Microfluidic single cell analysis: from promise to practice

Lecault, Véronique; White, Alexander D.; Singhal, Anupam; Hansen, Carl L.

doi:10.1016/j.cbpa.2012.03.022

Cited by 215 publications

(163 citation statements)

References 81 publications

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“…''Labon-a-chip'' devices and systems have become a conspicuous feature of cell analyses due to small reagent volumes, dynamic control of reagents, high throughput, biocompatibility, and sensitivity. 55,56 Microfluidic analysis methods have long been a focus of technology development; it is only recently that these tools have impacted stem cell research. The maturation of microfluidics from its infancy into adolescence over the past 10 years has yielded many approaches to trap cells and manipulate their microenvironments in defined ways.…”

Section: Current Stem Cell Analysis Methods and Their Limitationsmentioning

confidence: 99%

“…The maturation of microfluidics from its infancy into adolescence over the past 10 years has yielded many approaches to trap cells and manipulate their microenvironments in defined ways. [55][56][57][58][59] The simplest strategy for isolating cells involves the placement of weirs or traps that lie in line with the fluid flow. [60][61][62] The pressure exerted by the flowing fluids holds the cells in place for imaging cytometry and the measurement of some dynamic properties, including calcium flux on stimulation, 63 cell motility, 64 or proliferation.…”

Section: Current Stem Cell Analysis Methods and Their Limitationsmentioning

confidence: 99%

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Technology Advancement for Integrative Stem Cell Analyses

Jeong

Choi

Lee

2014

Tissue Engineering Part B: Reviews

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Scientists have endeavored to use stem cells for a variety of applications ranging from basic science research to translational medicine. Population-based characterization of such stem cells, while providing an important foundation to further development, often disregard the heterogeneity inherent among individual constituents within a given population. The population-based analysis and characterization of stem cells and the problems associated with such a blanket approach only underscore the need for the development of new analytical technology. In this article, we review current stem cell analytical technologies, along with the advantages and disadvantages of each, followed by applications of these technologies in the field of stem cells. Furthermore, while recent advances in micro/nano technology have led to a growth in the stem cell analytical field, underlying architectural concepts allow only for a vertical analytical approach, in which different desirable parameters are obtained from multiple individual experiments and there are many technical challenges that limit vertically integrated analytical tools. Therefore, we propose-by introducing a concept of vertical and horizontal approach-that there is the need of adequate methods to the integration of information, such that multiple descriptive parameters from a stem cell can be obtained from a single experiment.

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Section: Current Stem Cell Analysis Methods and Their Limitationsmentioning

confidence: 99%

Section: Current Stem Cell Analysis Methods and Their Limitationsmentioning

confidence: 99%

Technology Advancement for Integrative Stem Cell Analyses

Jeong

Choi

Lee

2014

Tissue Engineering Part B: Reviews

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“…mLSI is particularly well suited for single cell analysis and manipulation; several excellent review articles have already summarized recent studies [50][51][52], which generally focus on single cell imaging [24 ,26,31,32 ,53-55] or single cell genomic/ proteomic analysis [19,30,56 ,57 ,58,59]. Another field of application of mLSI is protein synthesis and characterization.…”

Section: Biological and Chemical Applicationsmentioning

confidence: 99%

Recent developments in microfluidic large scale integration

Araci

Brisk

2014

Current Opinion in Biotechnology

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In 2002, Thorsen et al. integrated thousands of micromechanical valves on a single microfluidic chip and demonstrated that the control of the fluidic networks can be simplified through multiplexors [1]. This enabled realization of highly parallel and automated fluidic processes with substantial sample economy advantage. Moreover, the fabrication of these devices by multilayer soft lithography was easy and reliable hence contributed to the power of the technology; microfluidic large scale integration (mLSI). Since then, mLSI has found use in wide variety of applications in biology and chemistry. In the meantime, efforts to improve the technology have been ongoing. These efforts mostly focus on; novel materials, components, micromechanical valve actuation methods, and chip architectures for mLSI. In this review, these technological advances are discussed and, recent examples of the mLSI applications are summarized. Recent advancements in microfluidic technologies have created new opportunities in the fields of biology and chemistry through miniaturization and automation of common processes, including mixing, metering, trapping, reaction, filtering, and separation, among others. Microfluidic large scale integration (mLSI) enables the fabrication of microfluidic chips containing hundreds or more of these functions using well-established lithography techniques, similar in principle to electronic LSI in the semiconductor industry [1]. Small feature dimensions, abundant spatial parallelism, and control over fluid flow provide mLSI technology with advantages such as high throughput, precise metering, automation and low reagent consumption. The implications of mLSI technology can especially be seen in recent advances in single cell, genomic, and protein analysis.

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“…[12][13][14][15] Many microfluidic devices for single-cell gene expression analysis have been demonstrated including single cell isolation, lysis, complementary DNA (cDNA) synthesis, 3,16 and real-time quantitative polymerase chain reaction (qPCR) 17 following off-chip protocol. 18,19 Especially, digital polymerase chain reaction (PCR) devices have supplied an efficient and precise platform for single-cell analysis because of their well-fit scale and higher precision. [20][21][22][23] Digital PCR chip with microvalves have been used to map the cellular subpopulations in normal tissues and tumors.…”

Section: Introductionmentioning

confidence: 99%

Single cell digital polymerase chain reaction on self-priming compartmentalization chip

Zhu

Qiu

et al. 2017

Biomicrofluidics

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Single cell analysis provides a new framework for understanding biology and disease, however, an absolute quantification of single cell gene expression still faces many challenges. Microfluidic digital polymerase chain reaction (PCR) provides a unique method to absolutely quantify the single cell gene expression, but only limited devices are developed to analyze a single cell with detection variation. This paper describes a self-priming compartmentalization (SPC) microfluidic digital polymerase chain reaction chip being capable of performing single molecule amplification from single cell. The chip can be used to detect four single cells simultaneously with 85% of sample digitization. With the optimized protocol for the SPC chip, we first tested the ability, precision, and sensitivity of our SPC digital PCR chip by assessing β-actin DNA gene expression in 1, 10, 100, and 1000 cells. And the reproducibility of the SPC chip is evaluated by testing 18S rRNA of single cells with 1.6%–4.6% of coefficient of variation. At last, by detecting the lung cancer related genes, PLAU gene expression of A549 cells at the single cell level, the single cell heterogeneity was demonstrated. So, with the power-free, valve-free SPC chip, the gene copy number of single cells can be quantified absolutely with higher sensitivity, reduced labor time, and reagent. We expect that this chip will enable new studies for biology and disease.

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Microfluidic single cell analysis: from promise to practice

Cited by 215 publications

References 81 publications

Technology Advancement for Integrative Stem Cell Analyses

Technology Advancement for Integrative Stem Cell Analyses

Recent developments in microfluidic large scale integration

Single cell digital polymerase chain reaction on self-priming compartmentalization chip

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