Fluoride-assisted detection of glutathione by surface Ce3+/Ce4+ engineered nanoceria.

Nanoceria has evolved as a promising nanomaterial due to its unique enzyme-like properties, including excellent oxidase mimetic activity, which significantly increases in the presence of fluoride ions. However, this significant increase in oxidase activity has never been utilised as a signal enhancer for the detection of biological analytes partly because of the lack of understanding of the mechanism involved in this process. In this study, we show that the surface oxidation state of cerium ions plays a very crucial role in different enzymatic activities, especially the oxidase mimetic activity by engineering nanoceria with three different surface Ce4+/Ce3+ compositions. Using DFT calculations combined with Bader charge analysis, it is demonstrated that stoichiometric ceria registers a higher oxidase mimetic activity than oxygen-deficient ceria with a low Ce4+/Ce3+ ratio due to a higher charge transfer from a substrate, 3,3',5,5' tetramethylbenzidine (TMB), to the ceria surface. We also show that the fluoride ions can significantly increase the charge transfer from the TMB surface to ceria irrespective of the surface Ce4+/Ce3+ ratio. Using this knowledge, we first compare the fluoride sensing properties of nanoceria with high Ce4+ and mixed Ce4+/Ce3+ oxidation states and further demonstrate that the linear detection range of fluoride ions can be extended to 1-10 ppm for nanoceria with mixed oxidation states. Then, we also demonstrate an assay for fluoride assisted detection of glutathione, an antioxidant with elevated levels during cancer, using nanoceria with a high surface Ce4+/Ce3+ ratio. The addition of fluoride ions in this assay allows the detection of glutathione in the linear range of 2.5-50 ppm with a limit of detection (LOD) of 3.8 ppm. These studies not only underpin the role of the surface Ce4+/Ce3+ ratio in tuning the fluoride assisted boost in the oxidase mimetic activity of nanoceria but also its strategic application in designing better colourimetric assays.

Tuning the enzyme-like activities of cerium oxide nanoparticles using a triethyl phosphite ligand.

Cerium oxide nanoparticles (CeNPs) exhibit excellent in vitro and in vivo antioxidant properties, determined by the redox switching of surface cerium ions between their two oxidation states (Ce3+ and Ce4+). It is known that ligands such as triethyl phosphite (TEP) can tune the redox behavior of CeNPs and change their biological enzyme-mimetic activities; however, the corresponding mechanism for such a behavior is completely unknown. Herein, we have studied the effect of TEP in promoting the SOD-enzyme-like activity in CeNPs with high and low Ce3+/Ce4+ ratio, which were synthesized by wet chemical and thermal hydrolysis methods, respectively, and incubated with varying concentrations of TEP. X-ray diffraction, UV-visible, photoluminescence, X-ray photoelectron spectroscopy, and Raman spectroscopy combined with DFT calculations were used to investigate the interaction of TEP on the surface of CeNPs. We observed a clear correlation between TEP concentration and the formation of surface oxygen vacancies. XPS analysis confirmed the increase in Ce3+ concentration after interaction with TEP. Moreover, we show that TEP's influence depends on the surface Ce3+/Ce4+ ratio. The superoxide dismutase-, catalase-, and oxidase-like activities of CeNPs with high Ce3+/Ce4+ ratio are not affected by TEP interaction, whereas catalase- and oxidase-like activities of CeNPs with low Ce3+/Ce4+ ratio decrease and the SOD-like activity is found to increase upon incubation with different concentrations of TEP. We also demonstrate that TEP interaction does not affect the regeneration of the CeNP surface, while the DFT calculations show that TEP facilitates the formation of defects on the surface of stoichiometric cerium oxide by reducing the oxygen vacancy formation energy. CeNPs with low Ce3+/Ce4+ ratio incubated with TEP also exhibited good antibacterial activity as compared to the CeNPs or TEP alone.

Chemical bond imaging using torsional and flexural higher eigenmodes of qPlus sensors.

