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
The 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.
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)
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.
This summer Kyle Kabasaras presented his original research project at the UC Merced undergrad research symposium.
Investigating quantum dot assembly in a cholesteric liquid crystal
An ongoing goal in condensed matter physics is directly controlling the self-assembly of quantum dots (QDs) into specific structures while maintaining their original electronic and optical properties. One method of controlling the self-assembly of QDs is to disperse them within a liquid crystal (LC) medium and apply a variety of thermal stimulations. Recently, our lab developed a method of creating spherical, vesicle-shaped QDs within a nematic LC. Vesicle formation depends on the QD concentration in the LC as well as the LC’s intermolecular dispersion forces and thermal properties. In this project, we investigate the dispersion of CdSe/ZnS (core/shell) QDs in a cholesteric LC (CLC) mediumand predict the QD aggregations to cluster near the LC defects. By varying parameters such as QD concentration and temperature, we exploit the CLC’s sensitive optical and thermal properties. To observe these effects, we apply spectrophotometry, polarized optical microscopy, and fluorescence microscopy. These techniques highlight the aggregation of QDs within the host CLC and identify how LC phase transitions determine where QDaggregates form. This work illustrates the possibility of new LC-based QD devices, and we will continue by exploring the lasing potential of our sample.
Congratulations to graduate student Makiko Quint! Makiko was awarded the UC Merced faculty mentor fellowship for 2015/2016. Her research project, in collaboration with the Ghosh group at UC Merced focuses on liquid crystal nanoparticle composites.
The award assists recipients in acquiring and developing advanced research skills under faculty mentorship and is aimed at increasing the number of students who complete their Ph.D. degree and successfully acquire a faculty appointment.
The fellowship provides one year’s funding for Makiko’s graduate work – congratulations!
“All optical switching of nematic liquid crystal films driven by localized surface plasmons” M.T. Quint, S. Delgado, Z.S. Nuno, L.S. Hirst and S. Ghosh, OPTICS EXPRESS, 23,5, 6888 (2015) Link
We have demonstrated an all-optical technique for reversible in-plane and out-of-plane switching of nematic liquid crystal molecules in few micron thick films. Our method leverages the highly localized electric fields (“hot spots”) and plasmonic heating that are generated in the near-field region of densely packed gold nanoparticle layers optically excited on-resonance with the localized surface plasmon absorption. Using polarized microscopy and transmission measurements, we observe this switching from homeotropic to planar over a temperature range starting at room temperature to just below the isotropic transition, and at on-resonance excitation intensity less than 0.03 W/cm2. In addition, we controllably vary the in-plane directionality of the liquid crystal molecules in the planar state by altering the linear polarization of the incident excitation. Using discrete dipole simulations and control measurements, we establish spectral selectivity in this new and interesting perspective for photonic application using low light power.
“Self-assembled nanoparticle micro-shells templated by liquid crystal sorting”A. R. Rodarte, B.H. Cao, H. Panesar, R.J. Pandolfi, M. Quint, L. Edwards, S. Ghosh, J.E. Hein and L.S. Hirst, Soft Matter, 10.1039/C4SM02326A (2015) Link
A current goal in nanotechnology focuses on the assembly of different nanoparticle types into 3D organized structures. In this paper we report the use of a liquid crystal host phase in a new process for the generation of micron-scale vesicle-like nanoparticle shells stabilized by ligand–ligand interactions. The constructs formed consist of a robust, thin spherical layer, composed of closely packed quantum dots (QDs) and stabilized by local crystallization of the mesogenic ligands. Ligand structure can be tuned to vary QD packing within the shell and made UV cross-linkable to allow for intact shell extraction into toluene. The assembly method we describe could be extended to other nanoparticle types (metallic, magnetic etc.), where hollow shell formation is controlled by thermally sorting mesogen-functionalized nanoparticles in a liquid crystalline host material at the isotropic to nematic transition. This process represents a versatile method for making non-planar 3D nano-assemblies.
Congratulations go to Ron Pandolfi, the latest PhD graduate from the Hirst group. Ron’s PhD defense was on Monday Dec 8th, where he presented his thesis on “Self-assembly and Design of Tunable Soft Materials”
During his time in the lab Ron’s work has included molecular dynamics simulations of semi-flexible polymers and x-ray characterization of different soft systems.
Ron was recently hired at the Advanced Light Source, Lawrence Berkeley National Lab in Berkeley, CA where he’ll be working with soft matter x-ray team.
Professor Hirst is the author of “Fundamentals of Soft Matter Science”
Fundamentals of Soft Matter Science introduces and explores the scientific study of soft matter and molecular self-assembly, covering the major classifications of materials, their structure and characteristics, and everyday applications.