Current and Future Applications of Bioanalytical Imaging

Introduction

Analytical instruments serve as invaluable tools in a wide range of scientific disciplines, from clinical research and drug development through to environmental monitoring and food safety.

Devoted to the advancement of healthcare and the world we live in, scientists across these fields rely on and routinely use analytical instruments to obtain quantitative and/or qualitative data that will enable accurate and effective analysis of the substances they are studying and testing.

The Japan Analytical Instruments Manufacturers Association (JAIMA), a company dedicated to promoting the advancement of analytical instrumentation defines these instruments as “appliances, tools, or devices that qualitatively and/or quantitatively measure the composition, properties, structure, status, etc., of substances.” The instruments are classified based on their application such as in laboratory experiments and research, monitoring of the environment, medical examinations and automation processes.

At this year’s Pittcon, in Orlando, FL, Feb 26 – March 1, attendees can listen to experts discuss the wide range of bioanalytical techniques that are currently in use or under development within the diverse scientific community.

Sessions will be held by leading researchers on high throughput and high-resolution techniques used in the fields of nanotechnology, molecular biology, biopharma, clinical medicine, molecular biology and many more.

This includes this years presenter for the Plenary Lecture, Dr Stefan Hell, who will be discussing the science and bioanalytical technology behind breaking the diffraction barrier in fluorescence microscopy.
In the following chapters some of these technologies and their applications will be discussed in more detail, as well as some of the highlights Pittcon attendees can expect to enjoy.

Just some examples of the bioanalytical imaging techniques that will be discussed at Pittcon include ambient ionization methods for the simplification of mass spectrometry (MS) analysis, surface plasmon resonance imaging (SPRI) for real-time molecule detection and electroparamagnetic resonance (EPR) spectroscopy for the detection of free radical and transition metals in pharmaceuticals.

A multitude of companies involved in the manufacture and supply of state-of -the-art, world-leading analytical instrumentation and technologies will be exhibiting at the event and demonstrating their latest products. With major industry players including Thermo Fisher Scientific, Shimadzu, Waters, Bruker, BioRad, Ametek, Zeiss, Metrohm, Wasatch, Hamamatsu, Hitachi and Zeiss, all attending this year’s symposium, Pittcon 2018 will provide an outstanding opportunity to hear about the latest advances in analytical instrumentation.

References


Chapter 1 – Imaging-based methods and strategies

Image-based strategies that can probe biological systems at the nanomolecular, molecular, cellular, tissue and small-animal level are rapidly advancing understanding within a wide range of scientific disciplines. Bioanalytical platforms that use multiplexing, single nanoparticle sensing, nanofluidics, and single-molecule detection are becoming invaluable tools in the study of biomolecules, single living cells, proteomics and genomics.

Also called laser-assisted microdissection (LMD), laser capture microdissection (LCM) is an isolation technique that separates specifics cells from a heterogenous population. Recently, several techniques have combined laser ablation (LA) with liquid capture for MS applications. A liquid-vortex capture (LVC) probe collects laser ablated material that dissolves and then undergoes electrospray ionization (ESI).

Pittcon attendees can listen to John Cahill (Oak Ridge National Laboratory) presenting an approach to substrate analysis that couples LMD with ESI-MS using an LVC probe. Cahill will discuss one sampling mode that captured 100% of the material, compared with only around 50% captured by LA. One microdissection instrument that can be used for such experiments is the Arcturus XT™ LCM System from Thermo Fisher Scientific, an exhibitor at this year’s Pittcon.

An important advancement in MS is the development of “ambient ionization methods,” which enable the ionization of samples lying outside of the vacuum system, thereby simplifying MS analysis and allowing rapid data acquisition. One example is desorption electrospray ionization (DESI), where electrically charged droplets are directly applied to a sample surface to generate ions of the sample’s constituents. Another example is paper spray ionization, where application of a high voltage to paper wetted with a solution generates analyte ions.

At Pittcon, Graham Cooks (Purdue University) will trace the evolution of ambient ionization methods, with emphasis placed on DESI and paper spray ionization, especially for quantitative analyses. Orbitrap-based mass spectrometers are used to perform these analyses,
an example being Thermo Fisher Scientific’s LTQ Orbitrap XL™ Hybrid Ion Trap-Orbitrap Mass Spectrometer.

A key technology in chemical imaging and spectroscopy is capillary electrophoresis (CE). Unlike traditional electrophoresis, which is not precise enough for small molecule analysis, CE uses the difference in charge-to-size ratio as the separating force and can be used for both small and large molecules. However, concerns have arisen over the quality of CE data. For example, poor sensitivity often results in the need for time-consuming and potentially result-distorting fluorescent labelling.
HORIBA Scientific, a leading manufacturer of high-precision instruments and exhibitor at Pittcon 2018, has overcome this problem by integrating powerful particle tracking and data analysis algorithms within CE systems. HORIBA’s HPCE-512 – High Performance Capillary Electrophoresis provides superb resolution, accuracy and detection levels.

