Real-Time Monitoring of the Evaporation and Fission of Electrospray-Ionized Polystyrene Beads and Bacterial Pellets at Elevated Temperatures.

This study uses real-time monitoring, at microsecond time scales, with a charge-sensing particle detector to investigate the evaporation and fission processes of methanol/micrometer-sized polystyrene beads (PS beads) droplets and bacterial particles droplets generated via electrospray ionization (ESI) under elevated temperatures. By incrementally raising capillary temperatures, the solvent, such as methanol on 0.75 µm PS beads, experiences partial evaporation. Further temperature increase induces fission, and methanol molecules continue to evaporate until PS ions are detected after this range. Similar partial evaporation is observed on 3 µm PS beads. However, the shorter period of the fission temperature range is necessary compared to 0.75 µm PS beads. For the spherical-shaped bacterium, Staphylococcus aureus, the desolvation process shows a similar fission period as compared to 0.75 µm PS beads. Comparably, the rod-shaped bacteria, Escherichia coli EC11303, and E. coli strain W have shorter fission periods than S. aureus. This research provides insights into the evaporation and fission mechanisms of ESI droplets containing different sizes and shapes of micrometer-sized particles, contributing to a better understanding of gaseous macroion formation.

Fluorescent nanodiamond immunosensors for clinical diagnostics of tuberculosis.

Fluorescent nanodiamonds (FNDs) are carbon nanoparticles containing a dense ensemble of nitrogen-vacancy defects as color centers. These centers have exceptional photostability and unique quantum properties, making them useful for ultrasensitive biosensing applications. This work employed FNDs conjugated with antibodies as magneto-optical immunosensors for tuberculosis (TB) diagnostics using competitive spin-enhanced lateral flow immunoassay (SELFIA). ESAT6 (6-kDa early secretory antigenic target) of Mycobacterium tuberculosis is a clinical marker of TB. We evaluated the assay's performance using the recombinant ESAT6 antigen and its antibodies noncovalently coated on FNDs. A detection limit of ∼0.02 ng mL-1 was achieved with the lateral flow membrane strip pre-structured with a narrow channel of 1 mm width. Adopting a cut-off value of 24.0 ng mm-1 for 100-nm FNDs on the strips, the method detected 49 out of 50 clinical samples with Mycobacterium tuberculosis complexes. In contrast, none of the assays for 10 clinical samples with non-tuberculous mycobacteria (NTM) isolates exhibited the presence of ESAT6. These results suggest that the SELFIA platform is applicable for TB detection and can differentiate TB from NTM infections, which also affect the human respiratory system. The FND-enabled immunosensing techniques are versatile and promising for early detection of TB and other diseases, opening a new avenue for biomedical applications of carbon-based nanomaterials.

Imaging Extreme Ultraviolet Radiation Using Nanodiamonds with Nitrogen-Vacancy Centers.

Extreme ultraviolet (EUV) radiation with wavelengths of 10-121 nm has drawn considerable attention recently for its use in photolithography to fabricate nanoelectronic chips. This study demonstrates, for the first time, fluorescent nanodiamonds (FNDs) with nitrogen-vacancy (NV) centers as scintillators to image and characterize EUV radiations. The FNDs employed are ∼100 nm in size; they form a uniform and stable thin film on an indium-tin-oxide-coated slide by electrospray deposition. The film is nonhygroscopic and photostable and can emit bright red fluorescence from NV0 centers when excited by EUV light. An FND-based imaging device has been developed and applied for beam diagnostics of 50 nm and 13.5 nm synchrotron radiations, achieving a spatial resolution of 30 µm using a film of ∼1 µm thickness. The noise equivalent power density is 29 µW/(cm2 Hz1/2) for the 13.5 nm radiation. The method is generally applicable to imaging EUV radiation from different sources.

Nanodiamonds-in-oil emulsions elicit potent immune responses for effective vaccination and therapeutics.

Background: The use of nanodiamonds (NDs) and fluorescent nanodiamonds (FNDs) as nonallergenic biocompatible additives in incomplete Freund's adjuvant (IFA) to elicit immune responses in vivo was investigated. Methods: C57BL/6 mice were immunized with chicken egg ovalbumin (OVA) in IFA and also OVA-conjugated NDs (or OVA-conjugated FNDs) in IFA to produce antibodies. OVA-expressing E.G7 lymphoma cells and OVA-negative EL4 cells were inoculated in mice to induce tumor formation. Results: The new formulation significantly enhanced immune responses and thus disease resistance. It exhibited specific therapeutic activities, effectively inhibiting the growth of E.G7 tumor cells in mice over 35 days. Conclusion: The high biocompatibility and multiple functionalities of NDs/FNDs render them applicable as active and trackable vaccine adjuvants and antitumor agents.

