Super-resolution imaging of subcellular structures and dynamics during non-genetic biological modulation
Abstract
Subcellular domains are highly compartmentalized and heterogeneous. The ability to modulate and sense cells at the level of organelles and extracellular vesicles is critical to future precision medicine. Biological modulation techniques such as electrical stimulation and pharmacological perturbation can simultaneously affect many adjacent cells and multiple cellular processes in addition to the target population. However, they have slow control of kinetics and weak reversibility. Therefore, the reliability and robustness of conclusions that can be obtained with such methods are limited. In this regard, it is crucial to have highly localized stimulation devices that control well-defined bioelectric activities with organelle-level selectivity, high temporal precision, and rapid reversibility. In concert, we need high accuracy imaging technologies to provide spatiotemporal readouts of the subcellular activities. In this proposal, we will request an ONI NanoImager, a suite of multi-modal super-resolution imaging techniques, and integrate it with the nano-bioelectronics stimulation platform. The new equipment would allow for the first integration of super-resolution imaging with nano-bioelectronics probing. We will use the ONI NanoImager to explore the biophysical processes that control organelle functionality and to manipulate and sense the intracellular bioelectric signaling rationally. We will achieve bioelectric modulation and imaging of intracellular organelles and extracellular vesicles. We will characterize their individual bioelectric properties and interactions among different organelles through localized stimulation and simultaneous imaging, which represents an unexplored frontier. We will also use this integrated platform to study the neuron-glia interactions with unprecedented spatial resolutions. For the nano-bioelectronics components, we will prepare two-dimensional or three-dimensional microelectrode arrays for electrochemical stimulation. We will also synthesize freestanding nanomaterials (such as silicon, silicon carbide and gold-based structures) for photothermal and photoelectrochemical modulation of cells and tissues. Next, with super-resolution imaging, we will characterize the effect of bioelectrical stimulation on the organelle or extracellular structures, levels of lumenal ions such Ca2+ and organelle membrane potential; these experiments will measure the induced bioelectric changes in organelles and their electrochemical coupling intra- and intercellularly. Finally, we will use neuron-glia co-culture system to study the effect of the non-genetic stimulation on glia-enabled neuromodulation and extracellular vesicle production. Successfully integrating both approaches - sub-cellular modulation and molecular-scale imaging - can open a wide range of scientific questions in intra- and intercellular signaling and the organization of bioelectric pathways in single cells at nanoscale spatial resolution. It can potentially provide new insights to guide the rational design of electroceuticals and yield fundamentally different and effective alternatives to the existing modulation devices.
Document Details
- Document Type
- DoD Grant Award
- Publication Date
- Jan 04, 2021
- Source ID
- W911NF2110064
Entities
People
- Bozhi Tian
Organizations
- Army Contracting Command
- United States Army
- University of Chicago