Structure-function relationship of biological gels revealed by multiple-particle tracking and differential interference contrast microscopy: the case of human lamin networks.
Lamin B1 filaments organize into a thin dense meshwork underlying the nucleoplasmic side of the nuclear envelope. Recent experiments in vivo suggest that lamin B1 plays a key structural role in the nuclear envelope, but the intrinsic mechanical properties of lamin B1 networks remain unknown. To assess the potential mechanical contribution of lamin B1 in maintaining the integrity and providing structural support to the nucleus, we measured the micromechanical properties and examined the ultrastructural distribution of lamin B1 networks in vitro using particle tracking methods and differential interference contrast (DIC) microscopy. We exploit various surface chemistries of the probe microspheres (carboxylated, polyethylene glycol-coated, and amine-modified) to differentiate lamin-rich from lamin-poor regions and to rigorously extract local viscoelastic moduli from the mean-squared displacements of noninteracting particles. Our results show that human lamin B1 can, even in the absence of auxiliary proteins, form stiff and yet extremely porous networks that are well suited to provide structural strength to the nuclear lamina. Combining DIC microscopy and particle tracking allows us to relate directly the local organization of a material to its local mechanical properties, a general methodology that can be extended to living cells.
Panorchan
Porntula
P
Department of Chemical and Biomolecular Engineering, The Johns Hopkins University, 3400 North Charles Street, Baltimore, Maryland 21218, USA.
Wirtz
Denis
D
Tseng
Yiider
Y
D004058:Diffusion; D004548:Elasticity; D020100:Hydrogels / Q000032:analysis / Q000737:chemistry*; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D034921:Lamin Type B / Q000032:analysis / Q000737:chemistry* / Q000648:ultrastructure*; D008422:Materials Testing / Q000379:methods*; D044085:Microfluidics / Q000379:methods*; D008858:Microscopy, Phase-Contrast / Q000379:methods*; D008863:Microspheres; D015335:Molecular Probes; D009038:Motion; D046912:Multiprotein Complexes / Q000032:analysis / Q000737:chemistry / Q000648:ultrastructure; D013329:Structure-Activity Relationship; D014783:Viscosity
D020100:Hydrogels; D034921:Lamin Type B; D015335:Molecular Probes; D046912:Multiprotein Complexes