Sensitivity Enhancement Research Articles

Performance optimization of a SERF atomic magnetometer based on flat-top light beam

We explore the impact of pumping beams with different transverse intensity profiles on the performance of the spin-exchange relaxation-free (SERF) atomic magnetometers (AMs). We conduct experiments comparing the traditional Gaussian optically-pumped AM with that utilizing the flat-top optically-pumped (FTOP) method. Our findings reveal that the FTOP-based approach outperforms the conventional method, exhibiting a larger response, a narrower magnetic resonance linewidth, and a superior low-frequency noise performance. Specifically, the use of FTOP method leads to a 16% enhancement in average sensitivity within 1 Hz–30 Hz frequency range. Our research emphasizes the significance of achieving transverse polarization uniformity in AMs, providing insights for future optimization efforts and sensitivity improvements in miniaturized magnetometers.

A sensitive fluoroimmunoassay for quantitative detection of imidacloprid based on quantum dot-streptavidin conjugate

Imidacloprid (IMI), the most commonly used neonicotinoid, is widely present in both the environment and agro-products due to extensive and prolonged application, posing potential risks to ecological security and human health. This study introduced a sensitive and rapid fluorescence-linked immunosorbent assay, employing Quantum Dot-Streptavidin conjugate (QDs-SA-FLISA), for efficient monitoring of IMI residues in agro-products. Under optimized conditions, the QDs-SA-FLISA exhibited a half-maximal inhibition concentration (IC50) of 1.70 ng/mL and a limit of detection (LOD, IC20) of 0.5 ng/mL. Investigation into the sensitivity enhancement effect of the QDs-SA revealed that the sensitivity (IC50) of the QDs-SA-FLISA was 7.3 times higher than that of ELISA. The recoveries and relative standard deviation (RSD) ranged from 81.7 to 118.1 % and 0.5–9.4 %, respectively, for IMI in brown rice, tomato and pear. There was no significant difference in IMI residues obtained between QDs-SA-FLISA and UHPLC-MS/MS. Thus, the QDs-SA-FLISA represents a reliable approach for the quantitative determination of IMI in agro-products.

Laser-printed paper ELISA and hydroxyapatite immobilization for colorimetric congenital anomalies screening in saliva

BackgroundAlpha-fetoprotein (AFP) is a fetal protein that can indicate congenital anomalies such as Down syndrome and spinal canal blockage when detected at abnormal levels in pregnant women. Current AFP detection methods rely on invasive blood or serum samples, which require sophisticated equipment. From the many solutions proposed, colorimetric paper-based assays excel in point-of-care settings. The concept of paper-based ELISA (p-ELISA) enhances traditional methods, aligning with the ASSURED criteria for diagnostics in resource-limited regions. Despite success in microfluidic paper-based assay devices, laser printing remains underexplored for p-ELISA. Additionally, modifying the paper surface provides an additional layer of sensitivity enhancement. ResultsIn this study, we developed a novel laser-printed paper-based ELISA (LP-pELISA) for rapid, sensitive, and noninvasive detection of AFP in saliva samples. The LP-pELISA platform was fabricated by printing hydrophobic barriers on filter paper using a laser printer, followed by depositing hydroxyapatite (HAp) as an immobilization material for the antibodies. The colorimetric detection was achieved using AuNPs functionalized with anti-AFP antibodies and silver nitrate enhancement. The LP-pELISA exhibited a linear response for AFP detection in both buffer and saliva samples over a range of 1.0–800 ng mL−1, with a limit of detection (LOD) reaching 1.0 ng mL−1. The assay also demonstrated good selectivity, repeatability, reproducibility, and stability. The LP-pELISA was further validated by testing spiked human saliva samples, showing its potential for point-of-care diagnosis of congenital disabilities. SignificanceThe LP-pELISA is a noninvasive platform showcasing simplicity, cost-effectiveness, and user-friendliness, utilizing laser printing, hydroxyapatite modification, and saliva samples to efficiently detect AFP. Beyond its application for AFP, this method's versatility extends to other biomarkers, positioning it as a catalyst for the evolution of paper-based biosensors. The LP-pELISA holds promise as a transformative tool for point-of-care diagnostics, fostering advancements in healthcare with its innovative technology.

Determination of phenytoin at trace levels in domestic wastewater and synthetic urine samples by gas chromatography-mass spectrometry after its preconcentration by simple liquid-phase microextraction.

