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SERIAL BLOCKFACE SEM LABELS FOR ASSESSING NERVOUS SYSTEM PLASTICITY

Award Number: R43MH086994
ORGANIZATION: NATIONAL INSTITUTE OF MENTAL HEALTH
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)
PERIOD OF PERFORMANCE START DATE: 09/18/2009
PERIOD OF PERFORMANCE END DATE: 10/31/2012

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SERIAL BLOCKFACE SEM LABELS FOR ASSESSING NERVOUS SYSTEM PLASTICITY - DESCRIPTION (provided by applicant): Serial blockface scanning electron microscopy (SBFSEM) is a recently developed electron microscopy serial- reconstruction technique which automates the process of sectioning and imaging blocks of tissue by incorporating a microtome into the vacuum chamber of a scanning electron microscope (SEM). SBFSEM provides resolution that is sufficient to trace even the thinnest axons and to identify synapses. It also allows the user to automatically obtain several hundred sections (50-70 nm thick) needed to completely reconstruct the connectivity of neuronal circuits, while potentially providing nanometer-scale resolution for immunogold labeling. SBFSEM will be combined with pre-embedding immunogold labeling to develop a method for the three-dimensional mapping of targets at the molecular level in very large specimens. Conducted in parallel with neuronal mapping, this will allow correlation of localization of specific molecules of interest in the context of high-resolution mapping of neuronal interconnectivity. Reagents will be developed for multiple labeling using different sized gold particles optimized for penetration, minimal background, high solubility, and maximum compatibility with specimen characteristics. Our goal is to make immunogold-SBRSEM a routine method that can be used to map targets within spatially large specimens. Three particle sizes in the range 0.8 to 5 nm will be identified that can be visualized, identified and differentiated using SBFSEM in multiple labeling studies, and used to demonstrate concomitant SBFSEM neuronal tract tracing and pre-embedding labeling of phos- phorylated and non- phosphorylated connexin 35 (Cx35) containing mixed synapses: (a) single and (b) double immunolabeling of neuronal and synaptic targets will be evaluated in the Western Mosquitofish, Gambusia affinis, using the new probes, and best protocols established for labeling and microscopy. PUBLIC HEALTH RELEVANCE: This research will provide new tools with which cell and structural biologists can correlate the observation of structures and processes in large specimens from biological systems, particularly the nervous system but also many other applications in cell biology, at the cellular and macromolecular level. These will enable new insights into how such processes work, and how molecular and macroscopic structure and function are related in normal and disease processes. This knowledge will lead to improved diagnostics and therapeutics in addition to providing improved immunogold probes and contributing to the development of nanobiotechnology.


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: 2012 ( Subtotal = $0 )
2012	2010	NANOPROBES, INC	95 HORSEBLOCK RD	YAPHANK	NY	11980	SUFFOLK	USA	Mental Health Research Grants	000	2	8/23/2012	NON-COMPETING CONTINUATION	$0
														Subtotal = $0

Issue Date FY: 2010 ( Subtotal = $361,529 )
2010	2010	NANOPROBES, INC	95 HORSEBLOCK RD	YAPHANK	NY	11980	SUFFOLK	USA	Mental Health Research Grants	001	2	7/26/2010	NON-COMPETING CONTINUATION	$361,529
2010	2009	NANOPROBES, INC	95 HORSEBLOCK RD	YAPHANK	NY	11980	SUFFOLK	USA	Mental Health Research Grants	000	1	10/16/2009	NEW	$0
														Subtotal = $361,529

Issue Date FY: 2009 ( Subtotal = $338,415 )
2009	2009	NANOPROBES, INC	95 HORSEBLOCK RD	YAPHANK	NY	11980	SUFFOLK	USA	Mental Health Research Grants	000	1	9/16/2009	NEW	$338,415
2009	2009	NANOPROBES, INC	95 HORSEBLOCK RD	YAPHANK	NY	11980	SUFFOLK	USA	Mental Health Research Grants	001	1	10/19/2009	NEW	$0
														Subtotal = $338,415

Grand Total All Awards = $699,944

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SERIAL BLOCKFACE SEM LABELS FOR ASSESSING NERVOUS SYSTEM PLASTICITY

Award Number: R43MH086994

ORGANIZATION: NATIONAL INSTITUTE OF MENTAL HEALTH

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

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

PERIOD OF PERFORMANCE START DATE: 09/18/2009

PERIOD OF PERFORMANCE END DATE: 10/31/2012

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