30 research outputs found
Cerebellar tDCS: A Novel Approach to Augment Language Treatment Post-stroke
People with post-stroke aphasia may have some degree of chronic deficit for which current rehabilitative treatments are variably effective. Accumulating evidence suggests that transcranial direct current stimulation (tDCS) may be useful for enhancing the effects of behavioral aphasia treatment. However, it remains unclear which brain regions should be stimulated to optimize effects on language recovery. Here, we report on the therapeutic potential of right cerebellar tDCS in augmenting language recovery in SMY, who sustained bilateral MCA infarct resulting in aphasia and anarthria. We investigated the effects of 15 sessions of anodal cerebellar tDCS coupled with spelling therapy using a randomized, double-blind, sham controlled within-subject crossover trial. We also investigated changes in functional connectivity using resting state functional magnetic resonance imaging before and 2 months post-treatment. Both anodal and sham treatments resulted in improved spelling to dictation for trained and untrained words immediately after and 2 months post-treatment. However, there was greater improvement with tDCS than with sham, especially for untrained words. Further, generalization to written picture naming was only noted during tDCS but not with sham. The resting state functional connectivity data indicate that improvement in spelling was accompanied by an increase in cerebro-cerebellar network connectivity. These results highlight the therapeutic potential of right cerebellar tDCS to augment spelling therapy in an individual with large bilateral chronic strokes
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Microscale tools for improved analytical sensitivity and throughput in single-cell immunoblotting
Proteins drive nearly all cellular processes, and direct quantitation of protein abundance from single-cells is essential to understanding heterogeneous cell states.1,2 Immunoassays are widely accepted tools for performing single-cell protein detection,3,4 but protein detection by immunoaffinity alone is insufficient for precision protein characterization, as proteins with similar bindingkineticscanhavedifferentbiologicalimpacts,includingindisease.5,6,7 Toprovideacross- validation tool for protein characterization, electrophoretic cytometry immunoassays have been developed to characterize proteins by both immunoaffinity and molecular-mass through electrophoresis.8,9 Central to these assays performance is a multifunctional gel matrix that acts as a protein sieving matrix during electrophoresis and a protein scaffolding matrix during in-gel immunoblotting. In-gel immunoblotting of target proteins is widely accomplished by diffusively-driven immunoprobing, yet, this detection strategy suffers from reduced probe access to in-gel immobilized proteins via size-exclusion partitioning. Specifically, reduced probe delivery to the gel matrix in which target proteins are immobilized both (i) adversely impacts equilibrium immunocomplex formation and thus protein detection sensitivity and (ii) extends overall assay run time.In this dissertation, to improve the analytical detection capabilities and improve assay throughput in electrophoretic cytometry assays, we present methods to enhance immunoprobe delivery to hydrogel matrices, we introduce an assay design to improve throughput in single-cell immunoblotting, and we investigate reengineered sample handling designs for reduced protein losses before immobilization.
Overall, we apply fundamentals in materials science, transport and reaction phenomena, and engineering design principles for the advancement of targeted protein detection assays. We see these advancements as contributing to the broader goal of improving our understanding of cell- state in healthy and disease conditions
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3D projection electrophoresis for single-cell immunoblotting.
Immunoassays and mass spectrometry are powerful single-cell protein analysis tools; however, interfacing and throughput bottlenecks remain. Here, we introduce three-dimensional single-cell immunoblots to detect both cytosolic and nuclear proteins. The 3D microfluidic device is a photoactive polyacrylamide gel with a microwell array-patterned face (xy) for cell isolation and lysis. Single-cell lysate in each microwell is "electrophoretically projected" into the 3rd dimension (z-axis), separated by size, and photo-captured in the gel for immunoprobing and confocal/light-sheet imaging. Design and analysis are informed by the physics of 3D diffusion. Electrophoresis throughput is > 2.5 cells/s (70× faster than published serial sampling), with 25 immunoblots/mm2 device area (>10× increase over previous immunoblots). The 3D microdevice design synchronizes analyses of hundreds of cells, compared to status quo serial analyses that impart hours-long delay between the first and last cells. Here, we introduce projection electrophoresis to augment the heavily genomic and transcriptomic single-cell atlases with protein-level profiling
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3D projection electrophoresis for single-cell immunoblotting.
Immunoassays and mass spectrometry are powerful single-cell protein analysis tools; however, interfacing and throughput bottlenecks remain. Here, we introduce three-dimensional single-cell immunoblots to detect both cytosolic and nuclear proteins. The 3D microfluidic device is a photoactive polyacrylamide gel with a microwell array-patterned face (xy) for cell isolation and lysis. Single-cell lysate in each microwell is "electrophoretically projected" into the 3rd dimension (z-axis), separated by size, and photo-captured in the gel for immunoprobing and confocal/light-sheet imaging. Design and analysis are informed by the physics of 3D diffusion. Electrophoresis throughput is > 2.5 cells/s (70× faster than published serial sampling), with 25 immunoblots/mm2 device area (>10× increase over previous immunoblots). The 3D microdevice design synchronizes analyses of hundreds of cells, compared to status quo serial analyses that impart hours-long delay between the first and last cells. Here, we introduce projection electrophoresis to augment the heavily genomic and transcriptomic single-cell atlases with protein-level profiling