This work presents a sensitive and accurate analytical method for the determination of phenytoin at trace levels in domestic wastewater and synthetic urine samples by gas chromatography-mass spectrometry (GC-MS) after the metal sieve-linked double syringe liquid-phase microextraction (MSLDS-LPME) method. A metal sieve was produced in our laboratory in order to disperse water-immiscible extraction solvents into aqueous media. Univariate optimization studies for the selection of proper extraction solvent, extraction solvent volume, mixing cycle, and initial sample volume were carried out. Under the optimum MSLDS-LPME conditions, mass-based dynamic range, limit of quantitation (LOQ), limit of detection (LOD), and percent relative standard deviation (%RSD) for the lowest concentration in calibration plot were figured out to be 100.5-10964.2 μg kg-1, 150.6 μg kg-1, 45.2 μg kg-1, and 9.4%, respectively. Detection power was improved as 187.7-folds by the developed MSLDS-LPME-GC-MS system while enhancement in calibration sensitivity was recorded as 188.0-folds. In the final step of this study, the accuracy and applicability of the proposed system were tested by matrix matching calibration strategy. Percent recovery results for domestic wastewater and synthetic urine samples were calculated as 95.6-110.3% and 91.7-106.6%, respectively. These results proved the accuracy and applicability of the proposed preconcentration method, and the obtained analytical results showed the efficiency of the lab-made metal sieve apparatus.

Experimental assessment of a low-cost aberration-free compact double aperture four-hololens imaging system: application to the enhancement of sensitivity and accuracy of in-plane displacement measurement

In this work, an experimental analysis of hololens imaging configuration consisting of four holographic lenses has been carried out to realize Duffy’s double aperture speckle interferometer. It is demonstrated that using holographic lenses recorded for typical f-number in the four-hololens imaging system, the sensitivity of measurement is not limited due to the f-number of the lens. However, the sensitivity can be significantly enhanced by increasing the angle between the plane and spherical waves while recording the holographic lenses. In-plane displacements were measured using holographic lenses recorded at nearly equal f-number and with different beam angles between the plane and spherical waves. Experimental results show that holographic lenses recorded with a large beam angle between the plane and the spherical waves give a higher measurement sensitivity in comparison to those holographic lenses that were recorded with a lower beam angle at an equal f-number. This work shows that a four-hololens imaging system can be used advantageously as a low-cost alternative to Duffy’s double aperture interferometer to overcome the need for expensive diffraction-limited low f-number lenses for higher measurement accuracy and sensitivity.

Engineering biomimetic nanosystem targeting multiple tumor radioresistance hallmarks for enhanced radiotherapy.

Tumor cells establish a robust self-defense system characterized by hypoxia, antioxidant overexpression, DNA damage repair, and so forth to resist radiotherapy. Targeting one of these features is insufficient to overcome radioresistance due to the feedback mechanisms initiated by tumor cells under radiotherapy. Therefore, we herein developed an engineering biomimetic nanosystem (M@HHPt) masked with tumor cell membranes and loaded with a hybridized protein-based nanoparticle carrying oxygens (O2) and cisplatin prodrugs (Pt(IV)) to target multiple tumor radioresistance hallmarks for enhanced radiotherapy. After administration, M@HHPt actively targeted and smoothly accumulated in tumor cells by virtue of its innate homing abilities to realize efficient co-delivery of O2 and Pt(IV). O2 introduction induced hypoxia alleviation cooperated with Pt(IV) reduction caused glutathione consumption greatly amplified radiotherapy-ignited cellular oxidative stress. Moreover, the released cisplatin effectively hindered DNA damage repair by crosslinking with radiotherapy-produced DNA fragments. Consequently, M@HHPt-sensitized radiotherapy significantly suppressed the proliferation of lung cancer H1975 cells with an extremely high sensitizer enhancement ratio of 1.91 and the progression of H1975 tumor models with an excellent tumor inhibition rate of 94.7%. Overall, this work provided a feasible strategy for tumor radiosensitization by overcoming multiple radioresistance mechanisms.

17O Solid-State NMR Spectroscopy of Lipid Membranes.