Antibody-Conjugated Nanodiamonds as Dual-Functional Immunosensors for In Vitro Diagnostics.

Nanodiamonds (NDs) are carbon nanoparticles with a large refractive index, a high density, and exceptional chemical stability. When excited by green light, they can emit bright red fluorescence from implanted nitrogen-vacancy (NV) centers. Taking advantage of these properties, we have developed antibody-conjugated NDs as in vitro diagnostic sensors for two complementary assays: particle-enhanced turbidimetric immunoassay (PETIA) and spin-enhanced lateral flow immunoassay (SELFIA). To achieve this goal, monocrystalline diamond powders (∼100 nm in diameter) with or without NV implantation were first treated in molten KNO3 to reduce their size and shape inhomogeneity, followed by surface carboxylation in strong oxidative acids and non-covalent conjugation with antibodies in water. PETIA and SELFIA were carried out separately with a microplate reader and a magnetically modulated fluorescence analyzer. Using C-reactive protein (CRP) as the target antigen, we found that anti-CRP-conjugated NDs exhibited high colloidal stability over 1 month at 4 °C in buffer solution. The limits of detection for 3 µL of CRP sample solution were 0.06 µg/mL and 1 ng/mL with variation coefficients of less than 10 and 15% for PETIA and SELFIA, respectively. These two methods together provide a detection range of 1 ng/mL-10 µg/mL, potentially useful for clinical applications. This work represents the first practical use of rounded monocrystalline NDs as in vitro diagnostic reagents.

Correlative Fluorescence and Transmission Electron Microscopy Assisted by 3D Machine Learning Reveals Thin Nanodiamonds Fluoresce Brighter.

Nitrogen vacancy (NV) centers in fluorescent nanodiamonds (FNDs) draw widespread attention as quantum sensors due to their room-temperature luminescence, exceptional photo- and chemical stability, and biocompatibility. For bioscience applications, NV centers in FNDs offer high-spatial-resolution capabilities that are unparalleled by other solid-state nanoparticle emitters. On the other hand, pursuits to further improve the optical properties of FNDs have reached a bottleneck, with intense debate in the literature over which of the many factors are most pertinent. Here, we describe how substantial progress can be achieved using a correlative transmission electron microscopy and photoluminescence (TEMPL) method that we have developed. TEMPL enables a precise correlative analysis of the fluorescence brightness, size, and shape of individual FND particles. Augmented with machine learning, TEMPL can be used to analyze a large, statistically meaningful number of particles. Our results reveal that FND fluorescence is strongly dependent on particle shape, specifically, that thin, flake-shaped particles are up to several times brighter and that fluorescence increases with decreasing particle sphericity. Our theoretical analysis shows that these observations are attributable to the constructive interference of light waves within the FNDs. Our findings have significant implications for state-of-the-art sensing applications, and they offer potential avenues for improving the sensitivity and resolution of quantum sensing devices.

Nanomaterial-based biosensors for avian influenza virus: A new way forward.

Avian influenza virus (AIV) is a zoonotic virus that can be transmitted from animals to humans. Although human infections are rare, the virus has a high mortality rate when contracted. Appropriate detection methods are thus crucial for combatting this pathogen. There is a growing demand for rapid, selective, and accurate methods of identifying the virus. Numerous biosensors have been designed and commercialized to detect AIV. However, they all have considerable shortcomings. Nanotechnology offers a new way forward. Nanomaterials produce more eco-friendly, rapid, and portable diagnostic systems. They also exhibit high sensitivity and selectivity while achieving a low detection limit (LOD). This paper reviews state-of-the-art nanomaterial-based biosensors for AIV detection, such as those composed of quantum dots, gold, silver, carbon, silica, nanodiamond, and other nanoparticles. It also offers insight into potential trial protocols for creating more effective methods of identifying AIV and discusses key issues associated with developing nanomaterial-based biosensors.

Variations in high density cholesterol levels based on apolipoprotein E variant and exercise type.

