Identification and quantification of pigments in plant leaves using thin layer chromatography-Raman spectroscopy (TLC-Raman).

Carotenoids are yellow, orange, and red pigments commonly found in plants. In leaves, these molecules are essential for photosynthesis, but they also play a major role in plant growth and development. Efficiently monitoring concentrations of specific carotenoids in plant tissues could help to explain plant responses to environmental stressors, infection and disease, fertilization, and other conditions. Previously, Raman methods have been used to demonstrate a correlation between plant fitness and the carotenoid content of leaves. Due to solvatochromatic effects and structural similarities within the carotenoid family, current Raman spectroscopy techniques struggle to assign signals to specific carotenoids with certainty, complicating the determination of amounts of individual carotenoids present in a sample. In this work, we use thin layer chromatography-Raman spectroscopy, or TLC-Raman, to identify and quantify carotenoids extracted from tomato leaves. These quick and accurate methods could be applied to study the relationship between pigment content and a number of factors affecting plant health.

From the lab to the field: handheld surface enhanced Raman spectroscopy (SERS) detection of viral proteins.

Translating sensors from the lab benchtop to a readily available point-of-need setting is desirable for many fields, including medicine, agriculture, and industry. However, this transition generally suffers from loss of sensitivity, high background signals, and other issues which can impair reproducibility. Here we adapt a label-free surface-enhanced Raman spectroscopy (SERS) sensor for SARS-CoV-2 antigens from a lab-based assay to a handheld device. Utilizing a peptide capture molecule, which we previously employed for a surface-based assay, we optimize a simpler and more cost-efficient nanoparticle-based assay. This new assay allows for the direct detection of these viral antigens by SERS, now with the advantages of robustness and portability. We highlight considerations for nanoparticle modification conditions and warn against methods which can interfere with accurate detection. The comparison of these two assays will help guide further development of SERS-based sensors into devices that can be easily used in point-of-care settings, such as by emergency room nurses, farmers, or quality control technicians.

Monitoring and Characterization of Milk Fouling on Stainless Steel Using a High-Pressure High-Temperature Quartz Crystal Microbalance with Dissipation.

Fouling at interfaces deteriorates the efficiency and hygiene of processes within numerous industrial sectors, including the oil and gas, biomedical device, and food industries. In the food industry, the fouling of a complex food matrix to a heated stainless steel surface reduces production efficiency by increasing heating resistance, pumping requirements, and the frequency of cleaning operations. In this work, quartz crystal microbalance with dissipation (QCM-D) was used to study the interface formed by the fouling of milk on a stainless steel surface at different flow rates and protein concentrations at high temperatures (135 °C). Subsequently, the QCM-D response was recorded during the cleaning of the foulant. Two phases of fouling were identified. During phase-1, the fouling rate was dependent on the flow rate, while the fouling rate during phase-2 was dependent on the flow rate and protein concentration. During cleaning, foulants deposited at the higher flow rate swelled more than those deposited at the lower flow rate. The composition of the fouling deposits consisted of both protein and mineral species. Two crystalline phases of calcium phosphate, ß-tricalcium phosphate and hydroxyapatite, were identified at both flow rates. Stratification in topography was observed across the surface of the QCM-D sensor with a brittle and cracked structure for deposits formed at 0.2 mL/min and a smooth and close-packed structure for deposits formed at 0.1 mL/min. These stratifications in the composition and topography were correlated to differences in the reaction time and flow dynamics at different flow rates. This high-temperature application of QCM-D to complex food systems illuminates the initial interaction between proteins and minerals and a stainless steel surface, which might otherwise be undetectable in low-temperature applications of QCM-D or at larger bench and industrial scales. The methods and results presented here have implications for optimizing processing scenarios that limit fouling formation while also enhancing removal during cleaning.

Catching COVID: Engineering Peptide-Modified Surface-Enhanced Raman Spectroscopy Sensors for SARS-CoV-2.

COVID-19 remains an ongoing issue across the globe, highlighting the need for a rapid, selective, and accurate sensor for SARS-CoV-2 and its emerging variants. The chemical specificity and signal amplification of surface-enhanced Raman spectroscopy (SERS) could be advantageous for developing a quantitative assay for SARS-CoV-2 with improved speed and accuracy over current testing methods. Here, we have tackled the challenges associated with SERS detection of viruses. As viruses are large, multicomponent species, they can yield different SERS signals, but also other abundant biomolecules present in the sample can generate undesired signals. To improve selectivity in complex biological environments, we have employed peptides as capture probes for viral proteins and developed an angiotensin-converting enzyme 2 (ACE2) mimetic peptide-based SERS sensor for SARS-CoV-2. The unique vibrational signature of the spike protein bound to the peptide-modified surface is identified and used to construct a multivariate calibration model for quantification. The sensor demonstrates a 300 nM limit of detection and high selectivity in the presence of excess bovine serum albumin. This work provides the basis for designing a SERS-based assay for the detection of SARS-CoV-2 as well as engineering SERS biosensors for other viruses in the future.

Raman spectroscopy and neuroscience: from fundamental understanding to disease diagnostics and imaging.

Neuroscience would directly benefit from more effective detection techniques, leading to earlier diagnosis of disease. The specificity of Raman spectroscopy is unparalleled, given that a molecular fingerprint is attained for each species. It also allows for label-free detection with relatively inexpensive instrumentation, minimal sample preparation, and rapid sample analysis. This review summarizes Raman spectroscopy-based techniques that have been used to advance the field of neuroscience in recent years.

Surface-enhanced spatially-offset Raman spectroscopy (SESORS) for detection of neurochemicals through the skull at physiologically relevant concentrations.

Detection techniques for neurotransmitters that are rapid, label-free, and non-invasive are needed to move towards earlier diagnosis of neurological disease. Surface-enhanced Raman spectroscopy (SERS) allows for sensitive and selective detection of target analytes. The combination of SERS with spatially offset Raman spectroscopy (SORS) in a technique termed surface enhanced spatially offset Raman spectroscopy (SESORS) permits a sensitive and selective detection of neurotransmitters through the skull. Here, we present the SESORS detection of individual neurotransmitters and mixtures of neurotransmitters at physiologically relevant concentrations, while also establishing limits of detection.

In Vitro and In Vivo SERS Biosensing for Disease Diagnosis.

For many disease states, positive outcomes are directly linked to early diagnosis, where therapeutic intervention would be most effective. Recently, trends in disease diagnosis have focused on the development of label-free sensing techniques that are sensitive to low analyte concentrations found in the physiological environment. Surface-enhanced Raman spectroscopy (SERS) is a powerful vibrational spectroscopy that allows for label-free, highly sensitive, and selective detection of analytes through the amplification of localized electric fields on the surface of a plasmonic material when excited with monochromatic light. This results in enhancement of the Raman scattering signal, which allows for the detection of low concentration analytes, giving rise to the use of SERS as a diagnostic tool for disease. Here, we present a review of recent developments in the field of in vivo and in vitro SERS biosensing for a range of disease states including neurological disease, diabetes, cardiovascular disease, cancer, and viral disease.