Developing Fluorescent Nanodiamonds for In Vitro and In Vivo Biological Imaging
At a Glance
Section titled âAt a Glanceâ| Metadata | Details |
|---|---|
| Publication Date | 2017-02-01 |
| Journal | Biophysical Journal |
| Authors | Keir C. Neuman |
| Institutions | National Institutes of Health |
| Citations | 1 |
| Analysis | Full AI Review Included |
Technical Documentation & Analysis: Advanced Diamond Materials for Bio-Imaging and Nanoscopy
Section titled âTechnical Documentation & Analysis: Advanced Diamond Materials for Bio-Imaging and NanoscopyâThis documentation analyzes the requirements outlined in the Biological Fluorescence Subgroup abstracts, focusing specifically on the critical role of high-ppurity, defect-engineered diamond materials in achieving next-generation in vitro and in vivo imaging capabilities.
Executive Summary
Section titled âExecutive SummaryâThe research abstracts highlight a critical need for highly stable, biocompatible fluorescent probes and ultra-flat substrates to advance single-molecule studies and super-resolution microscopy. 6CCVD is uniquely positioned to supply the necessary materials.
- Core Application: Development of Fluorescent Nanodiamonds (FNDs) utilizing the Nitrogen-Vacancy (NV-) center for superior biological imaging probes.
- Material Requirement: High-purity Single Crystal Diamond (SCD) is essential for the controlled creation and optimization of NV- centers, ensuring near-unity quantum efficiency and indefinite photo-stability.
- Key Achievement: FNDs enable high-resolution three-dimensional single-molecule imaging and in vivo background-free imaging via magnetic modulation.
- Microscopy Platform: Techniques like Atomic Force Microscopy (AFM) and Dual Resonance Frequency Enhanced Electrostatic Force Microscopy (DREEM) require ultra-flat, chemically inert diamond substrates (Ra < 1nm) for stable, high-precision measurements.
- Resolution Benchmark: RESOLFT nanoscopy variants achieved spatial resolution less than 50nm, demonstrating the demand for materials compatible with extreme optical precision.
- 6CCVD Value Proposition: We provide custom-sized, highly polished SCD wafers and plates, ideal for both NV center engineering and use as stable, low-noise microscopy platforms.
Technical Specifications
Section titled âTechnical SpecificationsâThe following data points extracted from the research define the performance metrics and material characteristics required for these advanced bio-imaging applications.
| Parameter | Value | Unit | Context |
|---|---|---|---|
| Spatial Resolution (RESOLFT) | < 50 | nm | Super-resolution imaging of rsFP-fusion proteins |
| NV Center Composition | N + V | Defect Pair | Negatively charged Nitrogen-Vacancy (NV-) center |
| NV Center Photo-Stability | Indefinite | - | No photo-bleaching or blinking observed |
| NV Center Quantum Efficiency | Near Unity | Ratio | Remarkable optical property for probes |
| DNA Replication Error Rate | 1 per 107 | bases | Uncorrected error frequency in replication |
| Imaging Speed Increase | 50 | -fold | Achieved using fast-switching RESOLFT variant |
| Substrate Flatness Requirement (AFM/DREEM) | Ra < 1 | nm | Required for high-precision single-molecule studies |
| Diamond Substrate Size (6CCVD Capability) | Up to 125 | mm | Custom dimensions for large microscopy stages |
Key Methodologies
Section titled âKey MethodologiesâThe research relies on sophisticated techniques that demand high-quality diamond materials for optimal performance, stability, and integration.
- Atomic Force Microscopy (AFM): Used for imaging mechanical properties of single biomolecules and live cells, and characterizing DNA repair complexes. Requires ultra-flat, rigid substrates.
- Dual Resonance Frequency Enhanced Electrostatic Force Microscopy (DREEM): A novel technique combining AFM topography with electrostatic potential mapping, requiring conductive or highly insulating, stable substrates.
- Single-Molecule Fluorescence Imaging (SMFI): Used to observe dynamics and reaction mechanisms of multi-protein complexes (e.g., DNA replication machinery).
- RESOLFT Optical Nanoscopy: A super-resolution technique that sequentially switches fluorescence capability on and off, achieving spatial resolution below 50nm.
- NV Center Magnetic Modulation: Utilizing the magnetic field-dependent fluorescence emission of NV- centers to achieve in vivo background-free imaging.
- Biocompatible Functionalization: Schemes developed for stabilizing and specifically labeling FNDs for biomedical applications.
6CCVD Solutions & Capabilities
Section titled â6CCVD Solutions & Capabilitiesâ6CCVD provides the foundational MPCVD diamond materials necessary to replicate, optimize, and scale the advanced bio-imaging and nanoscopy research described.
Applicable Materials for NV Center Engineering and Microscopy
Section titled âApplicable Materials for NV Center Engineering and Microscopyâ| Research Requirement | 6CCVD Material Solution | Technical Rationale |
|---|---|---|
| NV Center Precursor | Optical Grade Single Crystal Diamond (SCD) | High-purity, low-defect SCD is the ideal starting material for controlled nitrogen incorporation and subsequent vacancy creation (via irradiation/annealing) to maximize NV- yield and coherence time. |
| AFM/DREEM Substrates | High-Purity SCD Plates | SCD offers exceptional thermal stability, chemical inertness, and mechanical rigidity, crucial for minimizing drift and noise in nanoscale force measurements. |
| Conductive Substrates (DREEM) | Boron-Doped Diamond (BDD) | BDD provides tunable conductivity, essential for electrostatic potential mapping (DREEM) and integration of on-chip electrodes for localized field control. |
| Large-Area Stages | Polycrystalline Diamond (PCD) Wafers | For large-format microscopy stages or heat spreaders, 6CCVD offers PCD plates up to 125mm in diameter, providing superior thermal management and rigidity compared to traditional materials. |
Customization Potential for Advanced Research
Section titled âCustomization Potential for Advanced ResearchâThe complexity of single-molecule studies and nanoscopy often requires highly tailored material specifications. 6CCVD specializes in meeting these unique demands:
- Ultra-Low Roughness Polishing: We guarantee surface roughness (Ra) < 1nm on SCD and < 5nm on inch-size PCD, critical for minimizing background noise and ensuring stable protein adsorption in AFM/DREEM experiments.
- Custom Dimensions and Thickness: We supply SCD and PCD plates/wafers in custom dimensions up to 125mm, with thicknesses ranging from 0.1”m (for thin optical windows) up to 500”m (for rigid substrates) or 10mm (for bulk substrates).
- Integrated Metalization: For DREEM or magnetic modulation experiments requiring integrated electrodes, 6CCVD offers in-house metalization services, including deposition of Ti, Pt, Au, Pd, W, and Cu layers, patterned to customer specifications.
- Defect Engineering Support: Our material scientists can consult on optimizing nitrogen concentration and post-growth processing parameters (e.g., annealing temperature/time) to maximize the yield and quality of the NV- centers for FND production or bulk sensing applications.
Engineering Support
Section titled âEngineering Supportâ6CCVDâs in-house PhD team possesses deep expertise in CVD diamond growth and defect physics. We can assist researchers in material selection and specification development for similar NV Center Quantum Sensing and Super-Resolution Microscopy projects, ensuring the diamond substrate meets the stringent requirements for quantum coherence and optical clarity.
For custom specifications or material consultation, visit 6ccvd.com or contact our engineering team directly.