Despite the limitations posed by poor sensitivity, studies have reported the unique advantages of 17O based NMR spectroscopy to study systems existing in liquid, solid, or semisolid states. 17O NMR studies have exploited the remarkable sensitivity of quadrupole coupling and chemical shift anisotropy tensors to the local environment in the characterization of a variety of intra- and intermolecular interactions and motion. Recent studies have considerably expanded the use of 17O NMR to study dynamic intermolecular interactions associated with some of the challenging biological systems under magic angle spinning (MAS) and aligned conditions. The very fast relaxing nature of 17O has been well utilized in cellular and in vivo MRS (magnetic resonance spectroscopy) and MRI (magnetic resonance imaging) applications. The main focus of this Review is to highlight the new developments in the biological solids with a detailed discussion for a few selected examples including membrane proteins and nanodiscs. In addition to the unique benefits and limitations, the remaining challenges to overcome, and the impacts of higher magnetic fields and sensitivity enhancement techniques are discussed.

Nano-Engineered HfO2-Au photonic sensor for ultra-sensitive refractive index detection

In this study, we introduce a novel hafnium dioxide (HfO2) and thin gold layer based photonic crystal fiber (HfAu-PCF) sensor for refractive index measurement and optimize in the spectrum of 1.3 µm to 1.6 µm. The proposed sensor incorporates a D-shaped geometry for enhanced light-matter interaction, and the core of the fiber is surrounded by periodic air holes to facilitate efficient coupling of the guided mode with surface plasmons. The proposed HfAu-PCF sensor is geometrically optimized using finite element method (FEM) by fine-tuning air hole diameter, core size, layer thickness dimensions of the fiber to achieve performance stability. Leveraging the unique optical properties of HfO2 and Au, the sensor demonstrates superior performance metrics, achieving confinement loss (<10−6 dB/m), propagation constant (β) ∼ 6 × 106, V-Parameter < 2.3, a peak relative sensitivity of 99.05 % across the tested wavelength of 1.3 µm to 1.6 µm. Our findings reveal that choosing suitable parameters could lead to further enhancements in sensitivity, making the sensor for applications in bio-sensing, environmental monitoring, and chemical analysis, etc.

Evaluación dinámica de las escalas CHA2DS2-VASc y HAS-BLED para predecir ictus isquémico y hemorragia mayor en pacientes con fibrilación auricular

Introduction and objectivesStroke and bleeding risks in atrial fibrillation (AF) are often assessed at baseline to predict outcomes years later. We investigated whether dynamic changes in CHA2DS2-VASc and HAS-BLED scores over time modify risk prediction. MethodsWe included patients with AF who were stable while taking vitamin K antagonists. During a 6-year follow-up, all ischemic strokes/transient ischemic attacks (TIAs) and major bleeding events were recorded. CHA2DS2-VASc and HAS-BLED were recalculated every 2-years and tested for clinical outcomes at 2-year periods. ResultsWe included 1361 patients (mean CHA2DS2-VASc and HAS-BLED 4.0±1.7 and 2.9±1.2). During the follow-up, 156 (11.5%) patients had an ischemic stroke/TIA and 269 (19.8%) had a major bleeding event. Compared with the baseline CHA2DS2-VASc, the CHA2DS2-VASc recalculated at 2 years had higher predictive ability for ischemic stroke/TIA during the period from 2 to 4 years. Integrated discrimination improvement (IDI) and net reclassification improvement (NRI) showed improvements in sensitivity and better reclassification. The CHA2DS2-VASc recalculated at 4 years had better predictive performance than the baseline CHA2DS2-VASc during the period from 4 to 6 years, with an improvement in IDI and an enhancement of the reclassification. The recalculated HAS-BLED at 2-years had higher predictive ability than the baseline score for major bleeding during the period from 2 to 4 years, with significant improvements in sensitivity and reclassification. A slight enhancement in sensitivity was observed with the HAS-BLED score recalculated at 4 years compared with the baseline score. ConclusionsIn AF patients, stroke and bleeding risks are dynamic and change over time. The CHA2DS2-VASc and HAS-BLED scores should be regularly reassessed, particularly for accurate stroke risk prediction.

