High-throughput nitrogen-vacancy center imaging for nanodiamond photophysical characterization and pH nanosensing

Sow, Maabur; Steuer, Horst; Adekanye, Sanmi; Ginés, Laia; Mandal, Soumen; Gilboa, Barak; Williams, Oliver A.; Smith, Jason M.; Kapanidis, Achillefs N.

doi:10.1039/d0nr05931e

Cited by 25 publications

(24 citation statements)

References 57 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…When combined with pH-responsive polymers, their location can imply intracellular pH changes [ 19 ]. Charge conversion of NV centres can also be used as an indicator for pH but requires complex equipment for readout [ 20 ].…”

Section: Introductionmentioning

confidence: 99%

pH Sensitive Dextran Coated Fluorescent Nanodiamonds as a Biomarker for HeLa Cells Endocytic Pathway and Increased Cellular Uptake

Nie

Zhang

et al. 2021

Nanomaterials

View full text Add to dashboard Cite

Fluorescent nanodiamonds are a useful for biosensing of intracellular signaling networks or environmental changes (such as temperature, pH or free radical generation). HeLa cells are interesting to study with these nanodiamonds since they are a model cell system that is widely used to study cancer-related diseases. However, they only internalize low numbers of nanodiamond particles very slowly via the endocytosis pathway. In this work, we show that pH-sensitive, dextran-coated fluorescent nanodiamonds can be used to visualise this pathway. Additionally, this coating improved diamond uptake in HeLa cells by 5.3 times (*** p < 0.0001) and decreased the required time for uptake to only 30 min. We demonstrated further that nanodiamonds enter HeLa cells via endolysosomes and are eventually expelled by cells.

show abstract

Section: Introductionmentioning

confidence: 99%

pH Sensitive Dextran Coated Fluorescent Nanodiamonds as a Biomarker for HeLa Cells Endocytic Pathway and Increased Cellular Uptake

Nie

Zhang

et al. 2021

Nanomaterials

View full text Add to dashboard Cite

show abstract

“…To prepare the stock solutions of 10 mg/mL TiO 2 (5 and 60 nm; Nanostructured & Amorphous Materials, Katy, TX, USA), 10 mg/mL NDs (5-200 nm; Kay Diamond Products, Boca Raton, FL. USA) ( 56 , 57 ), and red fluorescent NDs (50 nm; brFND-50, nitrogen-vacancy NV centers per particle > 100, FND Biotech, Taipei, Taiwan) ( 58 , 59 ), the nanoparticles (NPs) were dispersed in distilled deionized water under sonication (80 W/L, 46 kHz) for 20 min. Test NP solutions were prepared immediately before use by dilution of the stock solutions with distilled deionized water and sonication (80 W/L, 46 kHz) for 20 min ( 60 ).…”

Section: Methodsmentioning

confidence: 99%

Nanodiamond-Induced Thrombocytopenia in Mice Involve P-Selectin-Dependent Nlrp3 Inflammasome-Mediated Platelet Aggregation, Pyroptosis and Apoptosis

Hung

Lien

et al. 2022

Front. Immunol.

View full text Add to dashboard Cite

Nanodiamond (ND) has been developed as a carrier to conduct various in vivo diagnostic and therapeutic uses. Safety is one of the major considerations, while the hemocompatibility of ND is not clearly addressed. Here we found that, compared to the other sizes of ND with relatively inert properties, treatments of 50 nm ND induced stronger platelet aggregation, platelet pyroptosis, apoptosis and thrombocytopenia in mice. Blockage treatments of soluble P-selectin, reactive oxygen species (ROS), and Nlrp3 inflammasome inhibitors markedly suppressed such adverse effects, suggesting ND-induced platelet activation and pyroptosis involves surface P-selectin-mediated enhancement of mitochondrial superoxide levels and Nlrp3 inflammasome activation. In addition, challenges of NDs induced less platelet pyroptosis and displayed less thrombocytopenia in P-selectin (Selp-/-), Nlrp3 (Nlrp3-/-) and caspase-1 (Casp1-/-) mutants, as compared to the wild type mice. Blockers of P-selectin, ROS, and Nlrp3 inflammasome pathways could be considered as antidotes for ND induced platelet activation and thrombocytopenia.

show abstract

“…The raw image was processed by home-built software (GapViewer) that detects diffraction-limited spots by their intensity (18). The software performs local background subtraction for each frame using the intensity around the Gaussian profile of the fluorescence emission from single DNA molecules.…”

Section: Methodsmentioning

confidence: 99%

“…Single-molecule imaging. Single-molecule fluorescence movies were collected using the Nanoimager-S single-molecule fluorescence microscope (Oxford Nanoimaging; (18,19)). The microscope was used as a widefield single-molecule fluorescence microscope with objective-based total internal reflection fluorescence (TIRF) illumination mode, with an excitation angle set at 52°.…”

Section: Preparation Of Labelled Cap Derivative (Alexa647-cap)mentioning

confidence: 99%

Transient DNA binding to gapped DNA substrates links DNA sequence to the single-molecule kinetics of protein-DNA interactions

Andrews

Steuer

El‐Sagheer

et al. 2022

Preprint

Self Cite

View full text Add to dashboard Cite

Protein interactions with nucleic acids are central to all genetic processes and many biotechnological applications. While many sequence-dependent protein-DNA interactions have been studied in detail using single-molecule methods, there is no standard high-throughput way to link the complex single-molecule kinetics of protein-DNA interactions with the DNA sequence of a single molecule. Here we provide the missing link by introducing a single-molecule imaging method (Gap-Seq) that interrogates DNA sequences via transient binding of short fluorescent DNA to a single DNA molecule previously used to characterise a protein-DNA interaction. In Gap-Seq, we identify a base by the degree of binding of 6-9 nt-long DNAs to surface-immobilised DNA substrates featuring a short single-stranded gap. To facilitate detection, we also developed a fluorescence quenching strategy that allows single-molecule detection at up to 500 nM of unbound fluorescent DNA. We link single-base differences on single DNA molecules to the kinetics of protein-DNA interactions by studying the interaction of a transcription activator with its cognate site. Finally, we show that our assay can address mixed sequences by distinguishing between two different sequences immobilised on the same field of view, paving the way for interrogation of sequence libraries for both mechanistic work and biotechnological applications.

show abstract

High-throughput nitrogen-vacancy center imaging for nanodiamond photophysical characterization and pH nanosensing

Abstract: A method to observe individual fluorescent crystal defects in nanodiamonds is reported and opens new nanosensing avenues (e.g. pH nanosensing).

Cited by 25 publications

References 57 publications

pH Sensitive Dextran Coated Fluorescent Nanodiamonds as a Biomarker for HeLa Cells Endocytic Pathway and Increased Cellular Uptake

pH Sensitive Dextran Coated Fluorescent Nanodiamonds as a Biomarker for HeLa Cells Endocytic Pathway and Increased Cellular Uptake

Nanodiamond-Induced Thrombocytopenia in Mice Involve P-Selectin-Dependent Nlrp3 Inflammasome-Mediated Platelet Aggregation, Pyroptosis and Apoptosis

Transient DNA binding to gapped DNA substrates links DNA sequence to the single-molecule kinetics of protein-DNA interactions

Contact Info

Product

Resources

About