In various cross-sectional and longitudinal studies, exercise has been associated with cardiometabolic outcomes, including high-density lipoprotein (HDL) cholesterol. Exercise-induced changes in HDL cholesterol seem to be affected by genetic polymorphisms. In this study, we examined whether variant APOE rs7412 is involved in the association between HDL cholesterol and exercise. From adults assessed in Taiwan Biobank (TWB) between 2008 and 2019, we analyzed data from 57,638 normolipidemic subjects. To examine the association between exercise, APOE rs7412, and HDL cholesterol, a multiple linear regression model was used. A higher HDL was associated with both aerobic exercise (regression coefficient [mg/dL] beta- (ß), 1.112; 95% confidence interval (CI); 0.903-1.322) and resistance exercise (ß, 2.530; 95% CI, 2.093-2.966). In comparison with the APOE rs7412-CC genotype, the ß was 2.589 (95% CI, 2.329-2.848) among those with the CT + TT genotype. Compared to adults who had the CC genotype and did not exercise (the CC/no exercise group), the ß-coefficient determined for the different genotype and exercise groups was 1.135 (95% CI, 0.911-1.359) for the CC genotype and aerobic exercise group, 2.753 (95% CI, 2.283-3.322) for the CC genotype and resistance exercise group, 2.705 (95% CI, 2.390-3.020) for the CT + TT genotype and no exercise group, 3.682 (95% CI, 3.218-4.146) for the CT + TT genotype and aerobic exercise group, and 3.855 (95% CI, 2.727-4.982) for the CT + TT genotype and resistance exercise group, respectively. This study demonstrates that self-reported aerobic and resistance exercise both raised HDL levels, yet resistance exercise was associated with a greater increase, particularly among Taiwanese subjects carrying the APOE rs7412-CT+TT genotype.

A Novel Endodontic Approach in Removing Smear Layer Using Nano and Submicron Diamonds with Intracanal Oscillation Irrigation.

Sodium hypochlorite (NaOCl) and ethylenediaminetetraacetic acid (EDTA) are commonly recommended for effectively removing organic and inorganic components in the smear layer. This layer is found on root canal walls after root canal instrumentation. However, high-concentration EDTA reduces the strength of dentin and the dissolution efficacy of organic substances in NaOCl solution. The objective of this study was to investigate the efficacy of applying nano and submicron diamonds in irrigation solutions with sonic and ultrasonic oscillation for removing the smear layer during endodontic treatment. Extracted single-rooted human teeth were instrumented with ProTaper® Gold (Dentsply Sirona) nickel-titanium rotary instruments. Subsequently, each canal was irrigated with 3% NaOCl, 17% EDTA, distilled water, and 10-1000 nm-sized nano and submicron diamond irrigation solutions, respectively. Sonic and ultrasonic instruments were compared for oscillating the irrigation solutions. The teeth were processed for scanning electron microscopy to observe the efficiency of smear layer removal on the canal walls. Our results indicated that diamond sizes of 50 nm and above irrigation solutions showed significant effectiveness in removing the smear layer following the oscillation of sonic instruments for 10 s. Ultrasonic assisted 500 nm and 1000 nm diamond solutions significantly differed from the other diamond-sized solution in their ability to remove the smear layer. These results suggest that sonic and ultrasonic oscillation with specific sizes of nano and submicron diamond irrigation solution can be used as an alternative approach to removing the smear layer during endodontic treatment. The potential clinical application of root canal treatments can be expected.

Spin-Enhanced Lateral Flow Immunoassay for High-Sensitivity Detection of Nonstructural Protein NS1 Serotypes of the Dengue Virus.

Dengue fever is a global mosquito-borne viral infectious disease that has, in recent years, rapidly spread to almost all regions of the world. Lack of vaccination and directed treatment makes detection at the infection's early stages extremely important for disease prevention and clinical care. In this paper, we developed a rapid and highly sensitive dengue detection tool using a novel platform of diagnosis, called spin-enhanced lateral flow immunoassay (SELFIA) with a fluorescent nanodiamond (FND) as a reporter. Taking advantage of the unique magneto-optical properties of negatively charged nitrogen-vacancy centers in the FND, the SELFIA platform utilizes alternating electromagnetic fields to modulate signals from FND's fluorescence to provide sensitive and specific results. With sandwich SELFIA, we could efficiently detect all four dengue non-structural protein (NS1) serotypes (DV1, DV2, DV3, and DV4). The lowest detection concentration of the dengue NS1 antigens varied from 0.1 to 1.3 ng/mL, which is among the lowest limits of detection to date. The FND-based SELFIA technique is up to 500 and 5000 times more sensitive than carbon black and conventional gold nanoparticles, respectively. By using different anti-NS1 antibodies, we could differentiate the NS1 antigen serotypes contained in the tested samples via three simultaneous assays. Proposed SELFIA allows for both qualitative and quantitative differentiation between different NS1 protein serotypes, which will assist in the development of a highly sensitive and specific detection platform for dengue screening that has the potential to detect the disease at its early stages, especially in high-risk and limited-resource areas.

