Annual Review of Analytical Chemistry - Volume 16, 2023

Offering superb speed, chemical specificity, and analytical sensitivity, direct mass spectrometry (MS) technologies are highly amenable for the molecular analysis of complex tissues to aid in disease characterization and help identify new diagnostic, prognostic, and predictive markers. By enabling detection of clinically actionable molecular profiles from tissues and cells, direct MS technologies have the potential to guide treatment decisions and transform sample analysis within clinical workflows. In this review, we highlight recent health-related developments and applications of direct MS technologies that exhibit tangible potential to accelerate clinical research and disease diagnosis, including oncological and neurodegenerative diseases and microbial infections. We focus primarily on applications that employ direct MS technologies for tissue analysis, including MS imaging technologies to map spatial distributions of molecules in situ as well as handheld devices for rapid in vivo and ex vivo tissue analysis.

Recent developments in ion mobility (IM) technology have expanded the capability to separate and characterize gas-phase ions of biomolecules, especially when paired with mass spectrometry. This next generation of IM technology has been ushered in by creative innovation focused on both instrument architectures and how electric fields are applied. In this review, we focus on the application of high-resolution and multidimensional IM to biomolecular analyses, encompassing the fields of glycomics, lipidomics, peptidomics, and proteomics. We highlight selected research that demonstrates the application of the new IM toolkit to challenging biomolecular systems. Through our review of recently published literature, we outline the current strengths of respective technologies and perspectives for future applications.

Label-free electrochemical biosensing leverages the advantages of label-free techniques, low cost, and fewer user steps, with the sensitivity and portability of electrochemical analysis. In this review, we identify four label-free electrochemical biosensing mechanisms: (a) blocking the electrode surface, (b) allowing greater access to the electrode surface, (c) changing the intercalation or electrostatic affinity of a redox probe to a biorecognition unit, and (d) modulating ion or electron transport properties due to conformational and surface charge changes. Each mechanism is described, recent advancements are summarized, and relative advantages and disadvantages of the techniques are discussed. Furthermore, two avenues for gaining further diagnostic information from label-free electrochemical biosensors, through multiplex analysis and incorporating machine learning, are examined.

Electrochemical additive manufacturing is an advanced microfabrication technology capable of producing features of almost unlimited geometrical complexity. A unique combination of the capacity to process conductive materials, design freedom, and micro- to nanoscale resolution offered by these electrochemical techniques promises tremendous opportunities for a multitude of future applications spanning microelectronics, sensing, robotics, and energy storage. This review aims to equip readers with the basic principles of electrochemical 3D printing at the small length scale. By describing the basic principles of electrochemical additive manufacturing technology and using the recent advances in the field, this beginner's guide illustrates how controlling the fundamental phenomena that underpin the print process can be used to vary dimensions, morphology, and microstructure of printed structures.

Developing a deeper understanding of dynamic chemical, electronic, and morphological changes at interfaces is key to solving practical issues in electrochemical energy storage systems (EESSs). To unravel this complexity, an assortment of tools with distinct capabilities and spatiotemporal resolutions have been used to creatively visualize interfacial processes as they occur. This review highlights how electrochemical scanning probe techniques (ESPTs) such as electrochemical atomic force microscopy, scanning electrochemical microscopy, scanning ion conductance microscopy, and scanning electrochemical cell microscopy are uniquely positioned to address these challenges in EESSs. We describe the operating principles of ESPTs, focusing on the inspection of interfacial structure and chemical processes involved in Li-ion batteries and beyond. We discuss current examples, performance limitations, and complementary ESPTs. Finally, we discuss prospects for imaging improvements and deep learning for automation. We foresee that ESPTs will play an enabling role in advancing EESSs as we transition to renewable energies.

Neglected tropical diseases (NTDs) affect tropical and subtropical countries and are caused by viruses, bacteria, protozoa, and helminths. These kinds of diseases spread quickly due to the tropical climate and limited access to clean water, sanitation, and health care, which make exposed people more vulnerable. NTDs are reported to be difficult and inefficient to diagnose. As mentioned, most NTDs occur in countries that are socially vulnerable, and the lack of resources and access to modern laboratories and equipment intensify the difficulty of diagnosis and treatment, leading to an increase in the mortality rate. Portable and low-cost microfluidic systems have been widely applied for clinical diagnosis, offering a promising alternative that can meet the needs for fast, affordable, and reliable diagnostic tests in developing countries. This review provides a critical overview of microfluidic devices that have been reported in the literature for the detection of the most common NTDs over the past 5 years.

Biofilms are multicellular communities held together by a self-produced extracellular matrix and exhibit a set of properties that distinguish them from free-living bacteria. Biofilms are exposed to a variety of mechanical and chemical cues resulting from fluid motion and mass transport. Microfluidics provides the precise control of hydrodynamic and physicochemical microenvironments to study biofilms in general. In this review, we summarize the recent progress made in microfluidics-based biofilm research, including understanding the mechanism of bacterial adhesion and biofilm development, assessment of antifouling and antimicrobial properties, development of advanced in vitro infection models, and advancement in methods to characterize biofilms. Finally, we provide a perspective on the future direction of microfluidics-assisted biofilm research.

