Patterned Protein Layers on Solid Substrates by Thin Stamp Microcontact Printing

James, Conrad D.; Davis, R.; Kam, Lance C.; Craighead, Harold G.; Isaacson, M.; Turner, James N.; Shain, William

doi:10.1021/la9710482

Cited by 275 publications

(205 citation statements)

References 14 publications

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“…PDMS layers between 220 µm and 1 mm thick were able form a good seal with the polystyrene substrate but were not so thick that the microchannels collapsed. 13 …”

Section: Experimental Section Design and Fabrication Of Capillary Masksmentioning

confidence: 99%

Catch Strip Assay for the Relative Assessment of Two-Dimensional Protein Association Kinetics

et al. 2008

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Accurate interpretation of recruitment rate measurements of microscale particles, such as cells and microbeads, to biofunctional surfaces is difficult because factors such as uneven ligand distributions, particle collisions, variable particle fluxes, and molecular-scale surface separation distances obfuscate the ability to link the observed particle behavior with the governing nanoscale biophysics. We report the development of a hydrodynamically conditioned micropattern catch strip assay to measure microparticle recruitment kinetics. The assay exploited patterning within microfluidic channels and the mechanostability of selectin bonds to create reaction geometries that confined a microbead flux to within 200 nm of the surface under flow conditions. Systematic control of capillary action enabled the creation of homogeneous or gradient ligand distributions. The method enabled the measurement of particle recruitment rates (k eff , s −1 ) that were primarily determined by the interaction of the biomolecular pair being investigated. The method is therefore well suited for relative measurements of delivery vehicle and cellular recruitment potential as governed by surface-bound molecules.The scientific and clinical importance of understanding the recruitment of leukocytes, stem cells, metastatic cancer cells, medical imaging contrast agents, and drug delivery vehicles to tissues has led to the development of a number of assays to analyze each stage in the process. A frequently exploited technology is the in vitro flow chamber assay. Studies of adhesion employing flow chambers require culturing cells 1 or immobilizing ligands on a target surface, 2,3 flowing a suspension of cells over the surface and observing interactions. Despite their inherent complexity, flow chamber assays have led to quantitative insights into some of the biophysical factors important for cell adhesion, especially in the context of cellular dissociation rates. 4 Although the flow chamber assay has been used successfully to deduce the apparent bond lifetimes of mechanically stressed cell adhesion molecules, several factors have made the determination of a molecularly mediated capture parameter intrinsic to the microparticle more difficult. For one, the flux of microparticles sufficiently close to the surface to engage in adhesive interactions changes with the wall shear rate and position in the flow chamber. 5,6 In addition, factors such as the hydrodynamic effects of bound microparticles on the flow © 2008 American Chemical Society * To whom correspondence should be addressed. mbl2a@virginia.edu. SUPPORTING INFORMATION AVAILABLEA movie demonstration of the assay, functional analysis of ligand distribution, analysis of applicability of the Poisson distribution, AFM results, and additional details on the patterning, tracking, and statistical methodology. This material is available free of charge via the Internet at http://pubs.acs.org. We developed an assay that geometrically controlled the presentation of adhesive ligand to establish a defi...

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“…PDMS layers between 220 µm and 1 mm thick were able form a good seal with the polystyrene substrate but were not so thick that the microchannels collapsed. 13 …”

Section: Experimental Section Design and Fabrication Of Capillary Masksmentioning

confidence: 99%

Catch Strip Assay for the Relative Assessment of Two-Dimensional Protein Association Kinetics

et al. 2008

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“…Resists made of SAMs can also be used as masks for isotropic wet etchants on a limited set of substrates (mainly, Originally demonstrated for alkanethiol inks transferred on gold for producing patterns of SAMs, 11 the principle works also for the transfer of dried proteins from PDMS to various surfaces, as demonstrated by three groups simultaneously. [23][24][25] Although microstamping of proteins represents a protein immobilization protocol (based on contact transfer of a dry layer) that departs substantially from the traditional one (based on deposition from solution), it is experimentally simple and allows for greater flexibility and resolution than microfluidic patterning. Alignment of two non-overlapping protein patterns is facilitated by the use of a single multilevel stamp.…”

Section: Iib Micromachining (Etching and Deposition)mentioning

confidence: 99%

“…To study cell contact guidance in simultaneous presence of a topographic and an adhesive cue, Britland et al 90 presented BHK fibroblasts with grooves of various depths (0.1, 0.5, 1.0, 3.0, and 6.0 μm deep) together with pitch-matched aminosilane tracks of various widths (5,12,25,50, and 100 μm wide) at parallel and orthogonally opposed directions. They found that cell alignment was profoundly enhanced on all surfaces that presented both cues in parallel; cells were able to switch alignment from ridges to grooves, and vice versa, depending on the location of superimposed adhesive tracks; cells aligned preferentially to adhesive tracks superimposed orthogonally over grooves of matched pitch, traversing numerous grooves and ridges.…”