Photoluminescence of Nitrogen-Vacancy Centers by Ultraviolet One- and Two-Photon Excitation of Fluorescent Nanodiamonds.

Fluorescent nanodiamonds contain nitrogen-vacancy (NV) centers as quantum defects. When exposed to a continuous-wave 325 nm laser or a femtosecond 344 nm laser, the particles emit red fluorescence from NV0 centers at ∼620 nm. Power dependence measurements of the emission strength revealed a predominantly linear behavior at the laser peak intensity lower than 1 GW·cm-2, contributed mainly by photoexcitation of electrons from the valence band of diamond to the NV0 centers, followed by relaxation via electron-hole recombination. In the higher power regions, however, nonresonant two-photon interband excitation of the diamond matrix dominates the photoluminescence processes. Best fits of the experimental data to semiempirical models revealed an ionization coefficient of ∼1 cm-1 for the one-photon valence-to-defect excitation and a saturation intensity of 180 ± 60 GW·cm-2 for the two-photon interband excitation. The study provides new insight into the photoionization of NV0 centers and the interband excitation properties of diamond in the UV region.

Fluorescent nanodiamond-based spin-enhanced lateral flow immunoassay for detection of SARS-CoV-2 nucleocapsid protein and spike protein from different variants.

SARS-CoV-2 viruses, responsible for the COVID-19 pandemic, continues to evolve into new mutations, which poses a significant threat to public health. Current testing methods have some limitations, such as long turnaround times, high costs, and professional laboratory requirements. In this report, the novel Spin-Enhanced Lateral Flow Immunoassay (SELFIA) platform and fluorescent nanodiamond (FND) reporter were utilized for the rapid detection of SARS-CoV-2 nucleocapsid and spike antigens from different variants, including wild-type (Wuhan-1), Alpha (B.1.1.7), Delta (B.1.617.2), and Omicron (B.1.1.529). The SARS-CoV-2 antibodies were conjugated with FND via nonspecific binding, enabling the detection of SARS-CoV-2 antigens via both direct and competitive SELFIA format. Direct SELFIA was performed by directly adding the SARS-CoV-2 antibodies-conjugated FND on the antigens-immobilized nitrocellulose (NC) membrane. Conversely, the SARS-CoV-2 antigen-containing sample was first incubated with the antibodies-conjugated FND, and then dropped on the antigen-immobilized NC membrane to carry out the competitive SELFIA. The results suggested that S44F anti-S IgG antibody can be efficiently used for the detection of wild-type, Alpha, Delta, and Omicron variants spike antigens. Findings were comparable in direct SELFIA, competitive SELFIA, and ELISA. A detection limit of 1.94, 0.77, 1.14, 1.91, and 1.68 ng/mL can be achieved for SARS-CoV-2 N protein, wild-type, Alpha, Delta, and Omicron S proteins, respectively, via competitive SELFIA assay. These results suggest that a direct SELFIA assay can be used for antibody/antigen pair screening in diagnosis development, while the competitive SELFIA assay can serve as an accurate quantitative diagnostic tool. The simplicity and rapidity of the SELFIA platform were demonstrated, which can be leveraged in the detection of other infectious diseases in the near future.

Retention in Community Health Screening among Taiwanese Adults: A 9-Year Prospective Cohort Study.

