluorescence, Microscopy

Journal: PLoS ONE

Article Title: Entamoeba histolytica Trophozoites and Lipopeptidophosphoglycan Trigger Human Neutrophil Extracellular Traps

doi: 10.1371/journal.pone.0158979

Figure Lengend Snippet: Induction of neutrophil extracellular traps by 10 ng/μL of EhLPPG during the times indicated. Control, neutrophils incubated at the same conditions for 2 h in the absence of EhLPPG. Hoechst 33342 stain and confocal microscope observation, bars 10 μm.

Article Snippet: All the samples with fluorescence and immunocytochemistry were analyzed with a Carl Zeiss microscope (Axio Imager M1) and the microphotographs were taken with the AxioCam MRc5 digital camera (Carl Zeiss).

Techniques: Control, Incubation, Staining, Microscopy

Journal: PLoS ONE

Article Title: Entamoeba histolytica Trophozoites and Lipopeptidophosphoglycan Trigger Human Neutrophil Extracellular Traps

doi: 10.1371/journal.pone.0158979

Figure Lengend Snippet: Neutrophils or NET induced by EhLPPG were stained by an anti-LL-37 antibody and an anti-rabbit IgG conjugated to Alexa594 (red). Control PMN, polymorphonuclear leucocytes with only the secondary antibody. PMN, PMN stained with anti-LL-37 and the anti-rabbit IgG conjugated to Alexa594. NET, PMN incubated with LPPG showing the release of NET and colocalization with LL-37. Hoechst 33342 stain and confocal microscope observation, bars 10 μm.

Article Snippet: All the samples with fluorescence and immunocytochemistry were analyzed with a Carl Zeiss microscope (Axio Imager M1) and the microphotographs were taken with the AxioCam MRc5 digital camera (Carl Zeiss).

Techniques: Staining, Control, Incubation, Microscopy