Pluronic Additives: A Solution to Sticky Problems in Digital Microfluidics

Luk, Vivienne N.; Mo, Gary; Wheeler, Aaron R.

doi:10.1021/la7039509

Cited by 198 publications

(199 citation statements)

References 43 publications

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“…In all DMF experiments, reagent solutions were supplemented with 0.02% Pluronics F68 except for sol-state Geltrex and collagen that were supplemented with 0.02% Pluronics F127 (ref. 56). Imaging during droplet manipulation was performed with a built-in webcam 55 .…”

Section: Methodsmentioning

confidence: 99%

Microgels on-demand

Eydelnant

Wheeler

2014

Nat Commun

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Three-dimensional (3D) hydrogel structures are finding use in fundamental studies of self-assembly, rheology, and 3D cell culture. Most techniques for 3D hydrogel formation are 'single pot', in which gels are not addressable after formation. For many applications, it would be useful to be able to form arrays of gels bearing mixtures of constituents and/or formed from composites of different gel materials. Here, in response to this challenge, we introduce a digital microfluidic method for 'on-demand' formation of arrays of microgels bearing arbitrary contents and shapes. On formation of the gels, each microgel is individually addressable for reagent delivery and analysis. We demonstrate the utility of the method for 3D cell culture and higher-order tissue formation by implementing the first sub-microlitre recapitulation of 3D kidney epithelialization. We anticipate this platform will enable new research that can exploit the flexible nature of this technique for forming and addressing arrays of hydrogels with unique geometries and contents.

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Section: Methodsmentioning

confidence: 99%

Microgels on-demand

Eydelnant

Wheeler

2014

Nat Commun

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“…In the work reported here, protein sticking was controlled by the use of low concentrations of amphiphilic polymer additives (chapter 2). 128 In other contexts, we have observed that these additives have little or no effect on biochemical processes ranging from cell vitality 71 to enzyme kinetics, 72 and can prevent adsorption even in highly concentrated, heterogeneous solutions such as serum (chapter 2). 128 The combination of these strategies, that is, incubation in humidified chambers and the use of adhesion-reducing additives, facilitated the development of these digital microfluidic techniques.…”

Section: Multistep Proteomic Processing On Dmf Devicesmentioning

confidence: 99%

“…50,71,72,128 In each assay, fluorescence from processed samples was measured in intervals of 10 min for 1 h. Between intervals, devices containing the reacting droplets were stored in a humidified chamber to prevent evaporation. Three replicate trials were conducted for each condition, and data were normalized and fitted with exponential functions.…”

Section: Reaction Rate Analysismentioning

confidence: 99%

A Digital Microfluidic Approach to Proteomic Sample Processing

2009

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Proteome profiling is the identification and quantitation of all proteins in biological samples. An important application of proteome profiling that has received much attention is clinical proteomics, a field that promises the discovery of biomarkers that will be useful for early diagnosis and prognosis of diseases. While clinical proteomic methods vary widely, a common characteristic is the need for (i) extraction of proteins from complex biological fluids and (ii) extensive biochemical processing (reduction, alkylation and enzymatic digestion) prior to analysis. However, the lack of standardized sample handling and processing in proteomics is a major limitation for the field. The conventional macroscale manual sample handling requires multiple containers and transfers, which often leads to sample loss and contamination. For clinical proteomics to be adopted as a gold standard for clinical measures, the issue of irreproducibility needs to be addressed. A potential solution to this problem is to form integrated systems for sample handling and processing, and in this dissertation, I describe my work towards realizing this goal using digital microfluidics (DMF). DMF is a technique characterized by the manipulation of discrete droplets (100 nL -10 L) on an array of electrodes by the application of electrical fields. It is well-suited for carrying out rapid, sequential, miniaturized automated biochemical assays. This thesis demonstrates how DMF can be a powerful tool capable of automating several protein handling and processing steps used in proteomics.iii Methods for implementing proteomic multi-step solution-phase processes (reduction, alkylation, and digestion) on DMF were developed. This was accompanied by the development of heterogenous enzymatic microreactors for efficient protein digestion. Strategies for reducing non-specific adsorption were developed. Finally, in proof-of-concept work, a combination of protein extraction and protein processing was demonstrated on DMF devices.iv ACKNOWLEDGEMENTS

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“…Encapsulation by immiscible oil is furthermore desired as it aids in controlling the rapid evaporation of nanoliter droplets. In a recent study, Luk et al 35 reported on the use of biocompatible polyethylene glycol ͑PEG͒ based additives in protein solutions to overcome biofouling on Teflon ® coated surfaces in electrowetting based microfluidic devices. 35 They showed that actuation of concentrated protein solutions ͑up to 50 mg/ml; bovine serum albumin ͑BSA͒, fibrinogen͒ is greatly facilitated ͑in air͒ by adding a small amount of PEG in protein solutions.…”

Section: Applicationsmentioning

confidence: 99%

Liquid dielectrophoresis and surface microfluidics

Kaler

Prakash²,

Chugh³

2010

Biomicrofluidics

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Liquid dielectrophoresis ͑L-DEP͒, when deployed at microscopic scales on top of hydrophobic surfaces, offers novel ways of rapid and automated manipulation of very small amounts of polar aqueous samples for microfluidic applications and development of laboratory-on-a-chip devices. In this article we highlight some of the more recent developments and applications of L-DEP in handling and processing of various types of aqueous samples and reagents of biological relevance including emulsions using such microchip based surface microfluidic ͑SMF͒ devices. We highlighted the utility of these devices for on-chip bioassays including nucleic acid analysis. Furthermore, the parallel sample processing capabilities of these SMF devices together with suitable on-or off-chip detection capabilities suggest numerous applications and utility in conducting automated multiplexed assays, a capability much sought after in the high throughput diagnostic and screening assays.

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Pluronic Additives: A Solution to Sticky Problems in Digital Microfluidics

Cited by 198 publications

References 43 publications

Microgels on-demand

Microgels on-demand

A Digital Microfluidic Approach to Proteomic Sample Processing

Liquid dielectrophoresis and surface microfluidics

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