Monolayer surface chemistry enables 2-colour single molecule localisation microscopy of adhesive ligands and adhesion proteins

Nicovich, Philip R.; Zhao, Manchen; Nieves, Daniel J.; Mollazade, Mahdie; Vivekchand, S. R. C.; Gaus, Katharina; Gooding, J. Justin

doi:10.1038/s41467-018-05837-7

Cited by 18 publications

(15 citation statements)

References 51 publications

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“…15a, b). We tested the functionality of RGD peptide by utilizing RGE peptide as a mutated control peptide 33 . We found that the subsequent formation of the Mg 2+ -BP NPs with RGE-BP did not significantly promote macrophage adhesion, when compared with the MgBNP dimer, thereby confirming the functionality of the RGD peptide to promote macrophage adhesion (Supplementary Fig.…”

Section: Resultsmentioning

confidence: 99%

Immunoregulation of macrophages by dynamic ligand presentation via ligand–cation coordination

et al. 2019

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Macrophages regulate host responses to implants through their dynamic adhesion, release, and activation. Herein, we employ bisphosphonate (BP)-coated gold nanoparticle template (BNP) to direct the swift and convertible formation of Mg 2+ -functional Mg 2+ -BP nanoparticle (NP) on the BP-AuNP surface via reversible Mg 2+ -BP coordination, thus producing (Mg 2+ -BP)-Au dimer (MgBNP). Ethylenediaminetetraacetic acid-based Mg 2+ chelation facilitates the dissolution of Mg 2+ -BP NP, thus enabling the reversion of the MgBNP to the BNP. This convertible nanoassembly incorporating cell-adhesive Mg 2+ moieties directs reversible attachment and detachment of macrophages by BP and EDTA, without physical scraping or trypsin that could damage cells. The swift formation of RGD ligand- and Mg 2+ -bifunctional RGD-Mg 2+ -BP NP that yields (RGD-Mg 2+ -BP)-Au dimer (RGDBNP) further stimulates the adhesion and pro-regenerative M2-type polarization of macrophages, both in vitro and in vivo, including rho-associated protein kinase. This swift and non-toxic dimer formation can include diverse bio-functional moieties to regulate host responses to implants.

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Section: Resultsmentioning

confidence: 99%

Immunoregulation of macrophages by dynamic ligand presentation via ligand–cation coordination

et al. 2019

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“…As such, the early forays into single molecule microscopy methods by biosensor researchers have mostly used these methods for characterizing how biointerfaces are working at a single molecule level. 25 The knowledge obtained using these sophisticated microscopies will suggest much simpler sensor technologies to assess specific molecular functions.…”

Section: T H Imentioning

confidence: 99%

Lighting Up Biosensors: Now and the Decade To Come

Ligler

Gooding

2019

Anal. Chem.

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Optical biosensors are defined as portable optical devices that use biorecognition molecules to interrogate a sample for the presence of a target. The capabilities of optical biosensors have expanded rapidly with advances in miniature optical components and molecular engineering. Biosensors to meet the needs in health and environmental monitoring and food safety have become commercially available, with many more in the pipeline. We review the innovative approaches to overcoming existing hurdles to practical biosensor designs and explore potential areas for future breakthroughs in optical biosensor technology.

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“…Secondly, surfaces that are both compatible with high‐resolution light microscopy and are electroactive are required. This typically means ITO, although studies of less common transparent materials are of interest . Of the molecules that have been shown to undergo electrochemical switching of their fluorescence, improvements are needed in photostability and fluorescence quantum yields .…”

Section: Figurementioning

confidence: 99%

Observing the Reversible Single Molecule Electrochemistry of Alexa Fluor 647 Dyes by Total Internal Reflection Fluorescence Microscopy

et al. 2019

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Alexa Fluor 647 is aw idely used fluorescent probe for cell bioimaging and super-resolution microscopy. Herein, the reversible fluorescence switching of Alexa Fluor 647 conjugated to bovine serum albumin (BSA) and adsorbed onto indium tin oxide (ITO) electrodes under electrochemical potential control at the level of single protein molecules is reported. The modulation of the fluorescence as af unction of potential was observed using total internal reflectance fluorescence (TIRF) microscopy. The fluorescence intensity of the Alexa Fluor 647 decreased, or reached background levels,a t reducing potentials but returned to normal levels at oxidizing potentials.T hese electrochemically induced changes in fluorescence were sensitive to pH despite that BSA-Alexa Fluor 647 fluorescence without applied potential is insensitive to pH between values of 4-10. The observed pH dependence indicated the involvement of electron and proton transfer in the fluorescence switching mechanism.

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Monolayer surface chemistry enables 2-colour single molecule localisation microscopy of adhesive ligands and adhesion proteins

Cited by 18 publications

References 51 publications

Immunoregulation of macrophages by dynamic ligand presentation via ligand–cation coordination

Immunoregulation of macrophages by dynamic ligand presentation via ligand–cation coordination

Lighting Up Biosensors: Now and the Decade To Come

Observing the Reversible Single Molecule Electrochemistry of Alexa Fluor 647 Dyes by Total Internal Reflection Fluorescence Microscopy

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