Gel matrices are fundamental to electrophoresis analyses of biopolymers in microscale channels. Both capillary gel and microchannel gel electrophoresis systems have produced fundamental advances in the scientific community. These analytical techniques remain as foundational tools in bioanalytical chemistry and are indispensable in the field of biotherapeutics. This review summarizes the current state of gels in microscale channels and provides a brief description of electrophoretic transport in gels. In addition to the discussion of traditional polymers, several nontraditional gels are introduced. Advances in gel matrices highlighted include selective polymers modified to contain added functionality as well as thermally responsive gels formed through self-assembly. This review discusses cutting-edge applications to challenging areas of discovery in DNA, RNA, protein, and glycan analyses. Finally, emerging techniques that result in multifunctional assays for real-time biochemical processing in capillary and three-dimensional channels are identified.

The development of wearable devices provides approaches for the realization of self-health care. Easily carried wearable devices allow individual health monitoring at any place whenever necessary. There are various interesting monitoring targets, including body motion, organ pressure, and biomarkers. An efficient use of space in one small device is a promising resolution to increase the functions of wearable devices. Through integration of a microfluidic system into wearable devices, embedding complicated structures in one design becomes possible and can enable multifunction analyses within a limited device volume. This article reviews the reported microfluidic wearable devices, introduces applications to different biofluids, discusses characteristics of the design strategies and sensing principles, and highlights the attractive configurations of each device. This review seeks to provide a detailed summary of recent advanced microfluidic wearable devices. The overview of advanced key components is the basis for the development of future microfluidic wearable devices.

Infrared (IR) spectroscopic imaging records spatially resolved molecular vibrational spectra, enabling a comprehensive measurement of the chemical makeup and heterogeneity of biological tissues. Combining this novel contrast mechanism in microscopy with the use of artificial intelligence can transform the practice of histopathology, which currently relies largely on human examination of morphologic patterns within stained tissue. First, this review summarizes IR imaging instrumentation especially suited to histopathology, analyses of its performance, and major trends. Second, an overview of data processing methods and application of machine learning is given, with an emphasis on the emerging use of deep learning. Third, a discussion on workflows in pathology is provided, with four categories proposed based on the complexity of methods and the analytical performance needed. Last, a set of guidelines, termed experimental and analytical specifications for spectroscopic imaging in histopathology, are proposed to help standardize the diversity of approaches in this emerging area.

In recent years, there has been a revived appreciation for the importance of spatial context and morphological phenotypes for both understanding disease progression and guiding treatment decisions. Compared with conventional 2D histopathology, which is the current gold standard of medical diagnostics, nondestructive 3D pathology offers researchers and clinicians the ability to visualize orders of magnitude more tissue within their natural volumetric context. This has been enabled by rapid advances in tissue-preparation methods, high-throughput 3D microscopy instrumentation, and computational tools for processing these massive feature-rich data sets. Here, we provide a brief overview of many of these technical advances along with remaining challenges to be overcome. We also speculate on the future of 3D pathology as applied in translational investigations, preclinical drug development, and clinical decision-support assays.

Since the early 1990s, single-molecule detection in solution at room temperature has enabled direct observation of single biomolecules at work in real time and under physiological conditions, providing insights into complex biological systems that the traditional ensemble methods cannot offer. In particular, recent advances in single-molecule tracking techniques allow researchers to follow individual biomolecules in their native environments for a timescale of seconds to minutes, revealing not only the distinct pathways these biomolecules take for downstream signaling but also their roles in supporting life. In this review, we discuss various single-molecule tracking and imaging techniques developed to date, with an emphasis on advanced three-dimensional (3D) tracking systems that not only achieve ultrahigh spatiotemporal resolution but also provide sufficient working depths suitable for tracking single molecules in 3D tissue models. We then summarize the observables that can be extracted from the trajectory data. Methods to perform single-molecule clustering analysis and future directions are also discussed.

The goal of protecting the health of future generations is a blueprint for future biosensor design. Systems-level decision support requires that biosensors provide meaningful service to society. In this review, we summarize recent developments in cyber physical systems and biosensors connected with decision support. We identify key processes and practices that may guide the establishment of connections between user needs and biosensor engineering using an informatics approach. We call for data science and decision science to be formally connected with sensor science for understanding system complexity and realizing the ambition of biosensors-as-a-service. This review calls for a focus on quality of service early in the design process as a means to improve the meaningful value of a given biosensor. We close by noting that technology development, including biosensors and decision support systems, is a cautionary tale. The economics of scale govern the success, or failure, of any biosensor system.

Biosensing based on CRISPR-Cas systems is a young but rapidly evolving technology. The unprecedented properties of the CRISPR-Cas system provide an innovative tool for developing new-generation biosensing strategies. To date, a series of nucleic acid and non-nucleic acid detection methods have been developed based on the CRISPR platform. In this review, we first introduce the core biochemical properties underpinning the development of CRISPR bioassays, such as diverse reaction temperatures, programmability in design, high reaction efficiency, and recognition specificity, and highlight recent efforts to improve these parameters. We then introduce the technical developments, including how to improve sensitivity and quantification capabilities, develop multiplex assays, achieve convenient one-pot assays, create advanced sensors, and extend the applications of detection. Finally, we analyze obstacles to the commercial application of CRISPR detection technology and explore development opportunities and directions.

