Rapid SERS Quantification of Trace Fentanyl Laced in Recreational Drugs with a Portable Raman Module.

Rapid identification and quantification of opioid drugs are of significant importance and an urgent need in drug regulation and control, considering the serious social and economic impact of the opioid epidemic in the United States. Unfortunately, techniques for accurate detection of these opioids, particularly for fentanyl, an extremely potent synthetic drug of abuse and a main perpetrator in the opioid crisis, are often not readily accessible. Therefore, a fast, highly sensitive, and preferably quantitative technique, with excellent portability, is highly desirable. Such a technique can potentially offer timely and crucial information for drug control officials, as well as health professionals, about drug distribution and overdose prevention. We therefore propose a portable surface-enhanced Raman scattering (SERS) approach by pairing an easy to perform yet reliable SERS protocol with a compact Raman module suitable for rapid, on-site identification and quantification of trace fentanyl. Fentanyl spiked in urine control was successfully detected at concentrations as low as 5 ng/mL. Portable SERS also enabled detection of trace fentanyl laced in recreational drugs at mass concentrations as low as 0.05% (5 ng in 10 µg total) and 0.1% (10 ng in 10 µg total) in heroin and tetrahydrocannabinol (THC), respectively. Drug interaction with the nanoparticle surface was simulated through molecular dynamics to investigate the molecular adsorption mechanism and account for SERS signal differences observed for opioid drugs. Furthermore, resolution of fentanyl in binary and ternary opioid mixtures was readily achieved with multivariate data analysis. In sum, we developed a rapid, highly sensitive, and reliably quantitative method for trace fentanyl analysis by synergizing a streamlined SERS procedure and a portable Raman module at low cost.

Quantifying and optimizing photocurrent via optical modeling of gold nanostar-, nanorod-, and dimer-decorated MoS2 and MoTe2.

Finite element simulations through COMSOL Multiphysics were used to optically model systems composed of Mo dichalcogenide layers (MoTe2 and MoS2) and Au nanoparticles (spherical dimers, nanorods, and nanostars) to understand how their fundamental material properties as well as their interactions affect the photocurrent response. The absorption cross sections of the various Au nanoparticles linearly increase with respect to their increasing dimensions, hence being ideal tunable systems for the enhancement of the electric field in the dichalcogenide layers under visible and near infrared. The photocurrent through the MoTe2 and MoS2 substrates was enhanced by the addition of Au nanoparticles when the plasmonic response was localized in the area of the particle in contact with the substrate. Based on these findings, the use of Au nanoparticles can greatly improve the unique photocurrent properties of Mo dichalcogenides; however, nanoparticle orientation and size must be considered to tune the enhancement at the specific wavelengths. This computational work provides useful design rules for the use of plasmonic nanomaterials in photocatalytic and photocurrent enhancement of transition metal dichalcogenides.

Nanopolystyrene translocation and fetal deposition after acute lung exposure during late-stage pregnancy.

BACKGROUND: Plastic is everywhere. It is used in food packaging, storage containers, electronics, furniture, clothing, and common single-use disposable items. Microplastic and nanoplastic particulates are formed from bulk fragmentation and disintegration of plastic pollution. Plastic particulates have recently been detected in indoor air and remote atmospheric fallout. Due to their small size, microplastic and nanoplastic particulate in the atmosphere can be inhaled and may pose a risk for human health, specifically in susceptible populations. When inhaled, nanosized particles have been shown to translocate across pulmonary cell barriers to secondary organs, including the placenta. However, the potential for maternal-to-fetal translocation of nanosized-plastic particles and the impact of nanoplastic deposition or accumulation on fetal health remain unknown. In this study we investigated whether nanopolystyrene particles can cross the placental barrier and deposit in fetal tissues after maternal pulmonary exposure. RESULTS: Pregnant Sprague Dawley rats were exposed to 20 nm rhodamine-labeled nanopolystyrene beads (2.64 × 1014 particles) via intratracheal instillation on gestational day (GD) 19. Twenty-four hours later on GD 20, maternal and fetal tissues were evaluated using fluorescent optical imaging. Fetal tissues were fixed for particle visualization with hyperspectral microscopy. Using isolated placental perfusion, a known concentration of nanopolystyrene was injected into the uterine artery. Maternal and fetal effluents were collected for 180 min and assessed for polystyrene particle concentration. Twenty-four hours after maternal exposure, fetal and placental weights were significantly lower (7 and 8%, respectively) compared with controls. Nanopolystyrene particles were detected in the maternal lung, heart, and spleen. Polystyrene nanoparticles were also observed in the placenta, fetal liver, lungs, heart, kidney, and brain suggesting maternal lung-to-fetal tissue nanoparticle translocation in late stage pregnancy. CONCLUSION: These studies confirm that maternal pulmonary exposure to nanopolystyrene results in the translocation of plastic particles to placental and fetal tissues and renders the fetoplacental unit vulnerable to adverse effects. These data are vital to the understanding of plastic particulate toxicology and the developmental origins of health and disease.

