Multiprotein Microcontact Printing with Micrometer Resolution

Eichinger, Colin D.; Hsiao, Tony W.; Hlady, Vladimir

doi:10.1021/la2039202

Cited by 31 publications

(32 citation statements)

References 28 publications

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“…The pattern used in this study consisted of 15 μm stripes separated by 25 μm–wide gaps. This pattern was selected as 15 μm stripes previously aligned astrocytes on glass [27], and the added asymmetry was useful to distinguish printed and non-printed regions while maintaining alignment [36]. In the second step, a process was developed for collagen to undergo simultaneous gelling and cross-linking while the protein patterns were transferred (Figure 1).…”

Section: Methodsmentioning

confidence: 99%

“…Astrocyte alignment was assessed by measuring the orientation of their nuclei [27,36]. To do this, the angle of the major axis of the oval-like astrocyte nuclei were quantified using the measure angle function of Image Pro Plus.…”

Section: Methodsmentioning

confidence: 99%

“…However, to the best of our knowledge, the present report is the first application of protein pattern transfer and covalent attachment to the surface of collagen type I hydrogels and requires no additional, specialized equipment beyond standard materials for creating protein patterns. The techniques described here will be conducive to the transfer of multiple proteins deposited in any desired pattern on glass surface by traditional microcontact printing [34], stamp-off [35], sequential stamping, microscope-aided registration [36], dip-pen nanolithography [37], or inkjet deposition [38], directly to collagen hydrogel surface.…”

Section: Introductionmentioning

confidence: 99%

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Astrocytes alignment and reactivity on collagen hydrogels patterned with ECM proteins

2015

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To modulate the surface properties of collagen and subsequent cell-surface interactions, a method was developed to transfer protein patterns from glass coverslips to collagen type I hydrogel surfaces. Two proteins and one proteoglycan found in central nervous system extracellular matrix as well as fibrinogen were patterned in stripes onto collagen hydrogel and astrocytes were cultured on these surfaces. The addition of the stripe protein patterns to hydrogels created astrocyte layers in which cells were aligned with underlying patterns and had reduced chondroitin sulfate expression compared to the cells grown on collagen alone. Protein patterns were covalently cross-linked to the collagen and stable over four days in culture with no visible cellular modifications. The present method can be adapted to transfer other types of protein patterns from glass coverslips to collagen hydrogels.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Astrocytes alignment and reactivity on collagen hydrogels patterned with ECM proteins

2015

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“…For instance, it has been found that the size and geometry of the accessible area can alter and direct the axis of cell polarization and division 2,3,5 , the positions and orientation of pseudopodia 4 , or the locations of junctions between adjacent cells 14 . Cell adhesion and migration depends in large part on the protein composition of a cell's surroundings [15][16][17] , but the relative density of adhesion sites [18][19][20] , as well as density gradients of surface-bound proteins, can influence and bias its spreading and motion 21,22 .…”

Section: Introductionmentioning

confidence: 99%

Versatile method to generate multiple types of micropatterns

et al. 2016

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Micropatterning techniques have become an important tool for the study of cell behavior in controlled microenvironments. As a consequence, several approaches for the creation of micropatterns have been developed in recent years. However, the diversity of substrates, coatings and complex patterns used in cell science is so great that no single existing technique is capable of fabricating designs suitable for all experimental conditions. Hence, there is a need for patterning protocols that are flexible with regard to the materials used and compatible with different patterning strategies to create more elaborate setups. In this work, we present a versatile approach to micropatterning. The protocol is based on plasma treatment, protein coating, and a PLL-PEG backfill step, and produces homogeneous patterns on a variety of substrates. Protein density within the patterns can be controlled, and density gradients of surface-bound protein can be formed. Moreover, by combining the method with microcontact printing, it is possible to generate patterns composed of three different components within one iteration of the protocol. The technique is simple to implement and should enable cell science labs to create a broad range of complex and highly specialized microenvironments.

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“…Another approach to patterning is direct printing (ink jet), which can produce spot patterns with a resolution of approximately 200 lm. 15,16 Other variations for studying interactions include contact printing, [17][18][19] 24 These and other techniques have been reviewed in the literature. 25 Stencil approaches have used fluidic patterning with dry liftoff or surface chemical patterning to form multi-component, bio-molecular patterns at micron-scale resolution.…”

Section: Introductionmentioning

confidence: 99%

Microstencils to generate defined, multi-species patterns of bacteria

et al. 2015

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Microbial communities are complex heterogeneous systems that are influenced by physical and chemical interactions with their environment, host, and community members. Techniques that facilitate the quantitative evaluation of how microscale organization influences the morphogenesis of multispecies communities could provide valuable insights into the dynamic behavior and organization of natural communities, the design of synthetic environments for multispecies culture, and the engineering of artificial consortia. In this work, we demonstrate a method for patterning microbes into simple arrangements that allow the quantitative measurement of growth dynamics as a function of their proximity to one another. The method combines parylene-based liftoff techniques with microfluidic delivery to simultaneously pattern multiple bacterial species with high viability using low-cost, customizable methods. Quantitative measurements of bacterial growth for two competing isolates demonstrate that spatial coordination can play a critical role in multispecies growth and structure. V C 2015 AIP Publishing LLC. [http://dx

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Multiprotein Microcontact Printing with Micrometer Resolution

Cited by 31 publications

References 28 publications

Astrocytes alignment and reactivity on collagen hydrogels patterned with ECM proteins

Astrocytes alignment and reactivity on collagen hydrogels patterned with ECM proteins

Versatile method to generate multiple types of micropatterns

Microstencils to generate defined, multi-species patterns of bacteria

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