Size-Tunable Organic Nanodot Arrays: A Versatile Platform for Manipulating and Imaging Cells

Pi, Fuwei; Dillard, Pierre; Alameddine, Ranime; Benard, Emmanuelle; Wahl, Astrid Klopstad; Ozerov, Igor; Charrier, Anne; Limozin, Laurent; Sengupta, Kheya

doi:10.1021/acs.nanolett.5b01400

Cited by 17 publications

(29 citation statements)

References 46 publications

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“…Major breakthroughs in SLB technologies include the ability to precisely pattern fluid and anchored ligands, to incorporate properly oriented and fluid transmembrane proteins, to generate multiple stacked bilayers, and to measure mechanical forces at the cell-SLB interface using ratiometric tension probes [25, 109, 110, 123, 136, 137]. Current studies of integrin and cadherin mediated adhesion using SLBs offer new insight into NA and AJ formation and demonstrate the power of spatiomechanical mutation using SLBs.…”

Section: Discussionmentioning

confidence: 99%

“…Thus, cells could engage with both laterally mobile and immobile ligands. In an alternate strategy, ligands were attached to size-tunable organic nanodot arrays (STONAs) surrounded by an SLB (Figure 3C) [123]. Beads deposited on a glass surface and coated in an aluminum mask of variable thickness determined STONA lattice spacing, ranging from 100 nm to 1800 nm.…”

Section: Supported Lipid Bilayers Technologiesmentioning

confidence: 99%

“…Scale bars 4 um. (Reprinted from [123] with permission from publisher). (D) Spatiomechanical mutation of receptor transport and phosphorylation on corralled bilayers.…”

Section: Figurementioning

confidence: 99%

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Supported lipid bilayer platforms to probe cell mechanobiology

Glazier¹,

Salaita²

2017

Biochimica et Biophysica Acta (BBA) - Biomembranes

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Mammalian and bacterial cells sense and exert mechanical forces through the process of mechanotransduction, which interconverts biochemical and physical signals. This is especially important in contact-dependent signaling, where ligand-receptor binding occurs at cell-cell or cell-ECM junctions. By virtue of occurring within these specialized junctions, receptors engaged in contact-dependent signaling undergo oligomerization and coupling with the cytoskeleton as part of their signaling mechanisms. While our ability to measure and map biochemical signaling within cell junctions has advanced over the past decades, physical cues remain difficult to map in space and time. Recently, supported lipid bilayer (SLB) technologies have emerged as a flexible platform to measure and manipulate membrane receptor mechanotransduction, allowing one to mimic cell-cell junctions. Changing the lipid composition and underlying substrate tunes bilayer fluidity, and lipid and ligand micro- and nano-patterning spatially control positioning and clustering of receptors. Patterning metal gridlines within SLBs introduces corrals that confine lipid mobility and introduce mechanical resistance. Here we review fundamental SLB mechanics and how SLBs can be engineered as tunable cell substrates for mechanotransduction studies. Finally, we highlight the impact of this work in understanding the biophysical mechanisms of cell adhesion.

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Section: Discussionmentioning

confidence: 99%

Section: Supported Lipid Bilayers Technologiesmentioning

confidence: 99%

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Supported lipid bilayer platforms to probe cell mechanobiology

Glazier¹,

Salaita²

2017

Biochimica et Biophysica Acta (BBA) - Biomembranes

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“…Colloidal bead lithography has been proposed as an alternative, and has been used either with gold chemistry 38 or with an all-organic approach 39 . The latter metal-free approach is compatible with TIRF and RICM imaging 39,40 , and is the technique of choice here. Our implementation has the additional advantage of offering variable pitch and dot-size.…”

Section: Introductionmentioning

confidence: 99%

Nano-clustering of ligands on surrogate antigen presenting cells modulates T cell membrane adhesion and organization

Dillard

Lellouch

et al. 2016

Integr. Biol.