Improved Precision in Surface-Enhanced Raman Scattering Quantification of Analyte through Dual-Modality Multisite Sensing.

Accurate quantification of analyte using surface-enhanced Raman spectroscopy (SERS) is a desired, yet unfulfilled, ability that could enable a plethora of diagnostic- and defense-related applications. The major hurdles to overcome to achieve this goal are expensive manufacturing for highly ordered and reproducible substrates and low reproducibility of substrates produced through low cost methods. A technology that can set industry standards for manufacturing/processing of SERS substrates is still yet to be achieved. A dual-modality multisite sensing approach was developed, that overcomes the limitations experienced when fabricating bottom-up, reproducible, sensitive, and low-cost SERS substrates. Electrochemistry was combined with SERS for dual-modality sensing to improve precision by adding redundancy and encoding features, thus increasing measurement robustness and predictability. This technique works by calibrating SERS response with respect to active surface area, a parameter known to be proportional to charge, which can be estimated via electrochemical measurements. The dual-modality multisite measurement demonstrates at least 2.8× improvement in assay precision compared to the traditional single-site Raman measurements. The technique yields overall improved precision of measurement and is not limited to any particular SERS substrate or geometry, and thus it can be adapted and incorporated readily in any SERS sensing assay.

Impact of Protein Corona in Nanoflare-Based Biomolecular Detection and Quantification.

Selective detection and precise quantification of biomolecules in intracellular settings play a pivotal role in the diagnostics and therapeutics of diseases, including various cancers and infectious epidemics. Because of this clinical relevance, nanoprobes with high sensitivity, wide tunability, and excellent biological stability have become of high demand. In particular, nanoflares based on gold nanoparticles have emerged as an attractive candidate for intracellular detection due to their efficient cellular uptake, enhanced binding affinity with complementary targets, and improved biological compatibility. However, nanoprobes, including these nanoflares, are known to be susceptible to the adsorption of proteins present in the biological environment, which leads to the formation of a so-called protein corona layer on their surface, leading to an altered targeting efficiency and cellular uptake. In this work, we leverage the nanoflares platform to demonstrate the effect of protein corona on biomolecular detection, quantification, as well as biological stability against enzymatic degradation. Nanoflares incubated in a biologically relevant concentration of serum albumin proteins (0.50 wt %) were shown to result in more than 20% signal reduction in target detection, with a decrease varying proportionally with the protein concentrations. In addition, similar signal reduction was observed for different serum proteins, and PEG backfilling was found to be ineffective in mitigating the negative impact induced by the corona formation. Furthermore, nuclease resistance in nanoflares was also severely compromised by the presence of the corona shell (∼2-fold increase in hydrolysis activity). This work demonstrates the consequences of an in situ formed protein corona layer on molecular detection/quantification and biological stability of nanoflares in the presence of nuclease enzymes, highlighting the importance of calibrating similar nanoprobes in proper biological media to improve the accuracy of molecular detection and quantification.

Enhancing hot electron generation and injection in the near infrared via rational design and controlled synthesis of TiO2-gold nanostructures.

Plasmonic nanostructure/semiconductor composites are receiving great interest as powerful photocatalytic platforms able to increase solar energy conversion efficiency compared to more traditional approaches. The possibility to grow a thin titania shell onto the gold nanoparticle, thus substantially increasing the metal-semiconductor area of contact, is expected to be ideal for photocatalytic water reduction, especially if the titania (TiO2) coating displays limited thickness and high crystallinity. We argue however that the morphology of the underlying gold nanoparticle and the quality of the interface are the main drivers of photocatalytic performance. Herein, we show how we can synthesize TiO2-coated gold nanostar- and gold nanorod-based photocatalysts and identify the most important design parameters that one should be focusing on for the optimization of hot electron-based photocatalysts. In addition to nanoparticle morphology and interface quality, we determine that the integrated absorptivity of the plasmon band and the uniformity and crystallinity of the semiconductor shell are important, even though to a lesser extent. These results may prove interesting not only to increase production rates in hydrogen evolution reactions or other chemical conversions, but also to decouple and understand additional mechanisms driving photocatalysis, other than the sequential, hot electron mediated one, as we reported before.