Design of Pentacene Thin-Film Transistor Based Hydrogen Gas Sensor with High-K Dielectric Materials for High Sensitivity

Electrical properties of an organic field-effect transistor were modelled in top gate top contact (TGTC) geometry and H2 gas sensors were designed for increased sensitivity based on the structure. Safety concerns related to hydrogen usage must be addressed; these hazardous characteristics include a wide flammable range (4%–75%) that results in a rapid burning velocity, a low minimum ignition energy (0.017 mJ), a high heat of combustion (143 kJ g−1), and the high diffusivity of hydrogen gas (0.61 cm2 s−1 in the air). These characteristics make it impossible to control hydrogen combustion after a specific time. All simulations were performed in the Silvaco TCAD ATLAS tool. We analysed the driving principle of gas sensors and introduced gas sensing properties in OFET using platinum metal at the gate electrode for H2 gas detection. IOFF, ION, and VTH are sensitivity parameters that alter when the metalwork function of the gate changes with respect to the gas present on it. The designed sensor was analysed for different dielectric materials. Results demonstrate that the increase in sensitivity for OFET-based H2 sensors is 73.4%, 80.7%, 90.5%, and 95.6% when the work function changes by 50, 100, 150, and 200 meV for Pt gate electrodes with an increase in dielectric value of insulating layer from SiO2 (3.9) to La2O3 (27). Results were compared with the In1-xGaxAs CGNWFET-based H2 sensor as the work function varies at 200 meV,the sensitivity enhancement with OFET-based H2 sensors is 8.09%.

Analysis of the molecular alterations in cancer cells following nanotechnology-assisted targeted radiotherapy using Raman spectroscopy

The study unveiled an innovative strategy for precise radiation targeting in cancer treatment, along with the monitoring of molecular changes induced by this therapeutic approach. In this research, we explored the impact of administering anti-HER2-AgNPs nanoconjugates either individually or in conjunction with gamma irradiation on the viability of SKBR3 breast cancer cells. The utilization of nanoconjugates resulted in an enhancement of cellular sensitivity toward radiation. The viability of the cells exhibited a decline as the dose of irradiation increased, and this decrease was further exacerbated by the passage of time following irradiation. The analysis of RS revealed distinct cellular responses in varying conditions. The observed increase in SERS intensity, resulting from the increment in dose from 0 to 2 Gy, can be attributed to the probable upregulation of HER2 expression induced by irradiation. The observed decrease in SERS intensity at doses of 4 and 6 Gy can be attributed to the likely reduction in HER2 expression. It was illustrated that the analysis of Raman spectroscopy data can aid in the identification of radiation-induced biochemical alterations in cancer cells during the application of nanoconjugates-based radiotherapy. The findings revealed that nanoconjugates have the potential to enhance cellular sensitivity to radiation along with facilitating the detection of radiation-induced biochemical alterations within cancer cells.

Xylazine suppresses fentanyl consumption during self-administration and induces a unique sex-specific withdrawal syndrome that is not altered by naloxone in rats.

Prescription and illicit opioid use are a public health crisis, with the landscape shifting to fentanyl use. Since fentanyl is 100-fold more potent than morphine, its use is associated with a higher risk of fatal overdose that can be remediated through naloxone (Narcan) administration. However, recent reports indicate that xylazine, an anesthetic, is increasingly detected in accidental fentanyl overdose deaths. Anecdotal reports suggest that xylazine may prolong the fentanyl "high," alter the onset of fentanyl withdrawal, and increase resistance to naloxone-induced reversal of overdose. To date, no preclinical studies have evaluated the impacts of xylazine on fentanyl self-administration (SA; 2.5 μg/kg/infusion) or withdrawal to our knowledge. We established a rat model of xylazine/fentanyl co-SA and withdrawal and evaluated outcomes as a function of biological sex. When administered alone, chronic xylazine (2.5 mg/kg, intraperitoneal) induced unique sex-specific withdrawal symptomatology, whereby females showed delayed onset of signs and a possible enhancement of sensitivity to the motor-suppressing effects of xylazine. Xylazine reduced fentanyl consumption in both male and female rats regardless of whether it was experimenter-administered or added to the intravenous fentanyl product (0.05, 0.10, and 0.5 mg/kg/infusion) when compared to fentanyl SA alone. Interestingly, this effect was dose-dependent when self-administered intravenously. Naloxone (0.1 mg/kg, subcutaneous injection) did not increase somatic signs of fentanyl withdrawal, regardless of the inclusion of xylazine in the fentanyl infusion in either sex; however, somatic signs of withdrawal were higher across time points in females after xylazine/fentanyl co-SA regardless of naloxone exposure as compared to females following fentanyl SA alone. Together, these results indicate that xylazine/fentanyl co-SA dose-dependently suppressed fentanyl intake in both sexes and induced a unique withdrawal syndrome in females that was not altered by acute naloxone treatment. (PsycInfo Database Record (c) 2024 APA, all rights reserved).