Non-contact atomic force microscopy (AFM) with CO-functionalized tips allows visualization of the chemical structure of adsorbed molecules and identify individual inter- and intramolecular bonds. This technique enables in-depth studies of on-surface reactions and self-assembly processes. Herein, we analyze the suitability of qPlus sensors, which are commonly used for such studies, for the application of modern multifrequency AFM techniques. Two different qPlus sensors were tested for submolecular resolution imaging via actuating torsional and flexural higher eigenmodes and via bimodal AFM. The torsional eigenmode of one of our sensors is perfectly suited for performing lateral force microscopy (LFM) with single bond resolution. The obtained LFM images agree well with images from the literature, which were scanned with customized qPlus sensors that were specifically designed for LFM. The advantage of using a torsional eigenmode is that the same molecule can be imaged either with a vertically or laterally oscillating tip without replacing the sensor simply by actuating a different eigenmode. Submolecular resolution is also achieved by actuating the 2nd flexural eigenmode of our second sensor. In this case, we observe particular contrast features that only appear in the AFM images of the 2nd flexural eigenmode but not for the fundamental eigenmode. With complementary laser Doppler vibrometry measurements and AFM simulations we can rationalize that these contrast features are caused by a diagonal (i.e. in-phase vertical and lateral) oscillation of the AFM tip.

Experimental analysis of tip vibrations at higher eigenmodes of QPlus sensors for atomic force microscopy.

QPlus sensors are non-contact atomic force microscope probes constructed from a quartz tuning fork and a tungsten wire with an electrochemically etched tip. These probes are self-sensing and offer an atomic-scale spatial resolution. Therefore, qPlus sensors are routinely used to visualize the chemical structure of adsorbed organic molecules via the so-called bond imaging technique. This is achieved by functionalizing the AFM tip with a single CO molecule and exciting the sensor at the first vertical cantilever resonance mode. Recent work using higher-order resonance modes has also resolved the chemical structure of single organic molecules. However, in these experiments, the image contrast can differ significantly from the conventional bond imaging contrast, which was suspected to be caused by unknown vibrations of the tip. This work investigates the source of these artefacts by using a combination of mechanical simulation and laser vibrometry to characterize a range of sensors with different tip wire geometries. The results show that increased tip mass and length cause increased torsional rotation of the tuning fork beam due to the off-center mounting of the tip wire, and increased flexural vibration of the tip. These undesirable motions cause lateral deflection of the probe tip as it approaches the sample, which is rationalized to be the cause of the different image contrast. The results also provide a guide for future probe development to reduce these issues.

A review of demodulation techniques for multifrequency atomic force microscopy.

This article compares the performance of traditional and recently proposed demodulators for multifrequency atomic force microscopy. The compared methods include the lock-in amplifier, coherent demodulator, Kalman filter, Lyapunov filter, and direct-design demodulator. Each method is implemented on a field-programmable gate array (FPGA) with a sampling rate of 1.5 MHz. The metrics for comparison include the sensitivity to other frequency components and the magnitude of demodulation artifacts for a range of demodulator bandwidths. Performance differences are demonstrated through higher harmonic atomic force microscopy imaging.

Multimodal atomic force microscopy with optimized higher eigenmode sensitivity using on-chip piezoelectric actuation and sensing.

Atomic force microscope (AFM) cantilevers with integrated actuation and sensing provide several distinct advantages over conventional cantilever instrumentation. These include clean frequency responses, the possibility of down-scaling and parallelization to cantilever arrays as well as the absence of optical interference. While cantilever microfabrication technology has continuously advanced over the years, the overall design has remained largely unchanged; a passive rectangular shaped cantilever design has been adopted as the industry wide standard. In this article, we demonstrate multimode AFM imaging on higher eigenmodes as well as bimodal AFM imaging with cantilevers using fully integrated piezoelectric actuation and sensing. The cantilever design maximizes the higher eigenmode deflection sensitivity by optimizing the transducer layout according to the strain mode shape. Without the need for feedthrough cancellation, the read-out method achieves close to zero actuator/sensor feedthrough and the sensitivity is sufficient to resolve the cantilever Brownian motion.

Lyapunov estimation for high-speed demodulation in multifrequency atomic force microscopy.

An important issue in the emerging field of multifrequency atomic force microscopy (MF-AFM) is the accurate and fast demodulation of the cantilever-tip deflection signal. As this signal consists of multiple frequency components and noise processes, a lock-in amplifier is typically employed for its narrowband response. However, this demodulator suffers inherent bandwidth limitations as high-frequency mixing products must be filtered out and several must be operated in parallel. Many MF-AFM methods require amplitude and phase demodulation at multiple frequencies of interest, enabling both z-axis feedback and phase contrast imaging to be achieved. This article proposes a model-based multifrequency Lyapunov filter implemented on a field-programmable gate array (FPGA) for high-speed MF-AFM demodulation. System descriptions and simulations are verified by experimental results demonstrating high tracking bandwidths, strong off-mode rejection and minor sensitivity to cross-coupling effects. Additionally, a five-frequency system operating at 3.5 MHz is implemented for higher harmonic amplitude and phase imaging up to 1 MHz.