SERS-Based Quantification of Biomarker Expression at the Single Cell Level Enabled by Gold Nanostars and Truncated Aptamers.

Surface-enhanced Raman spectroscopy (SERS)-based biosensors have been used increasingly over the past few years for cancer detection and diagnosis. SERS-based imaging offers excellent sensitivity and has advantages over other detection techniques such as fluorescence. In this study, we developed a novel biosensor to detect the cancer biomarker epithelial cell adhesion molecule (EpCAM) and quantify its expression at the single cell level. EpCAM is one of the most commonly expressed markers on a variety of cancer cells; importantly it has been suggested that reduction of its expression levels could be associated with the epithelial to mesenchymal transition (EMT) and thus to the onset of metastasis. Therefore, monitoring variations in expression levels of this membrane biomarker would improve our ability to monitor cancer progression. The described substrate-based biosensor was developed employing gold nanostars functionalized with EpCAM aptamer molecules and was able to quantify subnanomolar concentrations of EpCAM protein in solution. Importantly, we demonstrated its use to quantify EpCAM expression on the surface of two cancer cells, MCF-7 and PC-3. We also compared the binding efficiency of two EpCAM DNA aptamers of different lengths and observed a substantial improvement in the sensitivity of detection by employing the shorter aptamer sequence, probably due to the reduced number of conformations possible at room temperature with the truncated oligonucleotide. Detailed characterization of the substrates was carried out using both SERS maps and atomic force microscopy. These substrate-based diagnostic devices promise to be relevant for monitoring phenotype evolutions in cancer cells, blood, and other bodily fluids, thus improving our ability to follow in real time disease onset and progression.

Multiparametric Assessment of Gold Nanoparticle Cytotoxicity in Cancerous and Healthy Cells: The Role of Size, Shape, and Surface Chemistry.

In recent years, we and others have become interested in evaluating the use of surface-enhanced Raman scattering (SERS) tags for early cancer detection and in designing new approaches to demonstrate the applicability of this spectroscopic technique in the clinic. SERS-based imaging in particular offers ultra sensitivity up to the single molecule, multiplexing capability, and increased photostability and has been shown to outperform fluorescence. However, to employ SERS tags for early cancer detection, it is important to understand their interaction with cells and determine their cytotoxicity. We have been particularly interested for quite some time in determining if and how gold nanostars, which have been demonstrated as outstanding SERS enhancing substrates, can be safely employed in living systems and translated to the clinic. In this study, we carried out a multiparametric in vitro study to look at the cytotoxicity and cellular uptake of gold nanoparticles on human glioblastoma and human dermal fibroblast cell lines. Cytotoxicity was evaluated by incubating cells with three different morphologies of AuNPs, namely nanospheres, nanorods, and nanostars, each having three different surface chemistries (cetyltrimethylammonium bromide (CTAB), poly(ethylene glycol) (PEG), and human serum albumin (HSA)). Our results showed that the surface chemistry of the nanoparticles had predominant effects on cytotoxicity, and the morphology and size of the nanoparticles only slightly affected cell viability. CTAB-coated particles were found to be the most toxic to cells, and PEGylated nanostars were determined to be the least toxic. Caspase-3 assay and LDH assay revealed that cell death occurs via apoptosis for cancerous cells and via necrosis for healthy ones. Cellular uptake studies carried out via TEM showed that the particles retain their shape even at long incubation times, which may be beneficial for in vivo SERS-based disease detection. Overall, this study provides valuable information on gold-nanoparticle-induced cytotoxicity that can be leveraged for the development of safe and effective nanoparticle-based therapeutic and diagnostic systems.

Plasmonic properties of regiospecific core-satellite assemblies of gold nanostars and nanospheres.

