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Automated Isolation of Single Cells Using High-Resolution and Subcellular Imaging

Award Number: R44GM119888
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: 09/01/2013
PERIOD OF PERFORMANCE END DATE: 02/29/2020

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Automated Isolation of Single Cells Using High-Resolution and Subcellular Imaging - PROJECT SUMMARY Pairing detailed phenotypic characterization of cells with downstream molecular analysis represents a significant challenge, especially at the level of single cells. Cell Microsystems has developed and commercialized the CellRaft™ System, which is currently the only commercially available means of imaging cells using brightfield and/or multi-‐‑channel fluorescence prior to single cell isolation and downstream molecular analysis. Investigators interested in pairing phenotypic data with genomic analysis at the single cell level have employed the CellRaft System for a range of innovative studies including stem cell differentiation, tumor cell physiology and screening of neurons. However, the materials used in the currently commercially available CellRaft Arrays are not optimized for imaging quality and were selected for other properties such as the re-‐‑sealing behavior of the elastomer and cell culture compatibility of the plastic. During the Phase I program, alternative materials for the CellRaft Array were explored for improved optical properties such as refractive index agreement and reduced autofluorescence. One particular alternative materials combination exhibited favorable performance in both criteria leading to dramatically improved cell imaging performance. Here, we refer to this version of the product as the High Resolution, or HR-‐‑CellRaft. The focus of our Phase II proposal is to commercialize the HR-‐‑CellRaft for users interested in pairing detailed phenotypic imaging not possible on the current CellRaft Array (e.g. subcellular protein localization, organelle physiology, and semi-‐‑quantitative fluorescent signals), as well as automate the CellRaft System workflow. In a parallel program, we are developing a highly automated instrument, the Automated Isolation and Retrieval (AIR™) System for single cell imaging and isolation employing the current CellRaft Array. During Phase II we will implement several design revisions to the optical capabilities of the AIR System as well as its software-‐‑based image analysis algorithms to better leverage the HR-‐‑CellRaft’s improved optical properties. The new proposed instrument is referred to here as the CLEAR AIR™ System. Additional fluorescence detection channels (six instead of three in the AIR™ System), higher magnification objectives (10X, 20X and 40X as opposed to the single objective -‐‑ 4X or 10X -‐‑ found on the AIR™ System) and a higher resolution camera will enhance the imaging capabilities of the system. For external validation of HR-‐‑CellRaft Array and CLEAR AIR System performance, we have identified two investigators from our Early Adopter Program with research needs uniquely satisfied by the HR-‐‑CellRaft and CLEAR AIR System. One will isolate single cells based on subcellular localization of a reporter protein and the second will isolate cells with varying levels of transgene expression, with both groups coupling these phenotypic observations to downstream single cell molecular analysis. The HR-‐‑CellRaft and the improved optical subsystem in the CLEAR AIR System meets an urgently unmet need for pairing high-‐‑resolution phenotypic imaging with downstream molecular analysis of single cells.


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: 2020 ( Subtotal = $0 )
2020	2018	CELL MICROSYSTEMS, INC.	801 CAPITOLA DR STE 10	DURHAM	NC	27713	DURHAM	USA	Biomedical Research and Research Training	000	3	7/6/2020	NON-COMPETING CONTINUATION	$0
														Subtotal = $0

Issue Date FY: 2018 ( Subtotal = $768,019 )
2018	2018	CELL MICROSYSTEMS, INC.	2 DAVIS DRIVE ROOM 119 AND 118	RESEARCH TRIANGLE PARK	NC	27709	DURHAM	USA	Biomedical Research and Research Training	000	3	3/13/2018	NON-COMPETING CONTINUATION	$768,019
														Subtotal = $768,019

Issue Date FY: 2017 ( Subtotal = $818,501 )
2017	2017	CELL MICROSYSTEMS, INC.	2 DAVIS DRIVE ROOM 119 AND 118	RESEARCH TRIANGLE PARK	NC	27709	DURHAM	USA	Biomedical Research and Research Training	000	2	2/28/2017	COMPETING CONTINUATION	$818,501
														Subtotal = $818,501

Grand Total All Awards = $1,586,520

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Automated Isolation of Single Cells Using High-Resolution and Subcellular Imaging

Award Number: R44GM119888

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: 09/01/2013

PERIOD OF PERFORMANCE END DATE: 02/29/2020

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