Cell functions rely on signal transduction—the cascades of molecular interactions and biochemical reactions that relay extracellular signals to the cell interior. Dissecting principles governing the signal transduction process is critical for the fundamental understanding of cell physiology and the development of biomedical interventions. The complexity of cell signaling is, however, beyond what is accessible by conventional biochemistry assays. Thanks to their unique physical and chemical properties, nanoparticles (NPs) have been increasingly used for the quantitative measurement and manipulation of cell signaling. Even though research in this area is still in its infancy, it has the potential to yield new, paradigm-shifting knowledge of cell biology and lead to biomedical innovations. To highlight this importance, we summarize in this review studies that pioneered the development and application of NPs for cell signaling, from quantitative measurements of signaling molecules to spatiotemporal manipulation of cell signal transduction.

This review summarizes the current status of development in photoluminescent probes, multidimensional photoluminescence detection, and multivariate data analysis methods. It then highlights reports featuring multivariate analysis of multidimensional measurements of photoluminescent probes published between June 2015 and June 2022, emphasizing work in the last 5 years. Important trends include the development of probe arrays, which provide fingerprint responses to the analyte(s) of interest and facilitate the analysis of complex samples; the application of neural networks and deep learning to pattern recognition and feature selection in photoluminescence images; and the application of multiway multivariate analysis to mining matrices, three-way arrays, and higher-order measurements, including hyperspectral intensity and lifetime images. These examples illustrate the increase in information extraction provided by the combination of multidimensional measurements and multivariate analysis.

The identification of thousands of proteins and their relative levels of expression has furthered understanding of biological processes and disease and stimulated new systems biology hypotheses. Quantitative proteomics workflows that rely on analytical assays such as mass spectrometry have facilitated high-throughput measurements of proteins partially due to multiplexing. Multiplexing allows proteome differences across multiple samples to be measured simultaneously, resulting in more accurate quantitation, increased statistical robustness, reduced analysis times, and lower experimental costs. The number of samples that can be multiplexed has evolved from as few as two to more than 50, with studies involving more than 10 samples being denoted as enhanced multiplexing or hyperplexing. In this review, we give an update on emerging multiplexing proteomics techniques and highlight advantages and limitations for enhanced multiplexing strategies.

This article critically reviews analytical method validation and quality control applied to the environmental chemistry field. The review focuses on the determination of organic micropollutants (OMPs), specifically emerging contaminants and pesticides, in the aquatic environment. The analytical technique considered is (gas and liquid) chromatography coupled to mass spectrometry (MS), including high-resolution MS for wide-scope screening purposes. An analysis of current research practices outlined in the literature has been performed, and key issues and analytical challenges are identified and critically discussed. It is worth emphasizing the lack of specific guidelines applied to environmental analytical chemistry and the minimal regulation of OMPs in waters, which greatly affect method development and performance, requirements for method validation, and the subsequent application to samples. Finally, a proposal is made for method validation and data reporting, which can be understood as starting points for further discussion with specialists in environmental analytical chemistry.

Despite the fact that oil chemistry and oils spills have been studied for many years, there are still emerging techniques and unknown processes to be explored. The 2010 Deepwater Horizon oil spill in the Gulf of Mexico resulted in a revival of oil spill research across a wide range of fields. These studies provided many new insights, but unanswered questions remain. Over 1,000 journal articles related to the Deepwater Horizon spill are indexed by the Chemical Abstract Service. Numerous ecological, human health, and organismal studies were published. Analytical tools applied to the spill include mass spectrometry, chromatography, and optical spectroscopy. Owing to the large scale of studies, this review focuses on three emerging areas that have been explored but remain underutilized in oil spill characterization: excitation-emission matrix spectroscopy, black carbon analysis, and trace metal analysis using inductively coupled plasma mass spectrometry.

In situ water monitoring sensors are critical to gain an understanding of ocean biochemistry and ecosystem health. They enable the collection of high-frequency data and capture ecosystem spatial and temporal changes, which in turn facilitate long-term global predictions. They are used as decision support tools in emergency situations and for risk mitigation, pollution source tracking, and regulatory monitoring. Advanced sensing platforms exist to support various monitoring needs together with state-of-the-art power and communication capabilities. To be fit-for-purpose, sensors must withstand the challenging marine environment and provide data at an acceptable cost. Significant technological advancements have catalyzed the development of new and improved sensors for coastal and oceanographic applications. Sensors are becoming smaller, smarter, more cost-effective, and increasingly specialized and diversified. This article, therefore, provides a review of the state-of-the art oceanographic and coastal sensors. Progress in sensor development is discussed in terms of performance and the key strategies used for achieving robustness, marine rating, cost reduction, and antifouling protection.