Solution-based molecularly-mediated bottom-up assembly of gold nanostars and nanospheres in regiospecific core-satellite nanoarchitectures is reported. The controlled assembly is driven by coupling reactions in solution between small, rigid, Raman-active organic molecules bound to the surface of the nanoparticles, and leads to much narrower interparticle gaps than achievable with DNA-based assembly methods. In the described system, gold nanostars with multiple sharp spikes, ideal for electromagnetic field enhancement, are used as the core particle onto which spherical satellites are assembled. Transmission electron micrographs show that the core-satellite structures assemble with <2 nm interparticle gaps and regiospecific binding of only one sphere per spike, and the process can be followed by monitoring changes in the surface enhanced Raman scattering (SERS) spectra of the Raman active linkers. The assembled structures give rise on average to two orders of magnitude SERS signal enhancement per nanoparticle in comparison to their constituents, which can be attributed to the creation of SERS "hot spots" between the nanostar tip and the satellite sphere. Two dimensional finite element electromagnetic models show strongly confined electromagnetic field intensity in the narrow interparticle gaps of core-satellite assemblies, which is significantly enhanced in comparison to the constituent nanoparticles, thus corroborating the experimental findings. Thus, the assemblies reported here can be envisioned as SERS-tags for imaging purposes as well as a model system for SERS-based chemical sensing with improved sensitivity.

Discrimination of Genetic Biomarkers of Disease through Machine-Learning-Based Hypothesis Testing of Direct SERS Spectra of DNA and RNA.

Cancer is globally a leading cause of death that would benefit from diagnostic approaches detecting it in its early stages. However, despite much research and investment, cancer early diagnosis is still underdeveloped. Owing to its high sensitivity, surface-enhanced Raman spectroscopy (SERS)-based detection of biomarkers has attracted growing interest in this area. Oligonucleotides are an important type of genetic biomarkers as their alterations can be linked to the disease prior to symptom onset. We propose a machine-learning (ML)-enabled framework to analyze complex direct SERS spectra of short, single-stranded DNA and RNA targets to identify relevant mutations occurring in genetic biomarkers, which are key disease indicators. First, by employing ad hoc-synthesized colloidal silver nanoparticles as SERS substrates, we analyze single-base mutations in ssDNA and RNA sequences using a direct SERS-sensing approach. Then, an ML-based hypothesis test is proposed to identify these changes and differentiate the mutated sequences from the corresponding native ones. Rooted in "functional data analysis," this ML approach fully leverages the rich information and dependencies within SERS spectral data for improved modeling and detection capability. Tested on a large set of DNA and RNA SERS data, including from miR-21 (a known cancer miRNA biomarker), our approach is shown to accurately differentiate SERS spectra obtained from different oligonucleotides, outperforming various data-driven methods across several performance metrics, including accuracy, sensitivity, specificity, and F1-scores. Hence, this work represents a step forward in the development of the combined use of SERS and ML as effective methods for disease diagnosis with real applicability in the clinic.

Long term follow-up of heart rate variability in healthcare workers with mild COVID-19.

