Micro patterning of active proteins with perforated PDMS sheets (PDMS sieve)

Atsuta, Kyoko; Noji, Hiroyuki; Takeuchi, Shoji

doi:10.1039/b400774c

Cited by 39 publications

(30 citation statements)

References 20 publications

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“…[9–11] In addition to direct microprinting technologies, [12, 13] a number of photolithographic techniques have been designed to obtain micro-patterns of biomolecules on a variety of substrates. [14–16] Patterning of multiple different proteins has also been demonstrated by soft lithography, [13, 17, 18] stencils [19–21] and other methods. [22–24] …”

Section: Introductionmentioning

confidence: 99%

Affinity‐Based Protein Surface Pattern Formation by Ligand Self‐Selection from Mixed Protein Solutions

Dubey

Emoto

Takahashi

et al. 2009

Adv Funct Materials

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Photolithographically prepared surface patterns of two affinity ligands (biotin and chloroalkane) specific for two proteins (streptavidin and HaloTag®, respectively) are used to spontaneously form high-fidelity surface patterns of the two proteins from their mixed solution. High affinity protein-surface self-selection onto patterned ligands on surfaces exhibiting low non-specific adsorption rapidly yields the patterned protein surfaces. Fluorescence images after protein immobilization show high specificity of the target proteins to their respective surface patterned ligands. Time-of-flight secondary ion mass spectrometry (ToF-SIMS) imaging further supports the chemical specificity of streptavidin and HaloTag® for their surface patterned ligands from mixed protein solutions. However, ToF-SIMS did detect some non-specific adsorption of bovine serum albumin, a masking protein present in excess in the adsorbing solutions, on the patterned surfaces. Protein amino acid composition, surface coverage, density and orientation are important parameters that determine the relative ToF-SIMS fragmentation pattern yields. ToF-SIMS amino acid-derived ion fragment yields summed to produce surface images can reliably determine which patterned surface regions contain bound proteins, but do not readily discriminate between different co-planar protein regions. Principal component analysis (PCA) of these ToF-SIMS data, however, improves discrimination of ions specific to each protein, facilitating surface pattern discrimination and contrast.

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Section: Introductionmentioning

confidence: 99%

Affinity‐Based Protein Surface Pattern Formation by Ligand Self‐Selection from Mixed Protein Solutions

Dubey

Emoto

Takahashi

et al. 2009

Adv Funct Materials

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“…The technology further allows for the generation of dynamic co‐patterns of multiple proteins. Selective patterning of proteins such as antibodies and enzymes, for example, has recently attracted much interest for the study of specific protein–protein interactions and the development of diagnostic kits, protein sensors, and protein chips 32–35. Parylene‐C technology might find application in this rapidly developing field of selective protein patterning.…”

Section: Resultsmentioning

confidence: 99%

Microfabricated multilayer parylene‐C stencils for the generation of patterned dynamic co‐cultures

Jinno

Moeller

Chen

et al. 2008

J Biomedical Materials Res

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Co-culturing different cell types can be useful to engineer a more in vivo-like microenvironment for cells in culture. Recent approaches to generating cellular co-cultures have used microfabrication technologies to regulate the degree of cell-cell contact between different cell types. However, these approaches are often limited to the co-culture of only two cell types in static cultures. The dynamic aspect of cell-cell interaction, however, is a key regulator of many biological processes such as early development, stem cell differentiation, and tissue regeneration. In this study, we describe a micropatterning technique based on microfabricated multilayer parylene-C stencils and demonstrate the potential of parylene-C technology for co-patterning of proteins and cells with the ability to generate a series of at least five temporally controlled patterned co-cultures. We generated dynamic co-cultures of murine embryonic stem cells in culture with various secondary cell types that could be sequentially introduced and removed from the co-cultures. Our studies suggested that dynamic co-cultures generated by using parylene-C stencils may be applicable in studies investigating cellular interactions in controlled microenvironments such as studies of ES cell differentiation, wound healing and development.

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“…Several microstructuring techniques have been developed to manufacture thin PDMS membranes with µm-sized pores 52–57 . A technique that is easy to use and yields pores down to a few µm in diameter is to spin coat PDMS pre-polymer on molds that bear thin posts in order to generate the pores 54, 56, 57 .…”

Section: Microfluidic Biochip Architectures For Culturing Epitheliamentioning

confidence: 99%

“…Although this technique is more difficult to use, well-defined pores down to 1 µm in diameter can be produced. Furthermore, methods to manufacture pores in PDMS membranes with sub-µm diameter have been described 52 . To this end, PDMS pre-polymer was spin coated on a mold with pyramid-shaped posts.…”

Section: Microfluidic Biochip Architectures For Culturing Epitheliamentioning

confidence: 99%

Microfluidic approaches for epithelial cell layer culture and characterisation

Thuenauer

Rodríguez-Boulan

Römer

2014

Analyst

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In higher eukaryotes, epithelial cell layers line most body cavities and form selective barriers that regulate the exchange of solutes between compartments. In order to fulfil these functions, the cells assume a polarised architecture and maintain two distinct plasma membrane domains, the apical domain facing the lumen and the basolateral domain facing other cells and the extracellular matrix. Microfluidic biochips offer the unique opportunity to establish novel in vitro models of epithelia in which the in vivo microenvironment of epithelial cells is precisely reconstituted. In addition, analytical tools to monitor biologically relevant parameters can be directly integrated on-chip. In this review we summarise recently developed biochip designs for culturing epithelial cell layers. Since endothelial cell layers, which line blood vessels, have similar barrier functions and polar organisation as epithelial cell layers, we also discuss biochips for culturing endothelial cell layers. Furthermore, we review approaches to integrate tools to analyse and manipulate epithelia and endothelia in microfluidic biochips, including methods to perform electrical impedance spectroscopy, methods to detect substances undergoing trans-epithelial transport via fluorescence, spectrophotometry, and mass spectrometry, techniques to mechanically stimulate cells via stretching and fluid flow-induced shear stress, and methods to carry out high-resolution imaging of vesicular trafficking with light microscopy. Taken together, this versatile microfluidic toolbox enables novel experimental approaches to characterise epithelial monolayers.

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Micro patterning of active proteins with perforated PDMS sheets (PDMS sieve)

Cited by 39 publications

References 20 publications

Affinity‐Based Protein Surface Pattern Formation by Ligand Self‐Selection from Mixed Protein Solutions

Affinity‐Based Protein Surface Pattern Formation by Ligand Self‐Selection from Mixed Protein Solutions

Microfabricated multilayer parylene‐C stencils for the generation of patterned dynamic co‐cultures

Microfluidic approaches for epithelial cell layer culture and characterisation

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