Virus Reduction Neutralization Test: A Single-Cell Imaging High-Throughput Virus Neutralization Assay for Dengue

ABSTRACTCancer stem cells (CSCs) are a heterogeneous and dynamic population that stands at the top of tumor cellular hierarchy and is responsible for maintenance of the tumor microenvironment. As methods of CSC isolation and functional interrogation advance, there is a need for a reliable and accessible quantitative approach to assess heterogeneity and state transition dynamics in CSCs. We developed a High-throughput Automated Single Cell Imaging Analysis (HASCIA) approach for quantitative assessment of protein expression with single cell resolution and applied the method to investigate spatiotemporal factors that influence CSC state transition using glioblastoma (GBM) CSC as a model system. We were able to validate the quantitative nature of this approach through comparison of the protein expression levels determined by HASCIA to those determined by immunoblotting. A virtue of HASCIA was exemplified by detection of a subpopulation of SOX2-low cells, which expanded in fraction size during state transition. HASCIA also revealed that CSCs were committed to loose stem cell state at an earlier time point than the average SOX2 level decreased. Functional assessment of stem cell frequency in combination with quantification of SOX2 expression by HASCIA defined a stable cut-off of SOX2 expression level for stem cell state. We also developed an approach to assess local cell density and found that denser monolayer areas possess higher average levels of SOX2, higher cell diversity and a presence of a sub-population of slowly proliferating SOX2-low CSCs. HASCIA is an open source software that facilitates understanding the dynamics of heterogeneous cell population such as that of CSCs and their progeny. It is a powerful and easy-to-use image analysis and statistical analysis tool available athttps://hascia.lerner.ccf.org.

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Single Cell Imaging Cytometry-Based High-Throughput Analysis of Drug-Induced Cardiotoxicity

High-Throughput Screening Methods in Toxicity Testing ◽

10.1002/9781118538203.ch21 ◽

2013 ◽

pp. 385-402

Author(s):

Min Jung Kim ◽

Joon Myong Song

Keyword(s):

Single Cell ◽

High Throughput ◽

Cell Imaging ◽

Drug Induced ◽

High Throughput Analysis ◽

Throughput Analysis ◽

Single Cell Imaging ◽

Imaging Cytometry

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Dynamic analysis of MAPK signaling using a high-throughput microfluidic single-cell imaging platform

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.0813416106 ◽

2009 ◽

Vol 106 (10) ◽

pp. 3758-3763 ◽

Cited By ~ 114

Author(s):

R. J. Taylor ◽

D. Falconnet ◽

A. Niemisto ◽

S. A. Ramsey ◽

S. Prinz ◽

...

Keyword(s):

Dynamic Analysis ◽

Single Cell ◽

High Throughput ◽

Cell Imaging ◽

Mapk Signaling ◽

Single Cell Imaging

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Review: imaging technologies for flow cytometry

Lab on a Chip ◽

10.1039/c6lc01063f ◽

2016 ◽

Vol 16 (24) ◽

pp. 4639-4647 ◽

Cited By ~ 87

Author(s):

Yuanyuan Han ◽

Yi Gu ◽

Alex Ce Zhang ◽

Yu-Hwa Lo

Keyword(s):

Flow Cytometry ◽

Single Cell ◽

High Throughput ◽

Cell Imaging ◽

Imaging Technologies ◽

Imaging Flow Cytometry ◽

Single Cell Imaging

Imaging flow cytometry combines the single-cell imaging capabilities of microscopy with the high-throughput capabilities of conventional flow cytometry. This article describes recent imaging flow cytometry technologies and their challenges.

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A Novel High-Throughput Vaccinia Virus Neutralization Assay and Preexisting Immunity in Populations from Different Geographic Regions in China

PLoS ONE ◽

10.1371/journal.pone.0033392 ◽

2012 ◽

Vol 7 (3) ◽

pp. e33392 ◽

Cited By ~ 16

Author(s):

Qiang Liu ◽

Weijin Huang ◽

Jianhui Nie ◽

Rong Zhu ◽

Dongying Gao ◽

...

Keyword(s):

Vaccinia Virus ◽

High Throughput ◽

Neutralization Assay ◽

Virus Neutralization ◽

Virus Neutralization Assay ◽

Geographic Regions

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From imaging a single cell to implementing precision medicine: an exciting new era

Emerging Topics in Life Sciences ◽

10.1042/etls20210219 ◽

2021 ◽

Author(s):

Loukia G. Karacosta

Keyword(s):

Precision Medicine ◽

Single Cell ◽

High Throughput ◽

Cell Biology ◽

Cell Imaging ◽

Machine Learning Algorithms ◽

Biomedical Sciences ◽

Computational Tools ◽

New Era ◽

Single Cell Imaging

In the age of high-throughput, single-cell biology, single-cell imaging has evolved not only in terms of technological advancements but also in its translational applications. The synchronous advancements of imaging and computational biology have produced opportunities of merging the two, providing the scientific community with tools towards observing, understanding, and predicting cellular and tissue phenotypes and behaviors. Furthermore, multiplexed single-cell imaging and machine learning algorithms now enable patient stratification and predictive diagnostics of clinical specimens. Here, we provide an overall summary of the advances in single-cell imaging, with a focus on high-throughput microscopy phenomics and multiplexed proteomic spatial imaging platforms. We also review various computational tools that have been developed in recent years for image processing and downstream applications used in biomedical sciences. Finally, we discuss how harnessing systems biology approaches and data integration across disciplines can further strengthen the exciting applications and future implementation of single-cell imaging on precision medicine.

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