Introduction: Prior investigations into post-COVID dysautonomia often lacked control groups or compared affected individuals solely to healthy volunteers. In addition, no data on the follow-up of patients with SARS-CoV-2-related autonomic imbalance are available. Methods: In this study, we conducted a comprehensive clinical and functional follow-up on healthcare workers (HCWs) with former mild COVID-19 (group 1, n = 67), to delineate the trajectory of post-acute autonomic imbalance, we previously detected in a case-control study. Additionally, we assessed HCWs for which a test before SARS-CoV-2 infection was available (group 2, n = 29), who later contracted SARS-CoV-2, aiming to validate findings from our prior case-control investigation. We evaluated autonomic nervous system heart modulation by means of time and frequency domain heart rate variability analysis (HRV) in HCWs during health surveillance visits. Short-term electrocardiogram (ECG) recordings, were obtained at about 6, 13 months and both at 6 and 13 months from the negative SARS-CoV-2 naso-pharyngeal swab (NPS) for group 1 and at about 1-month from the negative NPS for group 2. HCWs who used drugs, had comorbidities that affected HRV, or were hospitalized with severe COVID-19 were excluded. Results: Group 1 was split into three subgroups clinically and functionally followed at, about 6 months (subgroup-A, n = 17), 13 months (subgroup-B, n = 37) and both at 6 and 13 months (subgroup-C, n = 13) from the negative SARS-CoV-2 NPS. In subgroup-A, at 6-month follow-up compared with baseline, the spectral components in the frequency domain HRV parameters, showed an increase in normalized high frequency power (nHF) (t = 2.99, p = 0.009), a decrease in the normalized low frequency power (nLF) (t = 2.98, p = 0.009) and in the LF/HF ratio (t = 3.13, p = 0.006). In subgroup B, the comparison of the spectral components in the frequency domain HRV parameters, at 13-month follow-up compared with baseline, showed an increase in nHF (t = 2.54, p = 0.02); a decrease in nLF (t = 2.62, p = 0.01) and in the LF/HF ratio (t = 4.00, p = 0.0003). In subgroup-C, at both 6 and 13-month follow-ups, the spectral components in the frequency domain HRV parameters were higher than baseline in nHF (t = 2.64, p = 0.02 and (t = 2.13, p = 0.05, respectively); lower in nLF (t = 2.64, p = 0.02 and (t = 2.13, p = 0.05, respectively), and in LF/HF (t = 1.92, p = 0.08 and (t = 2.43, p = 0.03, respectively). A significant proportion of HCWs reported persistent COVID-19 symptoms at both the 6 and 13-month follow-ups, seemingly unrelated to cardiac autonomic balance. In group 2 HCWs, at 1-month follow-up compared with baseline, the spectral components in the frequency domain HRV parameters, showed a decrease in nHF (t = 2.19, p = 0.04); an increase in nLF (t = 2.15, p = 0.04) and in LF/HF (t = 3.49, p = 0.002). Conclusion: These results are consistent with epidemiological data suggesting a higher risk of acute cardiovascular complications during the first 30 days after COVID-19. The SARS-CoV-2 associated autonomic imbalance in the post-acute phase after recovery of mild COVID-19 resolved 6 months after the first negative SARS-CoV-2 NPS. However, a significant proportion of HCWs reported long-term COVID-19 symptoms, which dot not seems to be related to cardiac autonomic balance. Future research should certainly further test whether autonomic imbalance has a role in the mechanisms of long-COVID syndrome.

Challenges and opportunities for SERS in the infrared: materials and methods.

In the wake of a global, heightened interest towards biomarker and disease detection prompted by the SARS-CoV-2 pandemic, surface enhanced Raman spectroscopy (SERS) positions itself again at the forefront of biosensing innovation. But is it ready to move from the laboratory to the clinic? This review presents the challenges associated with the application of SERS to the biomedical field, and thus, to the use of excitation sources in the near infrared, where biological windows allow for cell and through-tissue measurements. Two main tackling strategies will be discussed: (1) acting on the design of the enhancing substrate, which includes manipulation of nanoparticle shape, material, and supramolecular architecture, and (2) acting on the spectral collection set-up. A final perspective highlights the upcoming scientific and technological bets that need to be won in order for SERS to stably transition from benchtop to bedside.

The impact of SARS-CoV-2 on healthcare workers of a large University Hospital in the Veneto Region: risk of infection and clinical presentation in relation to different pandemic phases and some relevant determinants.

Aim: The aim of this study is to evaluate the incidence of SARS-CoV-2 infection and the prevalence of COVID-19-related symptoms in relation to pandemic phases and some relevant variables in a cohort of 8,029 HCWs from one of the largest Italian University Hospitals. Methods: A single-center retrospective study was performed on data collected during SARS-CoV-2 infection surveillance of HCWs. Cox's multiple regression was performed to estimate hazard ratios of SARS-CoV-2 infection. Logistic multivariate regression was used to assess the risk of asymptomatic infections and the onset of the most frequent symptoms. All analyses were adjusted for sociodemographic and occupational factors, pandemic phases, vaccination status, and previous infections. Results: A total of 3,760 HCWs resulted positive (2.0%-18.6% across five study phases). The total incidence rate of SARS-CoV-2 infection was 7.31 cases per 10,000 person-days, significantly lower in phase 1 and higher in phases 4 and 5, compared to phase 3. Younger HCWs, healthcare personnel, and unvaccinated subjects showed a higher risk of infection. Overall, 24.5% were asymptomatic infections, with a higher probability for men, physicians, and HCWs tested for screening, fully vaccinated, and those with previous infection. The clinical presentation changed over the phases in relation to vaccination status and the emergence of new variants. Conclusion: The screening activities of HCWs allowed for the early detection of asymptomatic cases, limiting the epidemic clusters inside the hospital wards. SARS-CoV-2 vaccination reduced infections and symptomatic cases, demonstrating again its paramount value as a preventive tool for occupational and public health.