Resonant Structures for Sensitivity Enhancement of Rydberg-Atom Microwave Receivers

Resonant Structures for Sensitivity Enhancement of Rydberg-Atom Microwave Receivers

Polydimethylsiloxane-coated Fiber Bragg Grating as a Bend Sensor

The fiber Bragg grating sensor is often demonstrated as a bend sensor. However, the demonstrated sensor in previous studies was limited to low sensitivity, thus reducing the sensing performance. Hence, this work demonstrated the polydimethylsiloxane-coated fiber Bragg grating sensor for sensitivity enhancement. The proposed sensor with a diameter of 5 cm was bent by adjusting the translation stage from 0.1 cm to 0.9 cm distance in forward and backward direction. The results showed that the Bragg wavelength shifted towards the longer wavelength region when the distance decreased. The polydimethylsiloxane-coated fiber Bragg grating sensor shows a sensitivity of 1743000 A.U, 8 times higher than the uncoated fiber Bragg sensor with a sensitivity of 284000 A.U. The presence of the polydimethylsiloxane layer has successfully improved the sensor performance.

Miniaturized flexible micro-tube plasma ionization source for the effective ionization of non-easily ionizable pesticides in food with liquid chromatography/mass spectrometry

In this article, we have studied the potential of flexible microtube plasma (FμTP) as ionization source for the liquid chromatography high-resolution mass spectrometry detection of non-easily ionizable pesticides (viz. nonpolar and non-ionizable by acid/basic moieties). Phthalimide-related compounds such as dicofol, dinocap, o-phenylphenol, captan, captafol, folpet and their metabolites were studied. Dielectric barrier discharge ionization (DBDI) was examined using two electrode configurations, including the miniaturized one based on a single high-voltage (HV) electrode and a virtual ground electrode configuration (FμTP), and also the two-ring electrode DBDI configuration. Different ionization pathways were observed to ionize these challenging, non-easily ionizable nonpolar compounds, involving nucleophilic substitutions and proton abstraction, with subtle differences in the spectra obtained compared with APCI. An average sensitivity increase of 5-fold was attained compared with the standard APCI source. In addition, more tolerance with matrix effects was observed in both DBDI sources. The importance of the data reported is not just limited to the sensitivity enhancement compared to APCI, but, more notably, to the ability to effectively ionize nonpolar, late-eluting (in reverse-phase chromatography) non-ionizable compounds. Besides o-phenylphenol ([M − H]-), all the parent species were efficiently ionized through different mechanisms involving bond cleavages through the effect of plasma reagent species or its combination with thermal degradation and subsequent ionization. This tool can be used to figure out overlooked nonpolar compounds in different environmental samples of societal interest through non-target screening (NTS) strategies.

Enhancing the Consistency and Performance of Graphene-Based Devices via Al Intermediate-Layer-Assisted Transfer and Patterning.

Graphene has garnered widespread attention, and its use is being explored for various electronic devices due to its exceptional material properties. However, the use of polymers (PMMA, photoresists, etc.) during graphene transfer and patterning processes inevitably leaves residues on graphene surface, which can decrease the performance and yield of graphene-based devices. This paper proposes a new transfer and patterning process that utilizes an Al intermediate layer to separate graphene from polymers. Through DFT calculations, the binding energy of graphene-Al was found to be only -0.48 eV, much lower than that of PMMA and photoresist with graphene, making it easier to remove Al from graphene. Subsequently, this was confirmed through XPS analysis. A morphological characterization demonstrated that the graphene patterns prepared using the Al intermediate layer process exhibited higher surface quality, with significantly reduced roughness. It is noteworthy that the devices obtained with the proposed method exhibited a notable enhancement in both consistency and sensitivity during electrical testing (increase of 67.14% in temperature sensitivity). The low-cost and pollution-free graphene-processing method proposed in this study will facilitate the further commercialization of graphene-based devices.

Selected Ion Monitoring for Orbitrap-Based Metabolomics.

