Nanoscale control of matter is critical to the design of integrated nanosystems. Here, we describe a method to dynamically control directionality of microtubule (MT) motion using programmable magnetic fields. MTs are combined with magnetic quantum dots (i.e., MagDots) that are manipulated by external magnetic fields provided by magnetic nanowires. MT shuttles thus undergo both ATP-driven and externally-directed motion with a fluorescence component that permits simultaneous visualization of shuttle motion. This technology is used to alter the trajectory of MTs in motion and to pin MT motion. Such an approach could be used to evaluate the MT-kinesin transport system and could serve as the basis for improved lab-on-a-chip technologies based on MT transport.
Semiconductor quantum dots (QDs) have achieved initial success as biomedical imaging agents. However, significant cytotoxicity in the biological environment prohibits their use in vivo. Here, we introduce nanocomposites composed of carbon dots (C-dots) in poly(lactic-co-glycolic acid) (PLGA) carriers as possible imaging agents for in vivo applications. An initial hurdle to clinical use is overcome by synthesizing C-dots with commercially available carbon black precursors, permitting scalable nanomanufacturing. These fluorescent nanoparticles, which have a mean diameter of ~1 nm, display a disordered graphitic structure. To overcome a second clinical hurdle (i.e., rapid renal clearance of nanoparticles <~6 nm in diameter), C-dots were encapsulated in biodegradable PLGA nanospheres. The resulting nanocomposites showed a mean diameter of 344 ± 23 nm, which should reduce renal clearance. With further optimization, nanocarriers could be optimized to sizes <200 nm to reduce accumulation in the reticuloendothelial system (RES). Toxicity of both C-dots and C-dot-PLGA nanocomposites was evaluated using HepG2 liver cell lines. Unlike QDs, which can induce toxicological responses at concentrations as low as 0.005 mg/mL, C-dots exhibited cytotoxicity at concentrations greater than 0.2 mg/mL, while their derived nanocomposites did not exhibit cytotoxicity at any concentration tested (i.e., 0.02 mg/mL, 0.1 mg/mL and 0.2 mg/mL).
The microtubule (MT)-kinesin system has been extensively studied because of its role in cellular processes, as well as its potential use for controllably transporting objects at the nanoscale. Thus, there is substantial interest in methods to evaluate MT properties, including bending radius and the binding energy of kinesin motor proteins to MT tracks. Current methods to identify these properties include optical tweezers, microfluidic devices, and magnetic fields. Here, the use of magnetic quantum dots (i.e., MagDots) is evaluated as a method to study MT-kinesin interactions via applied magnetic forces. Magnetic fields are generated using a magnetic needle whose field gradient is quantified by finite element modeling (FEM). Magnetic force is applied to MagDot-labeled MTs and demonstrated sufficient to steer and detach MTs from kinesin-coated surfaces. Taking advantage of the dual-functionality of MagDots, the magnetic force experienced by a single MagDot and the number of MagDots on MTs are determined. The total force exerted on MTs by MagDots is estimated to be %0.94-2.47 pN. This approach could potentially be used to interrogate MT properties and MT-kinesin interactions, enhancing our biological understanding of this system and enabling further development of MT shuttles for nanotransport.
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