3D In Vitro Neuron on a Chip for Probing Calcium Mechanostimulation

The evolution of tissue on a chip systems holds promise for mimicking the response of biological functionality of physiological systems. One important direction for tissue on a chip approaches are neuron‐based systems that could mimic neurological responses and lessen the need for in vivo experimentation. For neural research, more attention has been devoted recently to understanding mechanics due to issues in areas such as traumatic brain injury (TBI) and pain, among others. To begin to address these areas, a 3D Nerve Integrated Tissue on a Chip (NITC) approach combined with a Mechanical Excitation Testbed (MET) System is developed to impose external mechanical stimulation toward more realistic physiological environments. PC12 cells differentiated with nerve growth factor, which were cultured in a controlled 3D scaffolds, are used. The cells are labeled in a 3D NITC system with Fluo‐4‐AM to examine their calcium response under mechanical stimulation synchronized with image capture. Understanding the neural responses to mechanical stimulation beyond 2D systems is very important for neurological studies and future personalized strategies. This work will have implications in a diversity of areas including tissue‐on‐a‐chip systems, biomaterials, and neuromechanics.Developing more physiologically relevant 3D neuron integrated systems with external mechanics with synchronized image capture enables understanding of cellular responses that are important for advancing future neural studies and personalized strategies. This work examines calcium response with respect to voice coil actuator controlled force profiles.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/163437/3/adbi202000080-sup-0001-SuppMat.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/163437/2/adbi202000080_am.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/163437/1/adbi202000080.pd

Detection of Dynamic Spatiotemporal Response to Periodic Chemical Stimulation in a Xenopus Embryonic Tissue

Embryonic development is guided by a complex and integrated set of stimuli that results in collective system-wide organization that is both time and space regulated. These regulatory interactions result in the emergence of highly functional units, which are correlated to frequency-modulated stimulation profiles. We have determined the dynamic response of vertebrate embryonic tissues to highly controlled, time-varying localized chemical stimulation using a microfluidic system with feedback control. Our approach has enabled localized spatiotemporal manipulation of the steroid hormone dexamethasone (DEX) in Animal Cap (AC) tissues isolated from gastrulating Xenopus embryos. Using this approach we investigated cell-scale responses to precisely controlled stimulation by tracking the redistribution of a GFP-tagged DEX-reporter constructed from the human glucocorticoid receptor (GR). We exposed defined regions of a single AC explant to different stimulation conditions—continuous stimulation, periodic stimulation, and no stimulation. We observed collective behavior of the GR transport into the nucleus was first-order. Furthermore, the dynamic response was well-modeled by a first-order differential equation with a single time derivative. The model predicted that responses to periodic stimulations closely matched the results of the frequency-based experiments. We find that stimulation with localized bursts versus continuous stimulation can result in highly distinct responses. This finding is critical as controlled space and time exposure to growth factors is a hallmark of complex processes in embryonic development. These complex responses to cellular signaling and transport machinery were similar to emergent behaviors in other complex systems, suggesting that even within a complex embryonic tissue, the overall system can converge toward a predictive first-order response

A millimeter-wave kinetic inductance detector camera for long-range imaging through optical obscurants

Millimeter-wave imaging provides a promising option for long-range target detection through optical obscurants such as fog, which often occur in marine environments. Given this motivation, we are currently developing a 150 GHz polarization-sensitive imager using a relatively new type of superconducting pair-breaking detector, the kinetic inductance detector (KID). This imager will be paired with a 1.5 m telescope to obtain an angular resolution of 0.09° over a 3.5° field of view using 3,840 KIDs. We have fully characterized a prototype KID array, which shows excellent performance with noise strongly limited by the irreducible fluctuations from the ambient temperature background. Full-scale KID arrays are now being fabricated and characterized for a planned demonstration in a maritime environment later this year

BRIDGING MECHANICAL STIMULATION OF CELLULAR AND MOLECULAR STRUCTURE THROUGH LATTICE BASED COMPUTATIONAL SIMULATIONS

ABSTRACT Understanding the connection between mechanics and cell structure requires a critical exploration of molecular structure. One of these molecular bridges is known to be the cytoskeleton, which is involved with intracellular organization and mechanotransduction. In order to examine the structure in cells, we have developed a computational simulation that is able to probe the self-assembly of actin filaments through a lattice based Monte Carlo method. We have modeled the polymerization of these filaments based upon the interactions of globular actin through a probabilistic scheme with both inert and active proteins. The results show similar response to classic ordinary differential equations at low molecular concentrations, but a bi-phasic divergence at realistic concentrations for living mammalian cells. Further, these inert monomers have a limiting effect based upon their relative density ratios, which alter the polymerization process. Finally, by introducing localized mobility parameters, we are able to set up molecular gradients that are found in non-homogeneous protein distributions in vitro. This method and results have potential applications in cell and molecular biology as well as self assembly in inorganic systems

Mechanical loading of stem cells for improvement of transplantation outcome in a model of acute myocardial infarction: The role of loading history

Stem cell therapy for tissue repair is a rapidly evolving field and the factors that dictate the physiological responsiveness of stem cells remain under intense investigation. In this study we hypothesized that the mechanical loading history of muscle-derived stem cells (MDSCs) would significantly impact MDSC survival, host tissue angiogenesis, and myocardial function after MDSC transplantation into acutely infarcted myocardium. Mice with acute myocardial infarction by permanent left coronary artery ligation were injected with either nonstimulated (NS) or mechanically stimulated (MS) MDSCs. Mechanical stimulation consisted of stretching the cells with equibiaxial stretch with a magnitude of 10% and frequency of 0.5Hz. MS cell-transplanted hearts showed improved cardiac contractility, increased numbers of host CD31+ cells, and decreased fibrosis, in the peri-infarct region, compared to the hearts treated with NS MDSCs. MS MDSCs displayed higher vascular endothelial growth factor expression than NS cells in vitro. These findings highlight an important role for cyclic mechanical loading preconditioning of donor MDSCs in optimizing MDSC transplantation for myocardial repair. © 2012, Mary Ann Liebert, Inc

The cryomechanical design of MUSIC: a novel imaging instrument for millimeter-wave astrophysics at the Caltech Submillimeter Observatory

MUSIC (Multicolor Submillimeter kinetic Inductance Camera) is a new facility instrument for the Caltech Submillimeter Observatory (Mauna Kea, Hawaii) developed as a collaborative effect of Caltech, JPL, the University of Colorado at Boulder and UC Santa Barbara, and is due for initial commissioning in early 2011. MUSIC utilizes a new class of superconducting photon detectors known as microwave kinetic inductance detectors (MKIDs), an emergent technology that offers considerable advantages over current types of detectors for submillimeter and millimeter direct detection. MUSIC will operate a focal plane of 576 spatial pixels, where each pixel is a slot line antenna coupled to multiple detectors through on-chip, lumped-element filters, allowing simultaneously imaging in four bands at 0.86, 1.02, 1.33 and 2.00 mm. The MUSIC instrument is designed for closed-cycle operation, combining a pulse tube cooler with a two-stage Helium-3 adsorption refrigerator, providing a focal plane temperature of 0.25 K with intermediate temperature stages at approximately 50, 4 and 0.4 K for buffering heat loads and heat sinking of optical filters. Detector readout is achieved using semi-rigid coaxial cables from room temperature to the focal plane, with cryogenic HEMT amplifiers operating at 4 K. Several hundred detectors may be multiplexed in frequency space through one signal line and amplifier. This paper discusses the design of the instrument cryogenic hardware, including a number of features unique to the implementation of superconducting detectors. Predicted performance data for the instrument system will also be presented and discussed