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An Affordable and Versatile Two-Dimensional Cell Isolation and Tracking Platform Based on Image Machine Learning and Maskless Photolithography Single Cell Encapsulation

Award Number: R21EB033102
ORGANIZATION: NATIONAL INSTITUTE Of BIOMEDICAL IMAGING & BIOENGINEERING
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)
PERIOD OF PERFORMANCE START DATE: 09/01/2022
PERIOD OF PERFORMANCE END DATE: 06/30/2026

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An Affordable and Versatile Two-Dimensional Cell Isolation and Tracking Platform Based on Image Machine Learning and Maskless Photolithography Single Cell Encapsulation - An Affordable and Versatile Two-Dimensional Cell Isolation and Tracking Platform Based on Image Machine Learning and Maskless Photolithography Single Cell Encapsulation Current commercial cell sorters typically use sheath flow to align cells into a single profile and sort cells based on fluorescence signal or images. The single profile alignment limits the throughput and requires complex hardware and expensive equipment for high-speed sorting. The usage of high-speed sheath flow also generates high stress on cells, which makes it not suitable for fragile or sensitive cells such as stem cells for downstream application. Some sticky cells such as monocytes or too many dead cells in the sample can interrupt or even clog the flow. Such cell sorter also usually requires a significant amount of starting cell number. Considering the yield, purity, and fluid dead volume, it is challenging to sort out cells of rare population such as subset of stem cells or circulating tumor cells in blood sample. There are strong needs from small labs for an affordable and versatile cell sorting platform applicable to a variety of cell types. The objectives of the proposed work are to: 1. Develop a high-speed machine learning-based cell classification module. The module will enable real-time detection of target cells inside a wide microfluidic channel based on brightfield or fluorescent images. 2. Develop a stop flow lithography-based 2D cell sorting platform in combination with acoustic field cell array patterning that will generate encoded encapsulations of target cells of different sizes. 3. Integrate the machine learning detection and maskless lithography with the size-based filtering/sorting of the cell into an affordable cell sorter. The setup can be mounted onto existing microscope and high-resolution camera, along with a web-lab flow controller and a UV projector, makes a versatile and affordable cell sorter. The proposed method can sort multiple cell types based on high content image information and machine learning. This eliminates the dependency on specific antibody types which is the basis of fluorescence-activated cell sorting (FACS) or magnetics-activated cell sorting (MACS). The proposed method can use simple microfluidic devices for sorting different types of target cells in high purity with minimum requirement on starting cell number, thus is applicable to rare subset of a large sample or rare cells. Maskless lithography based on digital micromirror device (DMD) is used to stamp encoded ID to track individual cells which is convenient for downstream analysis. The 2D wide platform can avoid high shear flow-induced cell damage or property change in the cell sorting channel, thus is suitable for gentle cells such as stem cells. The wide channel can also avoid the potential cell clogging problem in a regular cell sorter. By updating the machine learning algorithm and sharing datasets and pre-trained models, as well as the availability of cameras and projectors of better resolution, the proposed project leads to an affordable, expandable, powerful, and universal cell sorting platform.


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 = $0 )
2025	2023	LEHIGH UNIVERSITY	526 BRODHEAD AVE	BETHLEHEM	PA	18015	NORTHAMPTON	USA	Discovery and Applied Research for Technological Innovations to Improve Human Health	000	2	6/3/2025	NON-COMPETING CONTINUATION	$0
2025	2023	LEHIGH UNIVERSITY	526 BRODHEAD AVE	BETHLEHEM	PA	18015	NORTHAMPTON	USA	Discovery and Applied Research for Technological Innovations to Improve Human Health	001	2	7/23/2025	NON-COMPETING CONTINUATION	$0
														Subtotal = $0

Issue Date FY: 2023 ( Subtotal = $237,163 )
2023	2023	LEHIGH UNIVERSITY	526 BRODHEAD AVE	BETHLEHEM	PA	18015	NORTHAMPTON	USA	Discovery and Applied Research for Technological Innovations to Improve Human Health	001	2	9/20/2023	NON-COMPETING CONTINUATION	$0
2023	2023	LEHIGH UNIVERSITY	526 BRODHEAD AVE	BETHLEHEM	PA	18015	NORTHAMPTON	USA	Discovery and Applied Research for Technological Innovations to Improve Human Health	000	2	6/26/2023	NON-COMPETING CONTINUATION	$237,163
														Subtotal = $237,163

Issue Date FY: 2022 ( Subtotal = $197,023 )
2022	2022	LEHIGH UNIVERSITY	526 BRODHEAD AVE	BETHLEHEM	PA	18015	NORTHAMPTON	USA	Discovery and Applied Research for Technological Innovations to Improve Human Health	000	1	8/15/2022	NEW	$197,023
														Subtotal = $197,023

Grand Total All Awards = $434,186

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An Affordable and Versatile Two-Dimensional Cell Isolation and Tracking Platform Based on Image Machine Learning and Maskless Photolithography Single Cell Encapsulation

Award Number: R21EB033102

ORGANIZATION: NATIONAL INSTITUTE Of BIOMEDICAL IMAGING & BIOENGINEERING

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

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

PERIOD OF PERFORMANCE START DATE: 09/01/2022

PERIOD OF PERFORMANCE END DATE: 06/30/2026

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