Independently Controlling Protein Dot Size and Spacing in Particle Lithography

Taylor, Zachary R.; Keay, Joel C.; Sanchez, Ernest S.; Johnson, Matthew B.; Schmidtke, David W.

doi:10.1021/la300806m

Cited by 21 publications

(17 citation statements)

References 46 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The shape of the pattern motif is a circle and is determined by a combination of bead-size and the duration of aluminum deposition 9,13 . Alternative techniques for controlling the dot size have also been suggested 14,17,18 .…”

Section: Discussionmentioning

confidence: 99%

Ligand Nano-cluster Arrays in a Supported Lipid Bilayer

Benard

Ozerov

et al. 2017

JoVE

View full text Add to dashboard Cite

Currently there is considerable interest in creating ordered arrays of adhesive protein islands in a sea of passivated surface for cell biological studies. In the past years, it has become increasingly clear that living cells respond, not only to the biochemical nature of the molecules presented to them but also to the way these molecules are presented. Creating protein micro-patterns is therefore now standard in many biology laboratories; nano-patterns are also more accessible. However, in the context of cell-cell interactions, there is a need to pattern not only proteins but also lipid bilayers. Such dual proteo-lipidic patterning has so far not been easily accessible. We offer a facile technique to create protein nano-dots supported on glass and propose a method to backfill the inter-dot space with a supported lipid bilayer (SLB). From photo-bleaching of tracer fluorescent lipids included in the SLB, we demonstrate that the bilayer exhibits considerable in-plane fluidity. Functionalizing the protein dots with fluorescent groups allows us to image them and to show that they are ordered in a regular hexagonal lattice. The typical dot size is about 800 nm and the spacing demonstrated here is 2 microns. These substrates are expected to serve as useful platforms for cell adhesion, migration and mechano-sensing studies.

show abstract

Section: Discussionmentioning

confidence: 99%

Ligand Nano-cluster Arrays in a Supported Lipid Bilayer

Benard

Ozerov

et al. 2017

JoVE

View full text Add to dashboard Cite

show abstract

“…In colloid lithography, polystyrene spheres in solution are deposited on a surface, and as the solvent evaporates, the spheres form a regular, packed monolayer (Taylor et al, 2012 ) (Figure 6 A). Spheres are then deformed by exposing them to a temperature gradient, which melts the spheres differentially based on the heat applied.…”

Section: Methods To Create Substrate-bound Protein Gradientsmentioning

confidence: 99%

“…(A) By applying a heat gradient under the surface, microspheres melt and interact with the surface to different extents thereby creating a gradient of dots of different size. Adapted with permission from Taylor et al ( 2012 ). Copyright 2012 American Chemical Society.…”

Section: Methods To Create Substrate-bound Protein Gradientsmentioning

confidence: 99%

Substrate-Bound Protein Gradients to Study Haptotaxis

Ricoult

Kennedy

Juncker

2015

Front. Bioeng. Biotechnol.

View full text Add to dashboard Cite

Cells navigate in response to inhomogeneous distributions of extracellular guidance cues. The cellular and molecular mechanisms underlying migration in response to gradients of chemical cues have been investigated for over a century. Following the introduction of micropipettes and more recently microfluidics for gradient generation, much attention and effort was devoted to study cellular chemotaxis, which is defined as guidance by gradients of chemical cues in solution. Haptotaxis, directional migration in response to gradients of substrate-bound cues, has received comparatively less attention; however, it is increasingly clear that in vivo many physiologically relevant guidance proteins – including many secreted cues – are bound to cellular surfaces or incorporated into extracellular matrix and likely function via a haptotactic mechanism. Here, we review the history of haptotaxis. We examine the importance of the reference surface, the surface in contact with the cell that is not covered by the cue, which forms a gradient opposing the gradient of the protein cue and must be considered in experimental designs and interpretation of results. We review and compare microfluidics, contact printing, light patterning, and 3D fabrication to pattern substrate-bound protein gradients in vitro. The range of methods to create substrate-bound gradients discussed herein makes possible systematic analyses of haptotactic mechanisms. Furthermore, understanding the fundamental mechanisms underlying cell motility will inform bioengineering approaches to program cell navigation and recover lost function.

show abstract

“…Recently, methods for creating concentration gradients of ions [7], small molecules [8], peptides [9], proteins [10], physiological parameters of the extracellular matrix [11,12] and cells [13] have been developed to construct research environments that mimic those found in vivo. To generate these gradients, diffusion [14], heat [15], gravity [16], electric force [17], light [18] and precise fluid control using microfluidic devices [12,[19][20][21][22] have been used. However, in most of these previous studies, the source of the biological compound was fixed in position and the concentration gradient remained constant over time with a predetermined spatial pattern.…”

Section: Introductionmentioning

confidence: 99%

Stepwise construction of dynamic microscale concentration gradients around hydrogel-encapsulated cells in a microfluidic perfusion culture device

et al. 2020

View full text Add to dashboard Cite

Inside living organisms, concentration gradients dynamically change over time as biological processes progress. Therefore, methods to construct dynamic microscale concentration gradients in a spatially controlled manner are needed to provide more realistic research environments. Here, we report a novel method for the construction of dynamic microscale concentration gradients in a stepwise manner around cells in micropatterned hydrogel. In our method, cells are encapsulated in a photodegradable hydrogel formed inside a microfluidic perfusion culture device, and perfusion microchannels are then fabricated in the hydrogel by micropatterned photodegradation. The cells in the micropatterned hydrogel can then be cultured by perfusing culture medium through the fabricated microchannels. By using this method, we demonstrate the simultaneous construction of two dynamic concentration gradients, which allowed us to expose the cells encapsulated in the hydrogel to a dynamic microenvironment.

show abstract

Independently Controlling Protein Dot Size and Spacing in Particle Lithography

Cited by 21 publications

References 46 publications

Ligand Nano-cluster Arrays in a Supported Lipid Bilayer

Ligand Nano-cluster Arrays in a Supported Lipid Bilayer

Substrate-Bound Protein Gradients to Study Haptotaxis

Stepwise construction of dynamic microscale concentration gradients around hydrogel-encapsulated cells in a microfluidic perfusion culture device

Contact Info

Product

Resources

About