Category Archives: Biomaterials

Congratulations Joe Lopes – CREST fellow spring 2017

Congratulations go to Joe, who was selected as one of just four UC Merced graduate students to recieve funding for the Spring semester as a CREST fellow.

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The CREST center (Center for cellular and biomolecular machines) is funded by the National Science Foundation and you can read more about their work and opportunities here.

Joe is working on kinesin – based  microtubule transport and active matter.

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Microtubules gliding on kinesin motors

 

 

 

 

Studying microtubule spools

Understanding the role of transport velocity in biomotor-powered microtubule spool assembly

Amanda J. Tan, Dail E. Chapman, Linda S. Hirst and Jing Xu

In this new paper we examined the sensitivity of microtubule spools to transport velocity.
Perhaps surprisingly, we determined that the steady-state
number and size of spools remained constant over a seven-fold
range of velocities. Our data on the kinetics of spool assembly
further suggest that the main mechanisms underlying spool growth
vary during assembly.
Read the paper in RSC Advances here

Using molecular motors to extract lipid tubules

A Simple Experimental Model to Investigate Force Range for Membrane Nanotube Formation

Chai Lor1, Joseph D. Lopes2, Michelle K. Mattson-Hoss3, Jing Xu2* and Linda S. Hirst2*
  • 1Biological Engineering and Small-scale Technologies Graduate Program, School of Engineering, University of California Merced, Merced, CA, USA
  • 2Physics Department, School of Natural Science, University of California Merced, Merced, CA, USA
  • 3Developmental and Cell Biology, School of Biological Sciences, University of California Irvine, Irvine, CA, USA

fmats-03-00006-g001The presence of membrane tubules in living cells is essential to many biological processes. In cells, one mechanism to form nanosized lipid tubules is via molecular motor induced bilayer extraction. In this paper, we describe a simple experimental model to investigate the forces required for lipid tube formation using kinesin motors anchored to 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) vesicles. Previous related studies have used molecular motors actively pulling on the membrane to extract a nanotube. Here, we invert the system geometry; molecular motors are used as static anchors linking DOPC vesicles to a two-dimensional microtubule network and an external flow is introduced to generate nanotubes facilitated by the drag force. We found that a drag force of ≈7 pN was sufficient for tubule extraction for vesicles ranging from 1 to 2 μm in radius. By our method, we found that the force generated by a single molecular motor was sufficient for membrane tubule extraction from a spherical lipid vesicle.

Congratulations Dr Lor!

Dr Chai Lor successfully defended his PhD thesis in the BEST (Bioengineering and small scale technologies) program on October 26th. Chai was a bioengineering undergraduate at UC Merced and a member of the first graduating class.

“Phase Behavior and Nanotube Formation in Lipid Membranes”

Biological cells are protected by a complex barrier called the lipid membrane. The lipid membrane is a soft material structure consisting of many lipid molecules held together by hydrophobic forces in an aqueous solution. Two simple experimental models were employed to investigate the role of specific lipid molecules in biological membranes. The first model investigated the phase behavior of the binary lipid mixture, 1-dipalmitoyl-2-docosahexaenoyl-sn-glycero-3-phosphoethanolamine (DHA-PE) and 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), using small-angle x-ray scattering (SAXS) and wide-angle x-ray scattering (WAXS). Our results shows that DHA-PE induces phase separation into a DHA rich liquid crystalline (Lα) phase and a DHA poor gel (Lβ’) phase at overall DHA-PE concentrations as low as 0.1mol%. In addition, we find that the structure of the Lβ’ phase, from which the DHA-PE molecules are largely excluded, is modified in the phase-separated state at low DHA-PE concentrations, with a decrease in bilayer thickness of 1.34nm for 0.1mol% at room temperature compared to pure DPPC bilayers. The second model investigated the formation of lipid nanotubes using an anchor system consisting of lipids, kinesin molecular motors, and microtubules in a flow cell. Lipid tubulation was conducted on two different membranes, 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and DPPC. Lipid nanotubes were pulled from anchored giant unilamellar vesicles (GUVs) by drag force generated from the flow inside the channel. The results showed that DPPC membranes cannot generate lipid nanotubes while DOPC can, which was expected. We find that a drag force of approximately ≈7.9 pN is sufficient for tubule extraction and that it only requires 1-2 kinesin motor proteins for anchoring the GUV.

“Low concentrations of docosahexanoic acid significantly modify membrane structure and phase behavior”
C. Lor and L.S. Hirst, MEMBRANES, 5(4), 857-874 Link (2015)

 

Soft particle trapping in a dual-beam optical trap

Stability and instability for low refractive-index-contrast particle trapping in a dual-beam optical trap

Alison Huff, Charles N. Melton, Linda S. Hirst, and Jay E. Sharping

Biomedical optics express, 6 (10), 3812-3819 (2015)

A dual-beam optical trap is used to trap and manipulate dielectric particles. When the refractive index of these particles is comparable to that of the surrounding medium, equilibrium trapping locations within the system shift from stable to unstable depending on fiber separation and particle size. This is due to to the relationship between gradient and scattering forces. We experimentally and computationally study the transitions between stable and unstable trapping of poly(methyl methacrylate) beads for a range of parameters relevant to experimental setups involving giant unilamellar vesicles. We present stability maps for various fiber separations and particle sizes, and find that careful attention to particle size and configuration is necessary to obtain reproducible quantitative results for soft matter stretching experiments.

Read the full paper here

Bifurcation plots showing the axial stability locations vs. particle radius for three different traps with fiber separations of (a) 45 μm, (b) 92.9 μm, and (c) 129.1 μm. The red dots represent the simulated stable trapping location, and the symbols represent experimentally observed stably trapped particles. The trap center is located at z=0. Trapping locations are shown schematically within (c). The error bars were determined empirically through measurements of the inner and outer edges of both the fiber and beads.
Bifurcation plots showing the axial stability locations vs. particle radius for three different traps with fiber separations of (a) 45 μm, (b) 92.9 μm, and (c) 129.1 μm. The red dots represent the simulated stable trapping location, and the symbols represent experimentally observed stably trapped particles. The trap center is located at z=0. Trapping locations are shown schematically within (c). The error bars were determined empirically through measurements of the inner and outer edges of both the fiber and beads.