Orbitrap mass spectrometry in full scan mode enables the simultaneous detection of hundreds of metabolites and their isotope-labeled forms. Yet, sensitivity remains limiting for many metabolites, including low-concentration species, poor ionizers, and low-fractional-abundance isotope-labeled forms in isotope-tracing studies. Here, we explore selected ion monitoring (SIM) as a means of sensitivity enhancement. The analytes of interest are enriched in the orbitrap analyzer by using the quadrupole as a mass filter to select particular ions. In tissue extracts, SIM significantly enhances the detection of ions of low intensity, as indicated by improved signal-to-noise (S/N) ratios and measurement precision. In addition, SIM improves the accuracy of isotope-ratio measurements. SIM, however, must be deployed with care, as excessive accumulation in the orbitrap of similar m/z ions can lead, via space-charge effects, to decreased performance (signal loss, mass shift, and ion coalescence). Ion accumulation can be controlled by adjusting settings including injection time and target ion quantity. Overall, we suggest using a full scan to ensure broad metabolic coverage, in tandem with SIM, for the accurate quantitation of targeted low-intensity ions, and provide methods deploying this approach to enhance metabolome coverage.

Design and Optimization of Silver Nanostructured Arrays in Plasmonic Metamaterials for Sensitive Imaging Applications

This paper delves into the strategic design and optimization of silver (Ag) nanostructured arrays within plasmonic metamaterials, targeting the enhancement of imaging sensitivity. Leveraging Finite-Difference Time-Domain (FDTD) simulations, our research rigorously compares various Ag nanostructured geometries, including nanospheres, nanocones, nanodisks, and nanocubes. The aim is to pinpoint configurations that significantly enhance electric field localization on the surfaces of nanostructures, a pivotal factor. The nanocube array exhibits superior field enhancement, particularly in narrow nanogaps, suggesting its suitability for high-sensitivity applications. Further exploration into nanocube arrays reveals the crucial role of nanogap size and spacer layer thickness in tuning the optical properties through the manipulation of Fabry–Pérot and mirror image modes in metal–insulator–metal (MIM) structures. By presenting a thorough analysis of these nanostructured arrays, the study not only contributes to our understanding of the fundamental principles governing plasmonic metamaterials but also provides a solid foundation for future innovation in highly sensitive imaging applications. It underscores the importance of nanostructure design and optimization in achieving significant improvements in the performance of plasmonic devices, marking a pivotal step forward in the field of nanophotonics and its application to sensitive imaging technologies.

Application of nanomaterials in tumor radio-sensitization

Cancer is a main reason of death worldwide, there was about 10million deaths caused by cancer in 2020. Radiotherapy, as a traditional method of treating cancer, uses radiation to kill the lesions of tumor patients. People develop radiation sensitizers to improve the treatment efficiency of radiotherapy and achieve better therapeutic effects through multiple collaborative treatments. Because nanomaterials have special physical and chemical properties, they may bring us novel methods to increase the effectiveness of treatment and reduces damage to the patient's body. People construct nanoplatforms loaded with multiple drugs to achieve a combination of radiation sensitization, toxicity reduction, and biocompatibility enhancement. At present, most nano radiation sensitizers are still in the development stage and have not been put into clinical use. The current research provides potential application prospects, but the existence of toxicity or other side effects cannot be ruled out, and the cost is also a problem that these drugs face before clinical use. This article mainly summarizes some methods of nanomaterial enhanced radiation therapy, research achievements in the past five years, and prospects for future development in this field.

DNP-assisted solid-state NMR enables detection of proteins at nanomolar concentrations in fully protonated cellular milieu.

With the sensitivity enhancements conferred by dynamic nuclear polarization (DNP), magic angle spinning (MAS) solid state NMR spectroscopy experiments can attain the necessary sensitivity to detect very low concentrations of proteins. This potentially enables structural investigations of proteins at their endogenous levels in their biological contexts where their native stoichiometries with potential interactors is maintained. Yet, even with DNP, experiments are still sensitivity limited. Moreover, when an isotopically-enriched target protein is present at physiological levels, which typically range from low micromolar to nanomolar concentrations, the isotope content from the natural abundance isotopes in the cellular milieu can outnumber the isotope content of the target protein. Using isotopically enriched yeast prion protein, Sup35NM, diluted into natural abundance yeast lysates, we optimized sample composition. We found that modest cryoprotectant concentrations and fully protonated environments support efficient DNP. We experimentally validated theoretical calculations of the limit of specificity for an isotopically enriched protein in natural abundance cellular milieu. We establish that, using pulse sequences that are selective for adjacent NMR-active nuclei, proteins can be specifically detected in cellular milieu at concentrations in the hundreds of nanomolar. Finally, we find that maintaining native stoichiometries of the protein of interest to the components of the cellular environment may be important for proteins that make specific interactions with cellular constituents.