학술논문

Actin-based protrusions of migrating neutrophils are intrinsically lamellar and facilitate direction changes.
Document Type
article
Source
Subject
Neutrophils
HL-60 Cells
Pseudopodia
Humans
Actins
Microscopy
Cell Movement
Time-Lapse Imaging
actin
amoeboid motility
arp2/3
cell biology
cell migration
human
microscopy
visualization
Biochemistry and Cell Biology
Language
Abstract
Leukocytes and other amoeboid cells change shape as they move, forming highly dynamic, actin-filled pseudopods. Although we understand much about the architecture and dynamics of thin lamellipodia made by slow-moving cells on flat surfaces, conventional light microscopy lacks the spatial and temporal resolution required to track complex pseudopods of cells moving in three dimensions. We therefore employed lattice light sheet microscopy to perform three-dimensional, time-lapse imaging of neutrophil-like HL-60 cells crawling through collagen matrices. To analyze three-dimensional pseudopods we: (i) developed fluorescent probe combinations that distinguish cortical actin from dynamic, pseudopod-forming actin networks, and (ii) adapted molecular visualization tools from structural biology to render and analyze complex cell surfaces. Surprisingly, three-dimensional pseudopods turn out to be composed of thin (