| Direct Measures of Large, Anisotropic Strains in Deformation of the Erythrocyte Cytoskeleton Biophysical Journal, Volume 77, Issue 2, 1 August 1999, Pages 853-864 James C-M. Lee, Derek T. Wong and Dennis E. Discher Abstract The erythrocyte's spectrin–actin membrane skeleton is directly shown to be capable of sustaining large, anisotropic strains. Photobleaching of an ∼1-m stripe in rhodamine phalloidin-labeled actin appears stable up to at least 37°C, and is used to demonstrate large in-surface stretching during elastic deformation of the skeleton. Principal extension or stretch ratios of at least ∼200% and contractions down to ∼40%, both referenced to an essentially undistorted cell, are visually demonstrated in micropipette-imposed deformation. Such anisotropic straining is seen to be consistent at a qualitative level with now classic analyses (Evans. 1973. 13:941–954) and is generally nonhomogeneous though axisymmetric down to the submicron scale. Local, direct measurements of stretching prove quantitatively consistent (within ∼10%) with integrated estimates that are based simply on a measured relative density distribution of actin. The measurements are also in close agreement with direct computation of mean spectrin chain extension in full statistical mechanical simulations of a coarse-grained network held in a micropipette. Finally, as a cell thermally fragments near ∼48°C, the patterned photobleaching demonstrates a destructuring of the surface network in a process that is more readily attributable to transitions in spectrin than in F-actin. Abstract | Full Text | PDF (527 kb) |
| Actin Protofilament Orientation in Deformation of the Erythrocyte Membrane Skeleton Biophysical Journal, Volume 79, Issue 6, 1 December 2000, Pages 2987-3000 Catherine Picart, Paul Dalhaimer and Dennis E. Discher Abstract The red cell’s spectrin-actin network is known to sustain local states of shear, dilation, and condensation, and yet the short actin filaments are found to maintain membrane-tangent and near-random azimuthal orientations. When calibrated with polarization results for single actin filaments, imaging of micropipette-deformed red cell ghosts has allowed an assessment of actin orientations and possible reorientations in the network. At the hemispherical cap of the aspirated projection, where the network can be dilated severalfold, filaments have the same membrane-tangent orientation as on a relatively unstrained portion of membrane. Likewise, over the length of the network projection pulled into the micropipette, where the network is strongly sheared in axial extension and circumferential contraction, actin maintains its tangent orientation and is only very weakly aligned with network extension. Similar results are found for the integral membrane protein Band 3. Allowing for thermal fluctuations, we deduce a bound for the effective coupling constant, , between network shear and azimuthal orientation of the protofilament. The finding that must be about an order of magnitude or more below its tight-coupling value illustrates how nanostructural kinematics can decouple from more macroscopic responses. Monte Carlo simulations of spectrin-actin networks at ∼10-nm resolution further support this conclusion and substantiate an image of protofilaments as elements of a high-temperature spin glass. Abstract | Full Text | PDF (813 kb) |
| Double-Tether Extraction from Human Umbilical Vein and Dermal Microvascular Endothelial Cells Biophysical Journal, Volume 92, Issue 3, 1 February 2007, Pages 1035-1045 Gaurav Girdhar, Yong Chen and Jin-Yu Shao Abstract Multiple tethers are very likely extracted when leukocytes roll on the endothelium under high shear stress. Endothelial cells have been predicted to contribute more significantly to simultaneous tethers and thus to the overall rolling stabilization. We therefore extracted and quantified double tethers from endothelial cells with the micropipette aspiration technique. We show that the constitutive parameters (threshold force () and effective viscosity ()) for double-tether extraction are twice those for single-tether extraction and are remarkably similar for human neonatal (=105±5pN; =1.0±0.1pN·s/m) and adult (=118±13pN; =1.3±0.2pN·s/m) dermal microvascular, and human umbilical vein (=99±3pN; =1.0±0.1pN·s/m) endothelial cells. Additionally, these parameters are also independent of surface receptor type, cytokine stimulation, and attachment state of the endothelial cell. We also introduce a novel correlation between the cell-substrate contact stress and gap width, with which we can predict the apparent cell-substrate separation range to be 0.01–0.1m during leukocyte rolling. With a biomechanical model of leukocyte rolling, we calculate the force history on the receptor-ligand bond during tether extraction and predict maximum stabilization for the double simultaneous tether extraction case. Abstract | Full Text | PDF (501 kb) |
Copyright © 1973 The Biophysical Society. All rights reserved.
Biophysical Journal, Volume 13, Issue 9, 941-954, 1 September 1973
doi:10.1016/S0006-3495(73)86036-9
Articles
E.A. Evans
A two-dimensional elastomer material concept of the red cell membrane is applied to the analysis of fluid shear-deformed, point-attached red cells and micropipette aspiration of red cell disks. The elastic constant (corresponding to the “shear” modulus multiplied by the membrane thickness) is of the order 10-2 dyn/cm for both cases. Additional experimental observations are in agreement with the membrane model, e.g. teardrop and “tether” formation of the sheared disks, pressure difference vs. aspirated length of the cell for micropipette experiments, etc