Largely conducted in Western developed nations, research on community health screening has mainly been of limited duration. This study aims to ascertain the predictors of retention in a community health screening program, involving multiple admission cohorts over a 9-year period in Taiwan. Retention is defined as the participation in subsequent waves of health screening after being recruited for an initial screening. Data came from a prospective cohort study, named "Landseed Integrated Outreaching Neighborhood Screening (LIONS)", in Taiwan. This research retrieved 5901 community-dwelling Taiwanese adults aged 30 and over from LIONS and examined their retention in three follow-ups during 2006-2014. Generalized estimating equations were employed to evaluate retention over time as a function of social determinants, health behaviors, and health conditions. Being middle-aged, higher education, and regular exercise were positively associated with retention. Conversely, smoking, betel-nut chewing, psychiatric disorder, hypertension, type 2 diabetes mellitus, stroke, and a longer time interval since enrollment were negatively associated with retention. Furthermore, retention rates varied substantially across admission cohorts with more recent cohorts having a lower rate of retention (aOR = 0.33-0.83). Greater attention needs to be directed to retention over time and variations across admission cohorts. Additionally, those who are in either younger or older age groups and have chronic diseases or unhealthy behaviors should be targeted with greater efforts.

Thermometric lateral flow immunoassay with colored latex beads as reporters for COVID-19 testing.

Temperature sensing is a promising method of enhancing the detection sensitivity of lateral flow immunoassay (LFIA) for point-of-care testing. A temperature increase of more than 100 °C can be readily achieved by photoexcitation of reporters like gold nanoparticles (GNPs) or colored latex beads (CLBs) on LFIA strips with a laser power below 100 mW. Despite its promise, processes involved in the photothermal detection have not yet been well-characterized. Here, we provide a fundamental understanding of this thermometric assay using non-fluorescent CLBs as the reporters deposited on nitrocellulose membrane. From a measurement for the dependence of temperature rises on the number density of membrane-bound CLBs, we found a 1.3-fold (and 3.2-fold) enhancement of the light absorption by red (and black) latex beads at 520 nm. The enhancement was attributed to the multiple scattering of light in this highly porous medium, a mechanism that could make a significant impact on the sensitivity improvement of LFIA. The limit of detection was measured to be 1 × 105 particles/mm2. In line with previous studies using GNPs as the reporters, the CLB-based thermometric assay provides a 10× higher sensitivity than color visualization. We demonstrated a practical use of this thermometric immunoassay with rapid antigen tests for COVID-19.

Recent Advances in Novel Lateral Flow Technologies for Detection of COVID-19.

The development of reliable and robust diagnostic tests is one of the most efficient methods to limit the spread of coronavirus disease 2019 (COVID-19), which is caused by the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). However, most laboratory diagnostics for COVID-19, such as enzyme-linked immunosorbent assay (ELISA) and reverse transcriptase-polymerase chain reaction (RT-PCR), are expensive, time-consuming, and require highly trained professional operators. On the other hand, the lateral flow immunoassay (LFIA) is a simpler, cheaper device that can be operated by unskilled personnel easily. Unfortunately, the current technique has some limitations, mainly inaccuracy in detection. This review article aims to highlight recent advances in novel lateral flow technologies for detecting SARS-CoV-2 as well as innovative approaches to achieve highly sensitive and specific point-of-care testing. Lastly, we discuss future perspectives on how smartphones and Artificial Intelligence (AI) can be integrated to revolutionize disease detection as well as disease control and surveillance.

Relaxation of a dense ensemble of spins in diamond under a continuous microwave driving field.

Decoherence of Rabi oscillation in a two-level quantum system consists of two components, a simple exponential decay and a damped oscillation. In dense-ensemble spin systems like negatively charged nitrogen-vacancy (NV-) centers in diamond, fast quantum state decoherence often obscures clear observation of the Rabi nutation. On the other hand, the simple exponential decay (or baseline decay) of the oscillation in such spin systems can be readily detected but has not been thoroughly explored in the past. This study investigates in depth the baseline decay of dense spin ensembles in diamond under continuously driving microwave (MW). It is found that the baseline decay times of NV- spins decrease with the increasing MW field strength and the MW detuning dependence of the decay times shows a Lorentzian-like spectrum. The experimental findings are in good agreement with simulations based on the Bloch formalism for a simple two-level system in the low MW power region after taking into account the effect of inhomogeneous broadening. This combined investigation provides new insight into fundamental spin relaxation processes under continuous driving electromagnetic fields and paves ways to better understanding of this underexplored phenomena using single NV- centers, which have shown promising applications in quantum computing and quantum metrology.

Effects of Size and Surface Properties of Nanodiamonds on the Immunogenicity of Plant-Based H5 Protein of A/H5N1 Virus in Mice.

