Cellular Analysis Using Microfluidics

Sibbitts, Jay; Sellens, Kathleen A.; Jia, Shu; Klasner, Scott A.; Culbertson, Christopher T.

doi:10.1021/acs.analchem.7b04519

Cited by 51 publications

(17 citation statements)

References 130 publications

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“…Since the development of the PDMS‐based soft lithography technique two decades ago , microfluidic devices have been extensively used to focus , trap , concentrate , and separate particles (varying from nano to micro, biological to synthetic, rigid to soft, etc.) for many biomedical, chemical, and environmental applications . Compared to their macroscopic counterparts, microfluidic devices have the following advantages: precise fluid control because of the small Reynolds number flow; low sample consumption because of the small volume of microchannels; accurate particle manipulation because of the strong confinement effect (as a result of the comparable particle and channel sizes), and as well the small Reynolds number, and so on.…”

Section: Introductionmentioning

confidence: 99%

Recent advances in direct current electrokinetic manipulation of particles for microfluidic applications

Xuan

2019

Electrophoresis

106

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Microfluidic devices have been extensively used to achieve precise transport and placement of a variety of particles for numerous applications. A range of force fields have thus far been demonstrated to control the motion of particles in microchannels. Among them, electric field‐driven particle manipulation may be the most popular and versatile technique because of its general applicability and adaptability as well as the ease of operation and integration into lab‐on‐a‐chip systems. This article is aimed to review the recent advances in direct current (DC) (and as well DC‐biased alternating current) electrokinetic manipulation of particles for microfluidic applications. The electric voltages are applied through electrodes that are positioned into the distant channel‐end reservoirs for a concurrent transport of the suspending fluid and manipulation of the suspended particles. The focus of this review is upon the cross‐stream nonlinear electrokinetic motions of particles in the linear electroosmotic flow of fluids, which enable the diverse control of particle transport in microchannels via the wall‐induced electrical lift and/or the insulating structure‐induced dielectrophoretic force.

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Section: Introductionmentioning

confidence: 99%

Recent advances in direct current electrokinetic manipulation of particles for microfluidic applications

Xuan

2019

Electrophoresis

106

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“…Compared to pavement and basal cells, the trichomes of Arabidopsis thaliana contained higher levels of metabolites from the kaempferol glycoside biosynthesis pathway ( Zhang et al, 2014 ). Selectively analyzing specialized single plant cells, e.g., parenchyma cells, guard cells, trichomes, and excretory idioblasts ( Foster, 1956 ; Valverde et al, 2001 ; Labhsetwar et al, 2014 ; Misra et al, 2015 ; Sibbitts et al, 2018 ) can provide insight into the biochemical processes and regulatory networks associated with their function.…”

Section: Introductionmentioning

confidence: 99%

“…Most biological experiments rely on bulk analysis that report on large cell population averages (millions of cells) and mask the presence of subpopulations, rare cells, and individual cellular variations. Measuring the distribution and the magnitude of biological noise is increasingly feasible through single cell analysis techniques, including fluorescence ( Paige et al, 2012 ), microfluidics ( Sibbitts et al, 2018 ), and Raman scattering ( Kuku et al, 2017 ). The limited size and volume of single cells, and the low copy numbers of certain analytes make detection, identification, and quantitation challenging.…”

Section: Introductionmentioning

confidence: 99%

Metabolic Noise and Distinct Subpopulations Observed by Single Cell LAESI Mass Spectrometry of Plant Cells in situ

