Living-Cell Microarrays

Yarmush, Martin L.; King, Kevin R.

doi:10.1146/annurev.bioeng.10.061807.160502

Cited by 122 publications

(128 citation statements)

References 117 publications

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“…Microarray technique allows scanning a number of images (Fernandes et al 2009;Yarmush and King 2009) and microfluidics (Yin and Marshall 2012) can be used to provide reagent addition and realize reactions in miniature volumes operating with single cells or small number of cells. Fluorescence spectroscopy and flow cytometry are achievable on microscale for providing in-chip analysis of apoptosis (Martinez et al 2010a).…”

Section: Fluorescence Techniques In the Detection Of Apoptosismentioning

confidence: 99%

Beyond annexin V: fluorescence response of cellular membranes to apoptosis

2012

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Dramatic changes in the structure of cell membranes on apoptosis allow easy, sensitive and non-destructive analysis of this process with the application of fluorescence methods. The strong plasma membrane asymmetry is present in living cells, and its loss on apoptosis is commonly detected with the probes interacting strongly and specifically with phosphatidylserine (PS). This phospholipid becomes exposed to the cell surface, and the application of annexin V labeled with fluorescent dye is presently the most popular tool for its detection. Several methods have been suggested recently that offer important advantages over annexin V assay with the ability to study apoptosis by spectroscopy of cell suspensions, flow cytometry and confocal or twophoton microscopy. The PS exposure marks the integrated changes in the outer leaflet of cell membrane that involve electrostatic potential and hydration, and the attempts are being made to provide direct probing of these changes. This review describes the basic mechanisms underlying the loss of membrane asymmetry during apoptosis and discusses, in comparison with the annexin V-binding assay, the novel fluorescence techniques of detecting apoptosis on cellular membrane level. In more detail we describe the detection method based on smart fluorescent dye F2N12S incorporated into outer leaflet of cell membrane and reporting on apoptotic cell transformation by easily detectable change of the spectral distribution of fluorescent emission. It can be adapted to any assay format.

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Section: Fluorescence Techniques In the Detection Of Apoptosismentioning

confidence: 99%

Beyond annexin V: fluorescence response of cellular membranes to apoptosis

2012

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“…cell array | cell communication | protrusion profiling | neuron patterning C urrent high-throughput screening of cell function and heterogeneity and in vitro cell-cell communication studies requires routine generation of large-scale single-cell arrays with high precision and efficiency, single-cell resolution, multiple cell types, and maintenance of cell viability and function (1,2). Several approaches have been designed for this purpose, e.g., inkjet cell printing (3)(4)(5)(6), surface engineering (7)(8)(9)(10)(11)(12)(13)(14)(15), and physical constraints (16)(17)(18)(19)(20)(21)(22)(23).…”

mentioning

confidence: 99%

Block-Cell-Printing for live single-cell printing

Zhang¹,

Chou

Xia

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

127

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A unique live-cell printing technique, termed "Block-Cell-Printing" (BloC-Printing), allows for convenient, precise, multiplexed, and high-throughput printing of functional single-cell arrays. Adapted from woodblock printing techniques, the approach employs microfluidic arrays of hook-shaped traps to hold cells at designated positions and directly transfer the anchored cells onto various substrates. BloC-Printing has a minimum turnaround time of 0.5 h, a maximum resolution of 5 μm, close to 100% cell viability, the ability to handle multiple cell types, and efficiently construct protrusion-connected single-cell arrays. The approach enables the large-scale formation of heterotypic cell pairs with controlled morphology and allows for material transport through gap junction intercellular communication. When six types of breast cancer cells are allowed to extend membrane protrusions in the BloC-Printing device for 3 h, multiple biophysical characteristics of cells-including the protrusion percentage, extension rate, and cell length-are easily quantified and found to correlate well with their migration levels. In light of this discovery, BloC-Printing may serve as a rapid and high-throughput cell protrusion characterization tool to measure the invasion and migration capability of cancer cells. Furthermore, primary neurons are also compatible with BloC-Printing.cell array | cell communication | protrusion profiling | neuron patterning C urrent high-throughput screening of cell function and heterogeneity and in vitro cell-cell communication studies requires routine generation of large-scale single-cell arrays with high precision and efficiency, single-cell resolution, multiple cell types, and maintenance of cell viability and function (1, 2). Several approaches have been designed for this purpose, e.g., inkjet cell printing (3-6), surface engineering (7-15), and physical constraints (16-23). However, finding a method that completely satisfies the above requirements remains a challenge. Potentially useful and convenient tools may be available by adapting traditional printing tools to cell printing. In particular, woodblock printing is an efficient and convenient technology that revolutionized the printing world more than 1,800 y ago and was extended to microcontact molecular printing ∼20 y ago (24). However, application of the block-printing concept to cells has never been achieved. The main challenges are (i) inking cells to their molds with precision and maintaining viability, (ii) evenly and gently applying and transferring the cells to a substrate and successfully lifting off the mold without detaching the cells, and (iii) maintaining cell functions after printing.We report here the development and testing of a technology called "Block-Cell-Printing" (BloC-Printing), which involves directly inking cells to a predesigned mold and then transferring the cells to a substrate. We overcome the challenges described above by flow patterning, instead of pressing, the ink objects, as in woodblock or microcontact printing. By ...

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“…These include real-time studies on a single cell level, such as timeof-flight (TOF) optophoresis and scanning thermal lens microscopy (TLM) [46][47][48][49][50] . The living cell microarrays and microfluidic cell arrays are yet other examples of emerging LOC technologies that provide important technological advances in the spatiotemporal control of biomolecules and cells [6,10,[51][52][53][54][55][56] . Pioneering microfluidic cell arrays allow collection of real-time and multiparameter data obtained from functional cell-based assays [6,10,56] .…”

Section: Emerging Prospect Of Microfluidics For Cancer Researchmentioning

confidence: 99%

Microfluidics: Emerging prospects for anti-cancer drug screening

Wlodkowic¹

2010

WJCO

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Cancer constitutes a heterogenic cellular system with a high level of spatio-temporal complexity. Recent discoveries by systems biologists have provided emerging evidence that cellular responses to anti-cancer modalities are stochastic in nature. To uncover the intricacies of cell-to-cell variability and its relevance to cancer therapy, new analytical screening technologies are needed. The last decade has brought forth spectacular innovations in the field of cytometry and single cell cytomics, opening new avenues for systems oncology and high-throughput real-time drug screening routines. The up-and-coming microfluidic Lab-on-a-Chip (LOC) technology and micrototal analysis systems (µTAS) are arguably the most promising platforms to address the inherent complexity of cellular systems with massive experimental parallelization and 4D analysis on a single cell level. The vast miniaturization of LOC systems and multiplexing enables innovative strategies to reduce drug screening expenditures while increasing throughput and content of information from a given sample. Small cell numbers and operational reagent volumes are sufficient for microfluidic analyzers and, as such, they enable next generation high-throughput and high-content screening of anticancer drugs on patient-derived specimens. Herein we highlight the selected advancements in this emerging field of bioengineering, and provide a snapshot of developments with relevance to anti-cancer drug screening routines.

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Living-Cell Microarrays

Cited by 122 publications

References 117 publications

Beyond annexin V: fluorescence response of cellular membranes to apoptosis

Beyond annexin V: fluorescence response of cellular membranes to apoptosis

Block-Cell-Printing for live single-cell printing

Microfluidics: Emerging prospects for anti-cancer drug screening

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