Nanodiamond (ND) has recently emerged as a potential nanomaterial for nanovaccine development. Here, a plant-based haemagglutinin protein (H5.c2) of A/H5N1 virus was conjugated with detonation NDs (DND) of 3.7 nm in diameter (ND4), and high-pressure and high-temperature (HPHT) oxidative NDs of ~40-70 nm (ND40) and ~100-250 nm (ND100) in diameter. Our results revealed that the surface charge, but not the size of NDs, is crucial to the protein conjugation, as well as the in vitro and in vivo behaviors of H5.c2:ND conjugates. Positively charged ND4 does not effectively form stable conjugates with H5.c2, and has no impact on the immunogenicity of the protein both in vitro and in vivo. In contrast, the negatively oxidized NDs (ND40 and ND100) are excellent protein antigen carriers. When compared to free H5.c2, H5.c2:ND40, and H5.c2:ND100 conjugates are highly immunogenic with hemagglutination titers that are both 16 times higher than that of the free H5.c2 protein. Notably, H5.c2:ND40 and H5.c2:ND100 conjugates induce over 3-folds stronger production of both H5.c2-specific-IgG and neutralizing antibodies against A/H5N1 than free H5.c2 in mice. These findings support the innovative strategy of using negatively oxidized ND particles as novel antigen carriers for vaccine development, while also highlighting the importance of particle characterization before use.

Magnetically Modulated Fluorescence of Nitrogen-Vacancy Centers in Nanodiamonds for Ultrasensitive Biomedical Analysis.

The negatively charged nitrogen-vacancy center in fluorescent nanodiamonds (FNDs) is a point defect with unique magneto-optical properties. It emits far-red fluorescence at ∼700 nm, and its intensity can be magnetically modulated with a depth of more than 10% at a field strength of 30 mT. We have closely examined this property and illustrated its practical use in biomedicine by applying a periodic, time-varying magnetic field to FNDs deposited on a surface or dispersed in a solution with a lock-in detection method. We achieved selective and sensitive detection of 100 nm FNDs on a nitrocellulose membrane at a particle density of 0.04 ng/mm2 (or ∼2 × 104 particles/mm2) and in an aqueous solution with a particle concentration of 1 ng/mL (or ∼1 fM) in 10 s as the detection limits. The utility and versatility of the technique were demonstrated with an application to background-free detection of FNDs as reporters for FND-based lateral flow immunoassays as well as selective quantification of FNDs in tissue digests for in vivo studies.

Mesenchymal stem/stromal cell-based therapy: mechanism, systemic safety and biodistribution for precision clinical applications.

Mesenchymal stem/stromal cells (MSCs) are a promising resource for cell-based therapy because of their high immunomodulation ability, tropism towards inflamed and injured tissues, and their easy access and isolation. Currently, there are more than 1200 registered MSC clinical trials globally. However, a lack of standardized methods to characterize cell safety, efficacy, and biodistribution dramatically hinders the progress of MSC utility in clinical practice. In this review, we summarize the current state of MSC-based cell therapy, focusing on the systemic safety and biodistribution of MSCs. MSC-associated risks of tumor initiation and promotion and the underlying mechanisms of these risks are discussed. In addition, MSC biodistribution methodology and the pharmacokinetics and pharmacodynamics of cell therapies are addressed. Better understanding of the systemic safety and biodistribution of MSCs will facilitate future clinical applications of precision medicine using stem cells.

Cell Volume (3D) Correlative Microscopy Facilitated by Intracellular Fluorescent Nanodiamonds as Multi-Modal Probes.

Three-dimensional correlative light and electron microscopy (3D CLEM) is attaining popularity as a potential technique to explore the functional aspects of a cell together with high-resolution ultrastructural details across the cell volume. To perform such a 3D CLEM experiment, there is an imperative requirement for multi-modal probes that are both fluorescent and electron-dense. These multi-modal probes will serve as landmarks in matching up the large full cell volume datasets acquired by different imaging modalities. Fluorescent nanodiamonds (FNDs) are a unique nanosized, fluorescent, and electron-dense material from the nanocarbon family. We hereby propose a novel and straightforward method for executing 3D CLEM using FNDs as multi-modal landmarks. We demonstrate that FND is biocompatible and is easily identified both in living cell fluorescence imaging and in serial block-face scanning electron microscopy (SB-EM). We illustrate the method by registering multi-modal datasets.