celltrace™ far red fluorescent dye  (Thermo Fisher)

Journal: Infection and Immunity

Article Title: Survival of Streptococcus suis in Porcine Blood Is Limited by the Antibody- and Complement-Dependent Oxidative Burst Response of Granulocytes

doi: 10.1128/IAI.00598-19

Figure Lengend Snippet: Influence of complement and NADPH oxidase inhibition on the rate of phagocytosis of porcine granulocytes in blood reconstituted with hyperimmune serum. (A) Blood reconstituted with hyperimmune serum raised against cps2 strain 10 was infected with CellTrace far-red (FR)-labeled S. suis strain 10, and samples were analyzed by flow cytometry to estimate the oxidative burst response (Rho123 signal) within phagocytic granulocytes. (B) Complete phagocytosis rate depicted as a percentage of S. suis FR-positive granulocytes. Serum was untreated, heat inactivated (56°C for 30 min) to inhibit complement, and/or treated with the NADPH oxidase inhibitor apocynin (1.5 mM). Bars and error bars represent means and standard deviations, and significant differences are indicated. Statistical significances were calculated using the Kruskal-Wallis test with Dunn’s multiple-comparison test (n = 7) (*, P < 0.05; ***, P < 0.001).

Article Snippet: In order to be able to read out the oxidative burst and phagocytosis, living S. suis strain 10 bacteria were labeled with CellTrace far-red fluorescent dye (Thermo Fisher Scientific) and added to the blood samples at a concentration of 2 × 10 6 CFU/ml.

Techniques: Inhibition, Infection, Labeling, Flow Cytometry, Comparison

Journal: Nature cell biology

Article Title: Obesity alters the lung myeloid cell landscape to enhance breast cancer metastasis through IL5 and GM-CSF

doi: 10.1038/ncb3578

Figure Lengend Snippet: Obesity enhances lung homing of neutrophils in an IL5-dependent manner. (a) Schematic representation of the adoptive cell transfer experiment. Neutrophils were isolated from BM of WT or ob/ob mice, labelled with fluorescent CellTrace dye (green and red, respectively), mixed 1:1, and then injected via the tail vein (3 × 106 cells) into WT or ob/ob mice ± IL5 neutralizing antibody. Lungs were isolated for flow cytometry analysis after 4 h and 8 h to assess kinetics of neutrophil trafficking. (b) Flow cytometry validation of an equal 1:1 mix of WT (green; 49.3%) and ob/ob (red; 49.7%) donor neutrophils immediately prior to adoptive cell transfer injections. (c) Flow cytometry analysis of lung at 4 h post-adoptive transfer. The vast majority of labelled neutrophils at this time point were from ob/ob donors. n = 5 mice per recipient group, mean ± s.e.m., one-way ANOVA and Dunnett’s multiple comparisons test. (d) Representative flow cytometry plots for the data presented in c. (e) Flow cytometry analysis of fluorescently labelled circulating neutrophils 4 h post-adoptive transfer, demonstrating balanced representation of both red (ob/ob donor) and green (WT donor) cells. n = 5 mice per recipient group; mean ± s.e.m. (f) Flow cytometry analysis of lung at 8 h post-adoptive transfer. Equivalent representation of red and green donor neutrophils was observed at this time point compared with 4 h as in c. n = 5 mice per recipient group, mean ± s.e.m., one-way ANOVA and Dunnett’s multiple comparisons test. (g) Representative flow cytometry plots for the data presented in f. NS, not significant.

Article Snippet: WT neutrophils were labelled with green fluorescent CellTrace dye and ob/ob neutrophils were labelled with far red fluorescent CellTrace dye (Invitrogen).

Techniques: Isolation, Injection, Flow Cytometry, Adoptive Transfer Assay