Development of coronary dysfunction in adult progeny after maternal engineered nanomaterial inhalation during gestation.

Maternal exposure to environmental contaminants during pregnancy can profoundly influence the risk of developing cardiovascular disease in adult offspring. Our previous studies have demonstrated impaired cardiovascular health, microvascular reactivity, and cardiac function in fetal and young adult progeny after maternal inhalation of nano-sized titanium dioxide (nano-TiO2) aerosols during gestation. The present study was designed to evaluate the development of cardiovascular and metabolic diseases later in adulthood. Pregnant Sprague-Dawley rats were exposed to nano-TiO2 aerosols (~ 10 mg/m3, 134 nm median diameter) for 4 h per day, 5 days per week, beginning on gestational day (GD) 4 and ending on GD 19. Progeny were delivered in-house. Body weight was recorded weekly after birth. After 47 weeks, the body weight of exposed progeny was 9.4% greater compared with controls. Heart weight, mean arterial pressure, and plasma biomarkers of inflammation, dyslipidemia, and glycemic control were recorded at 3, 9 and 12 months of age, with no significant adaptations. While no clinical risk factors (i.e., hypertension, dyslipidemia, or systemic inflammation) emerged pertaining to the development of cardiovascular disease, we identified impaired endothelium-dependent and -independent arteriolar dysfunction and cardiac morphological alterations consistent with myocardial inflammation, degeneration, and necrosis in exposed progeny at 12 months. In conclusion, maternal inhalation of nano-TiO2 aerosols during gestation may promote the development of coronary disease in adult offspring.

Aptatag-based multiplexed assay for protein detection by surface-enhanced Raman spectroscopy.

Silver-nanoparticle dimers held together by a Raman reporter, capped with DNA aptamers and stabilized by polyethylene glycol chains, can be used to develop a multiplexed heterogeneous bioassay for protein detection with high sensitivity and selectivity.

Present and Future of Surface-Enhanced Raman Scattering.

The discovery of the enhancement of Raman scattering by molecules adsorbed on nanostructured metal surfaces is a landmark in the history of spectroscopic and analytical techniques. Significant experimental and theoretical effort has been directed toward understanding the surface-enhanced Raman scattering (SERS) effect and demonstrating its potential in various types of ultrasensitive sensing applications in a wide variety of fields. In the 45 years since its discovery, SERS has blossomed into a rich area of research and technology, but additional efforts are still needed before it can be routinely used analytically and in commercial products. In this Review, prominent authors from around the world joined together to summarize the state of the art in understanding and using SERS and to predict what can be expected in the near future in terms of research, applications, and technological development. This Review is dedicated to SERS pioneer and our coauthor, the late Prof. Richard Van Duyne, whom we lost during the preparation of this article.

Understanding the role of AgNO3 concentration and seed morphology in the achievement of tunable shape control in gold nanostars.

Gold nanostars are one of the most fascinating anisotropic nanoparticles. The morphology of a nanostar can be controlled by changing various synthetic parameters; however, the detailed growth mechanism is still not fully understood. Herein, we investigate this process in six-branched nanostars, focusing first on the properties of a single crystalline seed, which evolves to include penta-twinned defects as the gateway to anisotropic growth into the 6-branched morphology. In particular, we report on a high-yield seed-mediated protocol for the synthesis of these particles with high dimensional monodispersity in the presence of Triton-X, ascorbic acid, and AgNO3. Detailed spectroscopic and microscopic analyses have allowed the identification of several key intermediates in the growth process, revealing that it proceeds via penta-twinned intermediate seeds. Importantly, we report the first experimental evidence tracking the location of silver with sub-nanometer resolution and prove its role as a stabilizing agent in these highly branched nanostructures. Our results indicate that metallic silver on the spikes stabilizes the nanostar morphology and the remaining silver, present when AgNO3 is added at a high concentration, deposits on the core and between the bases of neighboring spikes. Importantly, we also demonstrate the possibility of achieving dimensional monodispersity, reproducibility, and tunability in colloidal gold nanostars that are substantially higher than those previously reported, which could be leveraged to carry out holistic computational-experimental studies to understand, predict, and tailor their plasmonic response.