Microfluidic single-cell analysis of intracellular compounds

Chao, Tzu-Chiao; Ros, Alexandra

doi:10.1098/rsif.2008.0233.focus

Cited by 92 publications

(56 citation statements)

References 75 publications

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“…If 10 encoded microbeads are used to interrogate a protein species of interest to provide 10-fold oversampling, 250 different protein species can be analysed within this 100 Â 100 mm area, the typical size of one spot used in conventional printed microarrays [16,45]. Therefore, our method will enable the integration of antibody array fabrication into microfluidic devices for sensitive multiplexed protein analysis [23,39,41,46] and potentially for single cell analysis.…”

Section: Multiplexed Protein Immunoassays and Sensitivitymentioning

confidence: 99%

Multiplexed protein analysis using encoded antibody-conjugated microbeads

Theilacker

Roller

Barbee

et al. 2011

J. R. Soc. Interface.

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We describe a method for multiplexed analysis of proteins using fluorescently encoded microbeads. The sensitivity of our method is comparable to the sensitivity obtained by enzyme-linked immunosorbent assay while only 5 ml sample volumes are needed. Streptavidin-coated, 1 mm beads are encoded with a combination of fluorophores at different intensity levels. As a proof of concept, we demonstrate that 27 microbead populations can be readily encoded by affinity conjugation using three intensity levels for each of three different biotinylated fluorescent dyes. Four populations of encoded microbeads are further conjugated with biotinylated capture antibodies and then combined and immobilized in a microfluidic flow cell for multiplexed protein analysis. Using four uniquely encoded microbead populations, we show that a cancer biomarker and three cytokine proteins can be analysed quantitatively in the picogram per millilitre range by fluorescence microscopy in a single assay. Our method will allow for the fabrication of high density, bead-based antibody arrays for multiplexed protein analysis using integrated microfluidic devices and automated sample processing.

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Section: Multiplexed Protein Immunoassays and Sensitivitymentioning

confidence: 99%

Multiplexed protein analysis using encoded antibody-conjugated microbeads

Theilacker

Roller

Barbee

et al. 2011

J. R. Soc. Interface.

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“…A number of modern techniques have been developed for SCA including fluorescence methods, capillary electrophoresis, microfluidics, mass spectroscopy, etc. [4,5]. Mostly, these techniques are advantageous due to analysis of small volumes and due to increased sensitivity of analyte detection (e.g., by microfluidics).…”

Section: Introductionmentioning

confidence: 99%

Polyelectrolyte Multilayers: Towards Single Cell Studies

2014

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Single cell analysis (SCA) is nowadays recognized as one of the key tools for diagnostics and fundamental cell biology studies. The Layer-by-layer (LbL) polyelectrolyte assembly is a rather new but powerful technique to produce multilayers. It allows to model the extracellular matrix in terms of its chemical and physical properties. Utilization of the multilayers for SCA may open new avenues in SCA because of the triple role of the multilayer film: (i) high capacity for various biomolecules; (ii) natural mimics of signal molecule diffusion to a cell and (iii) cell patterning opportunities. Besides, light-triggered release from multilayer films offers a way to deliver biomolecules with high spatio-temporal resolution. Here we review recent works showing strong potential to use multilayers for SCA and address accordingly the following issues: biomolecule loading, cell patterning, and light-triggered release.

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“…In addition, it enables the experimental conditions parallelization, the measurement of cellular dynamics in real-time, and the delivery of reagents, nutrients, and other stimuli to cells precisely. [28][29][30][31] Particularly, it facilitates the recreation of the micro-scale cell confinement with critical cell-cell interfaces, spatiotemporal chemical gradients, and dynamic mechanical microenvironment of cells with a higher similarity to physiological conditions, which is not possible by conventional approach. These new capabilities that emerged from the convergence of microfluidics with microengineering and cell biology have led to the progresses to reconstitute the physiologically relevant tumor microenvironment on a chip.…”

Section: Introductionmentioning

confidence: 99%

Biomimetic tumor microenvironment on a microfluidic platform

Qin

2013

Biomicrofluidics

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Tumor microenvironment is a highly complex system consisting of non-cancerous cells, soluble factors, signaling molecules, extracellular matrix, and mechanical cues, which provides tumor cells with integrated biochemical and biophysical cues. It has been recognized as a significant regulator in cancer initiation, progression, metastasis, and drug resistance, which is becoming a crucial component of cancer biology. Modeling microenvironmental conditions of such complexity in vitro are particularly difficult and technically challenging. Significant advances in microfluidic technologies have offered an unprecedented opportunity to closely mimic the physiological microenvironment that is normally encountered by cancer cells in vivo. This review highlights the recent advances of microfluidic platform in recapitulating many aspects of tumor microenvironment from biochemical and biophysical regulations. The major events relevant in tumorigenesis, angiogenesis, and spread of cancer cells dependent on specific combinations of cell types and soluble factors present in microenvironmental niche are summarized. The questions and challenges that lie ahead if this field is expected to transform the future cancer research are addressed as well. V C 2013 American Institute of Physics.

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Microfluidic single-cell analysis of intracellular compounds

Cited by 92 publications

References 75 publications

Multiplexed protein analysis using encoded antibody-conjugated microbeads

Multiplexed protein analysis using encoded antibody-conjugated microbeads

Polyelectrolyte Multilayers: Towards Single Cell Studies

Biomimetic tumor microenvironment on a microfluidic platform

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