Section: Effect Of Microtopography On Fibroblastmentioning

confidence: 99%

Biology on a Chip: Microfabrication for Studying the Behavior of Cultured Cells

Tourovskaia

Folch

2003

Crit Rev Biomed Eng

173

140

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The ability to culture cells in vitro has revolutionized hypothesis testing in basic cell and molecular biology research and has become a standard methodology in drug screening and toxicology assays. However, the traditional cell culture methodology-consisting essentially of the immersion of a large population of cells in a homogeneous fluid medium-has become increasingly limiting, both from a fundamental point of view (cells in vivo are surrounded by complex spatiotemporal microenvironments) and from a practical perspective (scaling up the number of fluid handling steps and cell manipulations for high-throughput studies in vitro is prohibitively expensive). Micro fabrication technologies have enabled researchers to design, with micrometer control, the biochemical composition and topology of the substrate, the medium composition, as well as the type of neighboring cells surrounding the microenvironment of the cell. In addition, microtechnology is conceptually well suited for the development of fast, low-cost in vitro systems that allow for high-throughput culturing and analysis of cells under large numbers of conditions. Here we review a variety of applications of microfabrication in cell culture studies, with an emphasis on the biology of various cell types.

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“…In most cases, nearly 100% of the protein film transfers from the stamp onto the surface being patterned, presumably due to favorable interactions between the exposed protein film surface and the substrate to which the film is transferred. Some printed protein fields have also shown largely unaltered epitope presentation in antibody immunostaining assays [107,108]. It has been demonstrated that stamped protein fields retain sufficient activity to promote the levels of cell adhesion and phenotypic expression found for the same cell-protein systems where the protein was deposited by adsorption.…”

Section: Protein μCp Stamping and μFn Depositionmentioning

confidence: 99%

The effects of proteoglycan surface patterning on neuronal pathfinding

Hlady

Hodgkinson

2007

Materialwissenschaft Werkst

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Protein micropatterning techniques are increasingly applied in cell choice assays to investigate fundamental biological phenomena that contribute to the host response to implanted biomaterials, and to explore the effects of protein stability and biological activity on cell behavior for in vitro cell studies. In the area of neuronal regeneration the protein micropatterning and cell choice assays are used to improve our understanding of the mechanisms directing nervous system during development and regenerative failure in the central nervous system (CNS) wound healing environment. In these cell assays, protein micropatterns need to be characterized for protein stability, bioactivity, and spatial distribution and then correlated with observed mammalian cell behavior using appropriate model system for CNS development and repair. This review provides the background on protein micropatterning for cell choice assays and describes some novel patterns that were developed to interrogate neuronal adaptation to inhibitory signals encountered in CNS injuries. Rationale for studying the role of proteoglycans in CNS injuriesNeurons from the central nervous system (CNS) possess a limited capacity to regenerate beyond scar tissue formation barriers in injuries even in the presence of minimal trauma to tissue organization [1,2]. A major culprit in CNS neuronal regenerative failure are the inihibitory proteoglycans (PGs), which are upregulated at the site of CNS injuries. PGs are composed of a core protein with varying numbers of attached glycosaminoglycan (GAG) side chains [3,4]. Proteoglycans are first translated intracellularly into proteins in the endoplasmic reticulum, and then GAG chains are later attached in the Golgi apparatus before secretion into the extracellular space [5][6][7][8]. The study of PGs has particularly been pronounced in the field of neurobiology and development of the nervous system. Numerous studies have concluded that PGs act as barriers that direct the migration of neural crest cells and the outgrowth direction of pathfinding neurons during development [9][10][11][12][13][14][15][16][17]. A number of these studies have additionally shown that cell guidance by PGs is based on an inhibitory signal located within the chondroitin sulfate GAG chains [9,18,19], with the result that the artificial addition of chondroitin sulfate (CS) sugars to the developing nervous system disrupts normal pathfinding [20] as well as does the removal of chondroitin sulfate chains through enzymatic treatment [9,19]. Davies et al. found that chondroitin sulfate proteoglycans (CSPG) are a major constituent of the glial scar and that dorsal root ganglion (DRG) outgrowth termination coincided with an increase in CSPG expression density [21]. Other sources have also shown that CSPGs are upregulated after CNS injuries [22][23][24][25].Interestingly, native adult CNS neurons actually posses the intrinsic ability to regenerate when the wound healing environment is prepared by eliminating inhibitory factors [23,26,27]. In additi...

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Patterned Protein Layers on Solid Substrates by Thin Stamp Microcontact Printing

Cited by 275 publications

References 14 publications

Catch Strip Assay for the Relative Assessment of Two-Dimensional Protein Association Kinetics

Catch Strip Assay for the Relative Assessment of Two-Dimensional Protein Association Kinetics

Biology on a Chip: Microfabrication for Studying the Behavior of Cultured Cells

The effects of proteoglycan surface patterning on neuronal pathfinding

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