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High-throughput high-content single cell analysis by multichannel stimulatedRaman flow cytometry

Award Number: R01GM118471
ORGANIZATION: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)
PERIOD OF PERFORMANCE START DATE: 08/01/2016
PERIOD OF PERFORMANCE END DATE: 04/30/2021

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High-throughput high-content single cell analysis by multichannel stimulatedRaman flow cytometry - ¿ DESCRIPTION (provided by applicant): Flow cytometry is one of the most important tools for high-throughput single cell analysis. Fluorescent labeling acts as the primary approach for cellular analysis in flow cytometry. Nevertheless, fluorescent tags are not applicable to all cases especially small molecules (e.g. metabolites) for which labeling may significantly perturb their properties. Raman spectroscopic signals arising from inherent molecular vibrations provide a key approach to detect specific molecules inside cells and to differentiate cellular state. Raman-based microfluidic devices have been reported. However, the very small cross section of spontaneous Raman scattering results in low Raman signal level and consequently long data acquisition time, which is not compatible with the high- speed flow condition. The long-term goal of the proposed project is to establish a high-throughput high-content single cell analysis platform using molecular fingerprint vibrations as contrast. The specific objective of current application is to develop a vibrational spectroscopic cytometer based on the stimulated Raman scattering (SRS) process. Several recent advances in the Ji-Xin Cheng (PI) lab, including the highly sensitive femtosecond SRS imaging, lock-in free SRS signal detection and a tuned amplifier array for multiplex SRS imaging, pave the foundation for the planned instrumentation. The PI has assembled an interdisciplinary team for the proposed study. Dr. J. Paul Robinson (co-PI) is a leader in development and applications of fluorescence-based flow cytometer and he will bring expertise to the design of fluidics and multichannel detection systems. Dr. Bartek Rajwa (co-PI) will provide expertise for spectroscopic cytometry data analysis and machine learning. The team will design and construct a SRS flow cytometer by multichannel detection of dispersed SRS signal (Aim 1), construct a tandem system able to collect SRS and fluorescence data (Aim 2), develop spectral un-mixing and machine-learning analysis tools able to combine the information obtained from SRS spectra and labeled biomarkers for functional classification of cells (Aim 3), and validate the capability of SRS flow cytometer for label-free detection of single-cell metabolism (Aim 4). With a speed of analyzing thousands of cells per second, SRS flow cytometer will enable high-throughput analysis of single-cell chemical content which is beyond the reach by fluorescence-based flow cytometer.


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: 2021 ( Subtotal = -$1,281 )
2021	2019	TRUSTEES OF BOSTON UNIVERSITY	1 SILBER WAY	BOSTON	MA	02215	SUFFOLK	USA	Biomedical Research and Research Training	000	4	8/28/2021	NON-COMPETING CONTINUATION	-$1,281
														Subtotal = -$1,281

Issue Date FY: 2019 ( Subtotal = $388,535 )
2019	2019	TRUSTEES OF BOSTON UNIVERSITY	1 SILBER WAY	BOSTON	MA	02215	SUFFOLK	USA	Biomedical Research and Research Training	000	4	5/14/2019	NON-COMPETING CONTINUATION	$388,535
														Subtotal = $388,535

Issue Date FY: 2018 ( Subtotal = $388,379 )
2018	2018	TRUSTEES OF BOSTON UNIVERSITY	1 SILBER WAY	BOSTON	MA	02215	SUFFOLK	USA	Biomedical Research and Research Training	002	3	4/27/2018	NON-COMPETING CONTINUATION	$388,379
2018	2017	TRUSTEES OF BOSTON UNIVERSITY	1 SILBER WAY	BOSTON	MA	02215	SUFFOLK	USA	Biomedical Research and Research Training	001	2	11/11/2017	CHANGE OF GRANTEE / TRAINING INSTITUTION / AWARDING INSTITUTION	$19,996
2018	2016	PURDUE UNIVERSITY	401 SOUTH GRANT ST	WEST LAFAYETTE	IN	47907	TIPPECANOE	USA	Biomedical Research and Research Training	000	1	11/11/2017	NEW	-$19,996
														Subtotal = $388,379

Issue Date FY: 2017 ( Subtotal = $403,725 )
2017	2017	TRUSTEES OF BOSTON UNIVERSITY	1 SILBER WAY	BOSTON	MA	02215	SUFFOLK	USA	Biomedical Research and Research Training	000	2	7/7/2017	CHANGE OF GRANTEE / TRAINING INSTITUTION / AWARDING INSTITUTION	$403,725
2017	2017	TRUSTEES OF BOSTON UNIVERSITY	1 SILBER WAY	BOSTON	MA	02215	SUFFOLK	USA	Biomedical Research and Research Training	002	2	9/26/2017	CHANGE OF GRANTEE / TRAINING INSTITUTION / AWARDING INSTITUTION	$120,825
2017	2016	PURDUE UNIVERSITY	401 SOUTH GRANT ST	WEST LAFAYETTE	IN	47907	TIPPECANOE	USA	Biomedical Research and Research Training	001	1	9/26/2017	NEW	-$120,825
														Subtotal = $403,725

Issue Date FY: 2016 ( Subtotal = $428,677 )
2016	2016	PURDUE UNIVERSITY	610 PURDUE HALL	WEST LAFAYETTE	IN	47907	TIPPECANOE	USA	Biomedical Research and Research Training	000	1	7/25/2016	NEW	$428,677
														Subtotal = $428,677

Grand Total All Awards = $1,608,035

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High-throughput high-content single cell analysis by multichannel stimulatedRaman flow cytometry

Award Number: R01GM118471

ORGANIZATION: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

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

PERIOD OF PERFORMANCE START DATE: 08/01/2016

PERIOD OF PERFORMANCE END DATE: 04/30/2021

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