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Microfluidics‐based side view flow chamber reveals tether‐to‐sling transition in rolling neutrophils
Author(s) -
Marki Alex,
Gutierrez Edgar,
Zbigniew Mikulski,
Groisman Alex,
Ley Klaus
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.lb781
Subject(s) - sling (weapon) , shear stress , chemistry , microfluidics , materials science , nanotechnology , composite material , structural engineering , engineering
Neutrophils rolling at high shear stress (above 6 dyn/cm 2 ) form tethers in the rear and slings in front. Here, we developed a novel photo‐lithographically fabricated, PDMS‐based side‐view flow chamber to dynamically visualize tether and sling formation. Fluorescently membrane‐labeled mouse neutrophils rolled on P‐selectin substrate at 10 dyn/cm 2 . Most rolling cells formed 5 tethers that were 2–30 μm long. Breaking of a single tether caused a reproducible forward microjump of the cell, showing that the tether was load‐bearing. About 15% of all tether‐breaking events resulted in slings. The tether‐to‐sling transition was fast (<100 ms) with no visible material extending above the rolling cell, suggesting a very low bending modulus of the tether. The sling extended downstream in front of the rolling cell at a downward angle of 6°, without touching the flow chamber for 3.19±0.78(SEM) sec. These new observations explain how slings form from tethers and provide unexpected insight into their biomechanical properties. Support or Funding Information This project was supported by the P01 HL078784 grant of Klaus Ley.