At the conference, Edgar Arriaga (University of Minnesota) will describe how particle tracking and capillary cytometry can be used together to characterize lipid droplets, as well as his plans to expand the methodologies to other organelle types.

For single cell analysis by CE, accurate isolation of individual cells is essential, which can be achieved using instruments such as the S3ETM Cell Sorter from Bio-Rad laboratories, also exhibiting at Pittcon. This compact, automated bench-top device has a confirmed accuracy of 100% for single cell sorting.

Today, researchers prefer real-time, label-free, high-throughput technologies for the interrogation of chemical and biomolecular interactions. Ryan Bailey (University of Michigan) will describe silicon photonic sensor arrays for the label-free detection of interactions involving the regulation of blood coagulation and the recent progress made using microring resonators and supporting microfluidic tools. In 2010, Bailey reported on the use of silicon photonic microring resonators for the real-time detection of cellular cytokine secretion, an important advance in protein biomarker detection. Amongst the exhibitors at Pittcon, will be Microfluidics from IDEX Health & Science, host to an impressive portfolio of high-pressure homogenizers for sample preparation in silicon photonic sensor arrays.

Robert Corn (University of California) will talk about surface plasmon resonance imaging (SPRI), a label-free multiplexed detection tool that provides information on kinetic processes, binding affinity, analyte concentration and real-time molecule detection. In 2007, Corn published a paper on the detection of protein biomarkers at picomolar concentrations using SPRI measurements of RNA aptamer microarrays. Although highly sensitive, SPRI generally requires the fabrication of high fidelity microarrays of purified proteins or antibodies. Now, Corn will discuss on-chip fabrication strategies that enables microarray self-assembly on the SPRI chip for immediate use in SPRI surface bioaffinity measurements. HORIBA’s instrument range also includes SPRI instruments for multiplex label-free interaction analysis, providing ideal solutions for the simultaneous analysis of hundreds of molecular interactions.

The need for informative molecular fingerprints across a diverse range of molecules has kept the development of ion activation methods at the forefront of the MS field. Photodissociation MS is a technique that combines the ability of photons to activate and fragment ions with sensitive detection of the product ions by MS. Jennifer Brodbelt (University of Texas) will discuss the use of ultraviolet photodissociation (UVPD) for the fragmentation of biomolecules and how this MS/MS method benefits peptide and protein analysis. Also featuring in Thermo Fisher Scientific’s instrument portfolio is the Orbitrap Fusion Lumos Tribrid Mass Spectrometer, which now features an optional UVPD hardware module.

The main point of protein profiling is to find biomarkers that have expression levels associated with disease. This involves two approaches, namely top-down proteomics and bottom-up proteomics. Unlike bottom-up proteomics, which requires proteins to be fragmented into peptides prior to MS analysis, top-down proteomics analyses intact proteins, thereby affording a higher throughput.

Sophie Harvey (Ohio State University) will describe the application of ion mobility-MS coupled with surface induced dissociation (SID) for the analysis of membrane protein complex interactions and protein−lipid interactions. In a 2016 study, Harvey looked at noncovalent protein complexes subjected to extended Gas-Phase trapping followed by SID, to explore how the structure of proteins and their complexes are affected by the absence of a solvent and how this changes over time in the gas phase. The instrument used was a modified quadrupole ion mobility time-of-flight (Q-IM-TOF) instrument called the Synapt G2S, with a customized SID device installed before the IM chamber (SIDIM). The manufacturer − Waters Corp. – will be exhibiting at Pittcon.

Marco Arruda (Unicamp) will discuss the application of metallomics-based systems to biological systems. In a 2007 article, Arruda compared herbicide-tolerant transgenic soybeans with nontransgenic soybeans to see whether genetic modification affected the crop’s metallome.

Using a matrix-assisted laser desorption-ionization quadrupole-time of flight (MALDI-QTOF-MS) Premier mass spectrometer from Waters Corp., one protein was shown to display the most characteristic change in metal ion content, with higher Ca(II), Cu(II) and Fe(II) concentrations in transgenic soybeans.

Metals also play an important role in human biological processes. For example, MALDI-TOF MS/MS has been used to compare blood serum of patient treatments for a number of medical conditions. Examples of equipment from companies exhibiting at Pittcon that can be used for this type of experiment include BioRad’s flexible gel flexible drying systems, Bruker’s Ultraflex III-MALDI-TOF/TOF Mass Spectrometer and Bruker’s AnchorChip™ targets.

