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Multi-angle Total Internal Reflection Structured Illumination Microscopy to Investigate Exo-Endocytosis Coupling in Living Cells

Award Number: R21NS143024
ORGANIZATION: NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)
PERIOD OF PERFORMANCE START DATE: 05/01/2026
PERIOD OF PERFORMANCE END DATE: 04/30/2028

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Multi-angle Total Internal Reflection Structured Illumination Microscopy to Investigate Exo-Endocytosis Coupling in Living Cells - PROJECT SUMMARY Neuronal communication depends on the precise coordination of synaptic vesicle exocytosis and endocytosis at presynaptic terminals. After neurotransmitter release, efficient retrieval of synaptic vesicle membranes through endocytosis is essential for sustaining neurotransmission. Despite its fundamental importance, the mechanisms linking synaptic vesicle exocytosis and endocytosis remain poorly understood, largely due to the limitations of current imaging technologies. Existing methods lack the spatial and temporal resolution needed to directly visualize these nanoscale processes in living cells. To address this critical gap, this Joint NINDS/NIMH Exploratory Neuroscience Research Grant (PA-25-150) proposal aims to develop a three-dimensional (3D) total internal reflection fluorescence structured illumination microscope (TIRF-SIM). By integrating super-resolution imaging with evanescent field nanometry, this technology will enable unprecedented visualization of exo-endocytic coupling at the nanoscale in living neurons. Our research plan consists of two key objectives: (i) We will iteratively optimize the experimental parameters of 3D TIRF-SIM using well-characterized in vitro and in vivo imaging conditions to maximize spatial and temporal resolution. (ii) Using these optimized parameters, we will evaluate the performance of 3D TIRF-SIM for high-resolution reconstruction of clathrin-coated structures both live cultured cells and tissues of Drosophila melanogaster embryos addressing long-standing questions in endocytosis research. Beyond its technological advancements, this proposal directly addresses a critical need in neuroscience research. By enabling direct visualization of synaptic vesicle dynamics at unprecedented resolution, this innovation will fundamentally advance our understanding of neuronal communication. Moreover, this breakthrough in imaging technology has the potential to catalyze the development of novel therapeutic strategies for neurological disorders associated with synaptic dysfunction.


Issue Date FY	Funding FY	Legal Entity Name	Legal Entity Address	Legal Entity City	Legal Entity State	Legal Entity Zip Code	Legal Entity COUNTY	Legal Entity COUNTRY	Assistance Listing	Award Code	Budget Year	Action Date	Action Type	Action Amount

Issue Date FY: 2026 ( Subtotal = $433,125 )
2026	2026	OHIO STATE UNIVERSITY, THE	1960 KENNY RD	COLUMBUS	OH	43210	FRANKLIN	USA	Extramural Research Programs in the Neurosciences and Neurological Disorders	000	1	4/27/2026	NEW	$433,125
														Subtotal = $433,125

Grand Total All Awards = $433,125

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Multi-angle Total Internal Reflection Structured Illumination Microscopy to Investigate Exo-Endocytosis Coupling in Living Cells

Award Number: R21NS143024

ORGANIZATION: NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)

PERIOD OF PERFORMANCE START DATE: 05/01/2026

PERIOD OF PERFORMANCE END DATE: 04/30/2028

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