Time-lapse Fluorescence Microscopy of Neutrophil Extracellular Traps or NETs: A Technique to Analyze Post-adhesion Interaction between Cancer Cells and NETs
In this video, we analyze the post-adhesion behavior of cancer cells trapped in neutrophil extracellular traps (NETs) by co-culturing low-density neutrophils and gastric cancer cells. The time-lapse video analysis helps monitor various cellular events and study the role of NETs in pathological conditions.
Protocol
1. Time Lapse Video Analysis of the Trapped Tumor Cells Culture the peritoneal LDNs in a 35 mm round dish coated with poly-L-lysine and incubate for 2 h at 37 ˚C in 5% CO2. Add the green-fluorescent dye for staining the nucleus and chromosomes to visualize NETs at the final concentration of 5 µM. After 1 min of incubation, remove the media and add 2 mL of 0.1% BSA + RPMI 1640. Remove the media and add unstained 1 x 106 MK…
Disclosures
The authors have nothing to disclose.
Materials
SYTOX green nucleic acid stain 5mM solution in DMSO
Time-lapse Fluorescence Microscopy of Neutrophil Extracellular Traps or NETs: A Technique to Analyze Post-adhesion Interaction between Cancer Cells and NETs. J. Vis. Exp. (Pending Publication), e20416, doi: (2023).