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We investigate the adhesion and molecular organization of the plasma membrane of T lymphocytes interacting with a surrogate antigen presenting cell comprising glass supported ordered arrays of antibody (α-CD3) nano-dots dispersed in a non-adhesive matrix of polyethylene glycol (PEG). The local membrane adhesion and topography, as well as the distribution of the T cell receptors (TCRs) and the kinase ZAP-70, are influenced by dot-geometry, whereas the cell spreading area is determined by the overall average density of the ligands rather than specific characteristics of the dots. TCR clusters are recruited preferentially to the nano-dots and the TCR cluster size distribution has a weak dot-size dependence. On the patterns, the clusters are larger, more numerous, and more enriched in TCRs, as compared to the homogeneously distributed ligands at comparable concentrations. These observations support the idea that non-ligated TCRs residing in the non-adhered parts of the proximal membrane are able to diffuse and enrich the existing clusters at the ligand dots. However, long distance transport is impaired and cluster centralization in the form of a central supramolecular cluster (cSMAC) is not observed. Time-lapse imaging of early cell-surface contacts indicates that the ZAP-70 microclusters are directly recruited to the site of the antibody dots and this process is concomitant with membrane adhesion. These results together point to a complex interplay of adhesion, molecular organization and activation in response to spatially modulated stimulation.

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“…There has been also a constant effort to control more and more precisely the physical environment of the ligands in order to mimic one or more properties of cellular membranes and cellular material ( Figure 2A); this approach is often guided by the idea of mimicking as closely as possible the observed properties of APCs (Hivroz and Saitakis 2015). Metal barriers have been used to confine mobile ligands bound to a supported bilayer (Mossman 2005) mimicking corralling effects of the APC's cytoskeleton (Comrie 2015); ligands have been presented on islands of different spacing and size varying from micron (Doh and Irvine 2006), to sub-micron (Pi 2013(Pi , 2015 and nanometer (Delcassian 2013) scales. This helps to assess the role of different length scales on receptor behaviour and to reproduce various spatial organisations of receptors on the APC ( Figure 2B).…”

Section: Reductionist and Bio-mimetic Engineering Approachesmentioning

confidence: 99%

Membrane Organization and Physical Regulation of Lymphocyte Antigen Receptors: A Biophysicist’s Perspective

Limozin

Puech

2019

J Membrane Biol

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Receptors at the membrane of immune cells are the central players of innate and adaptative immunity, providing effective defence mechanisms against pathogens or cancer cells. Their function is intimately linked to their position at and within the membrane which provides accessibility, mobility as well as membrane proximal cytoskeleton anchoring, all of these elements playing important roles in the final function and links to cellular actions. Understanding how immune cells integrate the specific signals received at their membrane to take a decision remains an immense challenge and a very active field of fundamental and applied research. Recent progress in imaging and micromanipulation techniques have led to an unprecedented refinement in the description of molecular structures and supramolecular assemblies at the immune cell membrane, and provided a glimpse into their dynamics and regulation by force. Several key elements have been scrutinized such as the roles of geometry (relative sizes of molecules), lateral organisation, motion in the membrane of the receptors, but also physical cues such as forces, mediated by cellular substrates of different rigidities or applied by the cell itself, in conjunction with its partner cell. We review here these recent discoveries associated with a description of the biophysical methods used. While a conclusive picture integrating all of these components is still lacking, mainly due to the implication of diverse and different mechanisms and spatiotemporal scales involved, the amount of quantitative data available opens the way for physical modelling and numerical simulations and new avenues for experimental research.

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Size-Tunable Organic Nanodot Arrays: A Versatile Platform for Manipulating and Imaging Cells

Cited by 17 publications

References 46 publications

Supported lipid bilayer platforms to probe cell mechanobiology

Supported lipid bilayer platforms to probe cell mechanobiology

Nano-clustering of ligands on surrogate antigen presenting cells modulates T cell membrane adhesion and organization

Membrane Organization and Physical Regulation of Lymphocyte Antigen Receptors: A Biophysicist’s Perspective

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