A review of demodulation techniques for amplitude-modulation atomic force microscopy.

In this review paper, traditional and novel demodulation methods applicable to amplitude-modulation atomic force microscopy are implemented on a widely used digital processing system. As a crucial bandwidth-limiting component in the z-axis feedback loop of an atomic force microscope, the purpose of the demodulator is to obtain estimates of amplitude and phase of the cantilever deflection signal in the presence of sensor noise or additional distinct frequency components. Specifically for modern multifrequency techniques, where higher harmonic and/or higher eigenmode contributions are present in the oscillation signal, the fidelity of the estimates obtained from some demodulation techniques is not guaranteed. To enable a rigorous comparison, the performance metrics tracking bandwidth, implementation complexity and sensitivity to other frequency components are experimentally evaluated for each method. Finally, the significance of an adequate demodulator bandwidth is highlighted during high-speed tapping-mode atomic force microscopy experiments in constant-height mode.

Note: Guaranteed collocated multimode control of an atomic force microscope cantilever using on-chip piezoelectric actuation and sensing.

The quality (Q) factor is an important parameter of the resonance of the microcantilever as it determines both imaging bandwidth and force sensitivity. The ability to control the Q factor of multiple modes is believed to be of great benefit for atomic force microscopy techniques involving multiple eigenmodes. In this paper, we propose a novel cantilever design employing multiple piezoelectric transducers which are used for separated actuation and sensing, leading to guaranteed collocation of the first eight eigenmodes up to 3 MHz. The design minimizes the feedthrough usually observed with these systems by incorporating a guard trace on the cantilever chip. As a result, a multimode Q controller is demonstrated to be able to modify the quality factor of the first two eigenmodes over up to four orders of magnitude without sacrificing robust stability.

Multimodal cantilevers with novel piezoelectric layer topology for sensitivity enhancement.

Self-sensing techniques for atomic force microscope (AFM) cantilevers have several advantageous characteristics compared to the optical beam deflection method. The possibility of down scaling, parallelization of cantilever arrays and the absence of optical interference associated imaging artifacts have led to an increased research interest in these methods. However, for multifrequency AFM, the optimization of the transducer layout on the cantilever for higher order modes has not been addressed. To fully utilize an integrated piezoelectric transducer, this work alters the layout of the piezoelectric layer to maximize both the deflection of the cantilever and measured piezoelectric charge response for a given mode with respect to the spatial distribution of the strain. On a prototype cantilever design, significant increases in actuator and sensor sensitivities were achieved for the first four modes without any substantial increase in sensor noise. The transduction mechanism is specifically targeted at multifrequency AFM and has the potential to provide higher resolution imaging on higher order modes.

High-bandwidth multimode self-sensing in bimodal atomic force microscopy.

Using standard microelectromechanical system (MEMS) processes to coat a microcantilever with a piezoelectric layer results in a versatile transducer with inherent self-sensing capabilities. For applications in multifrequency atomic force microscopy (MF-AFM), we illustrate that a single piezoelectric layer can be simultaneously used for multimode excitation and detection of the cantilever deflection. This is achieved by a charge sensor with a bandwidth of 10 MHz and dual feedthrough cancellation to recover the resonant modes that are heavily buried in feedthrough originating from the piezoelectric capacitance. The setup enables the omission of the commonly used piezoelectric stack actuator and optical beam deflection sensor, alleviating limitations due to distorted frequency responses and instrumentation cost, respectively. The proposed method benefits from a more than two orders of magnitude increase in deflection to strain sensitivity on the fifth eigenmode leading to a remarkable signal-to-noise ratio. Experimental results using bimodal AFM imaging on a two component polymer sample validate that the self-sensing scheme can therefore be used to provide both the feedback signal, for topography imaging on the fundamental mode, and phase imaging on the higher eigenmode.

A novel self-sensing technique for tapping-mode atomic force microscopy.

This work proposes a novel self-sensing tapping-mode atomic force microscopy operation utilizing charge measurement. A microcantilever coated with a single piezoelectric layer is simultaneously used for actuation and deflection sensing. The cantilever can be batch fabricated with existing micro electro mechanical system processes. The setup enables the omission of the optical beam deflection technique which is commonly used to measure the cantilever oscillation amplitude. Due to the high amount of capacitive feedthrough in the measured charge signal, a feedforward control technique is employed to increase the dynamic range from less than 1 dB to approximately 35 dB. Experiments show that the conditioned charge signal achieves excellent signal-to-noise ratio and can therefore be used as a feedback signal for atomic force microscopy imaging.