et al. 2018

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Phenotypic variations and stochastic expression of transcripts, proteins, and metabolites in biological tissues lead to cellular heterogeneity. As a result, distinct cellular subpopulations emerge. They are characterized by different metabolite expression levels and by associated metabolic noise distributions. To capture these biological variations unperturbed, highly sensitive in situ analytical techniques are needed that can sample tissue embedded single cells with minimum sample preparation. Optical fiber-based laser ablation electrospray ionization mass spectrometry (f-LAESI-MS) is a promising tool for metabolic profiling of single cells under ambient conditions. Integration of this MS-based platform with fluorescence and brightfield microscopy provides the ability to target single cells of specific type and allows for the selection of rare cells, e.g., excretory idioblasts. Analysis of individual Egeria densa leaf blade cells (n = 103) by f-LAESI-MS revealed significant differences between the prespecified subpopulations of epidermal cells (n = 97) and excretory idioblasts (n = 6) that otherwise would have been masked by the population average. Primary metabolites, e.g., malate, aspartate, and ascorbate, as well as several glucosides were detected in higher abundance in the epidermal cells. The idioblasts contained lipids, e.g., PG(16:0/18:2), and triterpene saponins, e.g., medicoside I and azukisaponin I, and their isomers. Metabolic noise for the epidermal cells were compared to results for soybean (Glycine max) root nodule cells (n = 60) infected by rhizobia (Bradyrhizobium japonicum). Whereas some primary metabolites showed lower noise in the latter, both cell types exhibited higher noise for secondary metabolites. Post hoc grouping of epidermal and root nodule cells, based on the abundance distributions for certain metabolites (e.g., malate), enabled the discovery of cellular subpopulations characterized by different mean abundance values, and the magnitudes of the corresponding metabolic noise. Comparison of prespecified populations from epidermal cells of the closely related E. densa (n = 20) and Elodea canadensis (n = 20) revealed significant differences, e.g., higher sugar content in the former and higher levels of ascorbate in the latter, and the presence of species-specific metabolites. These results demonstrate that the f-LAESI-MS single cell analysis platform has the potential to explore cellular heterogeneity and metabolic noise for hundreds of tissue-embedded cells.

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“…In the present review, we focused primarily our attention on the use of ME for drug discovery and development and in preclinical toxicology studies, its application for the analysis of bulk and single cells, for the detection of biomarkers in biological fluids, as well as for monitoring neurotransmitters in vitro and in vivo. For further discussion of applications and advances in ME (and CE) see the recent review written by Sibbitts et al [136] and Ragab and Kimary [137].…”

Section: Me For Separation and Quantification Of Neurotransmitters Inmentioning

confidence: 99%

Microfluidics as a Novel Tool for Biological and Toxicological Assays in Drug Discovery Processes: Focus on Microchip Electrophoresis

et al. 2020

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The last decades of biological, toxicological, and pharmacological research have deeply changed the way researchers select the most appropriate ‘pre-clinical model’. The absence of relevant animal models for many human diseases, as well as the inaccurate prognosis coming from ‘conventional’ pre-clinical models, are among the major reasons of the failures observed in clinical trials. This evidence has pushed several research groups to move more often from a classic cellular or animal modeling approach to an alternative and broader vision that includes the involvement of microfluidic-based technologies. The use of microfluidic devices offers several benefits including fast analysis times, high sensitivity and reproducibility, the ability to quantitate multiple chemical species, and the simulation of cellular response mimicking the closest human in vivo milieu. Therefore, they represent a useful way to study drug–organ interactions and related safety and toxicity, and to model organ development and various pathologies ‘in a dish’. The present review will address the applicability of microfluidic-based technologies in different systems (2D and 3D). We will focus our attention on applications of microchip electrophoresis (ME) to biological and toxicological studies as well as in drug discovery and development processes. These include high-throughput single-cell gene expression profiling, simultaneous determination of antioxidants and reactive oxygen and nitrogen species, DNA analysis, and sensitive determination of neurotransmitters in biological fluids. We will discuss new data obtained by ME coupled to laser-induced fluorescence (ME-LIF) and electrochemical detection (ME-EC) regarding the production and degradation of nitric oxide, a fundamental signaling molecule regulating virtually every critical cellular function. Finally, the integration of microfluidics with recent innovative technologies—such as organoids, organ-on-chip, and 3D printing—for the design of new in vitro experimental devices will be presented with a specific attention to drug development applications. This ‘composite’ review highlights the potential impact of 2D and 3D microfluidic systems as a fast, inexpensive, and highly sensitive tool for high-throughput drug screening and preclinical toxicological studies.

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Cellular Analysis Using Microfluidics

Cited by 51 publications

References 130 publications

Recent advances in direct current electrokinetic manipulation of particles for microfluidic applications

Recent advances in direct current electrokinetic manipulation of particles for microfluidic applications

Metabolic Noise and Distinct Subpopulations Observed by Single Cell LAESI Mass Spectrometry of Plant Cells in situ

Microfluidics as a Novel Tool for Biological and Toxicological Assays in Drug Discovery Processes: Focus on Microchip Electrophoresis

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