Wednesday, July 2, 2025 7/2/2025

Multiplex Imaging of Brain Activity and Plasticity with Optimized FRET/FLIM-based Sensors

Award Number: U01NS128655
ORGANIZATION: NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE
OPDIV: NIH
AWARD CLASS: COOPERATIVE AGREEMENT
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)
PERIOD OF PERFORMANCE START DATE: 01/01/2023
PERIOD OF PERFORMANCE END DATE: 12/31/2026

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Multiplex Imaging of Brain Activity and Plasticity with Optimized FRET/FLIM-based Sensors - Project Summary Plasticity is a fundamental aspect of neuronal circuits across all species. It is at the base of learning and memory, sensory adaption, and many disease-related processes such as addiction, chronic pain or regeneration. On the molecular level biochemical mechanisms have been well described, but little is known on how these are coordinated in space and time within neuronal circuits of living brains. To elucidate the circuit operation of plasticity in vivo we here propose to develop and validate highly sensitive, red FRET/FLIM sensors for simultaneous use with green calcium sensors, allowing imaging of neuronal calcium activity and biochemical signaling dynamics in individual synapses and neuronal populations. We will focus on plasticity-linked biochemical events involved in synaptic plasticity and spine morphogenesis. FRET/FLIM sensors will allow for the accurate measurement of small changes, even in the presence of brain movement during behavior. In aim 1, we will develop highly sensitive red-emitting FRET/FLIM sensors using mammalian cell based protein libraries and structurally-guided large scale screening. Aim 2 will involve validation of sensors ex vivo and in vivo. Finally, sensors will be validated for use in studies on synaptic and neuronal plasticity during spatial navigation tasks in awake mice. We plan for iterative cycles of improvements in which input from biophysical analysis of sensors, validation in slices and in vivo and feed-back from early roll-out users with different animal models will be used to create successive sensor generations with ever increasing performance. Combined in vivo 2P FRET/FLIM of plasticity and 2P fluorescence imaging of calcium activity will provide a powerful new approach to study synaptic and neuronal plasticity in living organisms.


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: 2025 ( Subtotal = $958,817 )
2025	2025	NORTHWESTERN UNIVERSITY	633 CLARK ST	EVANSTON	IL	60208	COOK	USA	Extramural Research Programs in the Neurosciences and Neurological Disorders	000	3	1/28/2025	NON-COMPETING CONTINUATION	$958,817
														Subtotal = $958,817

Issue Date FY: 2024 ( Subtotal = $950,637 )
2024	2024	NORTHWESTERN UNIVERSITY	633 CLARK STREET	EVANSTON	IL	60208	COOK	USA	Extramural Research Programs in the Neurosciences and Neurological Disorders	000	2	12/29/2023	NON-COMPETING CONTINUATION	$950,637
2024	2024	NORTHWESTERN UNIVERSITY	633 CLARK ST	EVANSTON	IL	60208	COOK	USA	Extramural Research Programs in the Neurosciences and Neurological Disorders	001	2	7/12/2024	NON-COMPETING CONTINUATION	$0
														Subtotal = $950,637

Issue Date FY: 2023 ( Subtotal = $1,159,614 )
2023	2023	NORTHWESTERN UNIVERSITY	633 CLARK STREET	EVANSTON	IL	60208	COOK	USA	Extramural Research Programs in the Neurosciences and Neurological Disorders	001	1	6/12/2023	NEW	$0
2023	2023	NORTHWESTERN UNIVERSITY	633 CLARK	EVANSTON	IL	60208	COOK	USA	Extramural Research Programs in the Neurosciences and Neurological Disorders	000	1	12/28/2022	NEW	$1,159,614
														Subtotal = $1,159,614

Grand Total All Awards = $3,069,068

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Multiplex Imaging of Brain Activity and Plasticity with Optimized FRET/FLIM-based Sensors

Award Number: U01NS128655

ORGANIZATION: NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE

OPDIV: NIH

AWARD CLASS: COOPERATIVE AGREEMENT

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

PERIOD OF PERFORMANCE START DATE: 01/01/2023

PERIOD OF PERFORMANCE END DATE: 12/31/2026

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