NETQUANT: automated quantification of neutrophil extracellular traps. Front Immunol. 8, 1999 (2017). Article PubMed Google Scholar Azzouz, D., Khan, M. A., Sweezey, N. & Palaniyar, N. Two-in-one: UV radiation simultaneously induces apoptosis and NETosis. Cell Death Discov. 4, 51 (...
cell suspension was passed through 70- and 40-μm cell strainers. After RBC lysis, cells were washed and resuspended in staining buffer (1 × PBS/2% FBS). Total and trypan blue-negative (viable) cells were counted with an automated cell counter (BioRad). Representative images of propidium...
In this chapter, we describe an automated high-throughput method to quantify NETting neutrophils in real time using a two-color, live-content imaging platform, the IncuCyteS3 (Essen BioScience, Inc.) system, coupled to membrane integrity-dependent dual-dye approach to image intracellular and ...
Cell-cell interactions maintain tissue homeostasis and contribute to dynamic alteration of the tumor microenvironment (TME). Communication between cancer and host cells not only promotes advanced disease aggression but also determines therapeutic respons
Densitometry was done using the automated digitizing software (Image J, NIH, Bethesda, USA). All phosphorylation levels were normalized to the protein expression of β-actin. Statistical analysis Between-group differences were examined for statistical significance using Student’s t-test. P-values of...
4 PCR amplification was performed in an automated thermal cycler (Bio Phase; Bio Excellence, Colchester, UK) using denaturation for 2 minutes, primer annealing at 55°C, and extension at 72°C for 2 minutes (30 cycles). Twenty microliters of PCR products was fractionated by agarose gel ...
Automated sequence analysis was performed providing unambiguous assignment of the primary sequence of BDP-13, YKIIQQWPHYRRV (Fig. 1, double-underlined sequence). As discussed below, the sequence was confirmed by comparison with the bOBP cDNA. Synthetic BDP-13 was used to produce anti-bOBP ...
not always purified at very high levels (<97%); they are therefore often contaminated by mononuclear cells, which may produce confounding results, especially in terms of gene expression and cytokine release.46Similarly, neutrophils must be carefully and rapidly processed in lipopolysaccharide (LPS)-fr...
Here, we report a haptotaxis assay, based on the classic under-agarose assay, which combines an optical patterning technique to generate surface-bound formyl peptide gradients as well as an automated imaging and analysis of a large number of migration trajectories. We show that human neutrophils ...
The derived parameters can be used to generate histograms, scattergrams, and/or other graphical displays in which the EGC and neutrophil populations are separated sufficiently such that automated enumeration of the respective populations can be achieved. ...