The ability to perform high-throughput, high-purity, multi-parametric cell sorting is extremely important for many biomedical studies and clinical applications. In the past few decades, fluorescence-activated cell sorters have become the "gold standard" technique in the field. However, current cell sorters suffer from an inability to maintain cell integrity during the cell- sorting process. Conventional cell-sorting processes are reported to significantly reduce cell viability and function (30-70% reduction) for many fragile or sensitive cells such as neurons, stem cells, liver cells, dendritic cells, sperm cells, and even neutrophils from healthy individual. In addition, our recent preliminary results indicate that gene expression can be significantly altered during the cell-sorting process, even for robust cells (such as HeLa cells). These drawbacks significantly limit the usefulness of cell sorters in many biomedical studies and clinical applications and have created many unmet needs. For example, human induced pluripotent stem (iPS) cells have opened a new field for modeling human diseases using human cells directly. They can be extremely useful for drug screening and personalized medicine. However, today it is still impossible to use cell sorters or any other existing methods to isolate undifferentiated iPS cells in a high-throughput, high-purity, and high-cell-integrity manner. This unmet need has significantly hindered progress in stem cell research and therapy. Our objective is to address these unmet needs by demonstrating standing surface acoustic wave (SSAW) based, high-cell-integrity sorters. When compared to conventional sorters, the proposed SSAW cell sorter is substantially smaller and less expensive, and is expected to significantly improve post-sorting cell viability, function, and gene expression for both fragile and robust cells. In particular, we will (1) develop a SSAW-based flow cytometer that achieves sheathless, multi-color, high-throughput single-cell analysis;(2) demonstrate a high-throughput, single-cell deflecting unit using focused interdigital transducers (f-IDTs);(3) establish a fully integrated, SSAW-based cell sorter system proven with human blood samples to outperform a state-of-the-art cell sorter;and (4) demonstrate sorting of induced pluripotent stem (iPS) cells with maintained cell integrity. With unprecedented capabilities to maintain cell integrity, even for fragile cells, our proposed SSAW-based cell sorter will not only become a more compact, affordable, and easy-to-maintain replacement to the existing cell sorters, but also fill many unmet needs in both fundamental biomedical research and clinical diagnosis and therapeutics.

Public Health Relevance

The proposed project is to develop tools that can sort cells accurately while maintaining the highest cell integrity. It addresses many unmet needs and has the potential to transform both fundamental biomedical research (e.g., understanding of malaria) and clinical diagnostics and therapeutics (e.g., stem-cell-based therapies).

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
1R01GM112048-01A1
Application #
8761977
Study Section
Instrumentation and Systems Development Study Section (ISD)
Program Officer
Edmonds, Charles G
Project Start
2014-08-01
Project End
2018-07-31
Budget Start
2014-08-01
Budget End
2015-07-31
Support Year
1
Fiscal Year
2014
Total Cost
$280,091
Indirect Cost
$90,091
Name
Pennsylvania State University
Department
Engineering (All Types)
Type
Schools of Engineering
DUNS #
003403953
City
University Park
State
PA
Country
United States
Zip Code
16802
Chen, Yuchao; Wu, Mengxi; Ren, Liqiang et al. (2016) High-throughput acoustic separation of platelets from whole blood. Lab Chip 16:3466-72
Mao, Zhangming; Xie, Yuliang; Guo, Feng et al. (2016) Experimental and numerical studies on standing surface acoustic wave microfluidics. Lab Chip 16:515-24
Chen, Kejie; Wu, Mengxi; Guo, Feng et al. (2016) Rapid formation of size-controllable multicellular spheroids via 3D acoustic tweezers. Lab Chip 16:2636-43
Ahmed, Daniel; Ozcelik, Adem; Bojanala, Nagagireesh et al. (2016) Rotational manipulation of single cells and organisms using acoustic waves. Nat Commun 7:11085
Nama, Nitesh; Huang, Po-Hsun; Huang, Tony Jun et al. (2016) Investigation of micromixing by acoustically oscillated sharp-edges. Biomicrofluidics 10:024124
Xie, Yuliang; Nama, Nitesh; Li, Peng et al. (2016) Probing Cell Deformability via Acoustically Actuated Bubbles. Small 12:902-10
Guo, Feng; Mao, Zhangming; Chen, Yuchao et al. (2016) Three-dimensional manipulation of single cells using surface acoustic waves. Proc Natl Acad Sci U S A 113:1522-7
Kaynak, Murat; Ozcelik, Adem; Nama, Nitesh et al. (2016) Acoustofluidic actuation of in situ fabricated microrotors. Lab Chip 16:3532-7
Li, Peng; Mao, Zhangming; Peng, Zhangli et al. (2015) Acoustic separation of circulating tumor cells. Proc Natl Acad Sci U S A 112:4970-5
Li, Sixing; Ding, Xiaoyun; Mao, Zhangming et al. (2015) Standing surface acoustic wave (SSAW)-based cell washing. Lab Chip 15:331-8

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