References

Pittcon Tracks

Bioanalytical & Life Science
Biological molecules and xenobiotics (drugs, toxins) and their metabolites; study of biological systems; biosensors; forensic science and toxicology
Cannabis & Psychedelic
Identification, quantitative measurement, extraction, and quality assurance of cannabis-based and psychedelic products
Environment & Energy
Environmental detection and monitoring; energy production and storage; sustainability, climate, and green chemistry; food science/safety and agriculture
Instrumentation & Nanoscience
Instrumentation, detection, and sensors; laboratory information systems, data analysis, and artificial intelligence; characterization and processing of nanomaterials; art and archeology
Pharmaceutical & Biologic
Evaluating chemical composition and properties/activities of medicinal drugs and biologics; high-throughput screening and process control; drug discovery and design; personal care and consumer products
Professional Development
Leadership and power/soft skills; career navigation, DEI (diversity, equity and inclusion), communication, and entrepreneurship; education and teaching and more

Chapter 2 – Molecules and Materials in Bioimaging and Biomonitoring

The ever-improving capability of bioanalytical techniques to extract novel and quantitative information, along with advances in fields such as nanotechnology and molecular biology is revolutionizing the analysis of living systems, the detection of diagnostic biomarkers and the evaluation of therapeutic effects in model systems. The scope of bioanalytical techniques also extends into fields such as forensics, environmental monitoring and food quality control.

In the pharmaceutical industry, characterizing a drug’s shelf life is crucial to ensuring its stability and therefore correct dosage and patient safety. The presence of free radicals and transition metals (paramagnetic species) is often what causes degradation of a drug’s active pharmaceutical ingredient (API). Electroparamagnetic resonance (EPR) spectroscopy is the only technique that unambiguously and non-invasively detects these species so that they can be quantified and monitored.

Exhibiting at this year’s Pittcon is Bruker, developer of the EMXnano benchtop EPR spectrometer. This instrument measures API degradation, determines its cause and predicts its long-term stability characteristics, providing an ideal solution for optimizing product stability.

In the clinical field, one challenge in the area of oncology biomarker research is translating the detection of new oncogenic or tumorigenic biomarkers into treatment approaches. Pittcon exhibitor Thermo Fisher Scientific provides versatile mass spectrometry (MS) solutions that help to detect and validate markers for early cancer detection, aid proteomics studies and monitor treatment response.

In the food industry, increasing demands relating to food safety have led to ever-more rigorous regulations. To address these demands, Pittcon exhibitors Waters, provide sensitive, reproducible and versatile solutions for the screening of pesticides, natural toxins, packaging contaminants and other adulterants.

Bioanalytical instrumentation is also widely used in forensics and toxicology studies for the determination of blood alcohol, abusive inhalants and other volatiles, and Pittcon exhibitor Teledyne Tekmar offers a range of solutions that forensic laboratories and coroner offices can use to ensure their data is precise, accurate and dependable.

Robert Clifford from Shimadzu will discuss bioanalytical techniques used in the cannabis testing industry for potency profiling and screening of pesticides, residual solvents, heavy metals, mycotoxins and moisture content. Cannabis testing was also discussed at last year’s Pittcon, where analytical methods were covered, along with the development of laboratory standards, cannabis extraction methods and cannabis detection to meet law enforcement needs.

A critical part of cannabis testing is determining cannabinoid potency, which involves the quantification of at least three major cannabinoids: THC, CBD and CBN. Shimadzu’s integrated high-performance liquid chromatography (HPLC) systems are ideal solutions for this, including the new i-Series instruments, which feature intuitive graphical user interfaces that even inexperienced staff can operate.

For pesticide testing, gas chromatograph-mass spectrometry (GC-MS) is the preferred platform. Shimadzu’s GCMS-QP2010 Ultra instrument can be used for this, as can the company’s GCMS-QP2010 SE. Herbicides can also be tested for on either of these instruments. For residual solvent testing, a headspace GC device can be used such as Shimadzu’s GC-2010 Plus with HS-20 Headspace Sampler.

Moisture content can be measured using Shimadzu’s MOC63u (and MOC120H), microorganisms can be detected using the company’s LC and LCMS systems and heavy metal testing can be performed using the AA-7000 with GFA-7000 or ICPE-9800.

The detection capabilities of bioanalytical techniques will also be discussed in the context of pathogenic outbreaks and the growing interest from farmers, governments and industry in rapid and quantitative in-field testing for the detection of bacteria in food and beverages.

In a 2010 paper, Frederique Deiss (Indiana University) and colleagues report on a simple, electroanalytical system based on the combination of a hand-held glucometer with micro-paper-based analytical devices. While such devices offer easy manufacture, sample collection and disposal, samples with only a few bacteria require time-consuming incubation. At Pittcon, Deiss will describe a a new mechanism he hopes can be implemented into elastomeric microfluidic devices to solve the challenge of detecting single bacterium without culture.

The use of microfluidic devices is also increasing among researchers using MS for proteomic analysis. Thermo Fisher Scientific can demonstrate how combining its mass spectrometers with 908 Devices’ ZipChip microfluidics separation technology delivers high-quality separation that revolutionizes MS biomolecule analysis.

In the field of clinical/biomedical diagnostics, the increasing need for portable integrated biosensors for point-of-care (POC) testing has led to the development of innovative, integrated and automated bioanalytical systems. Presenting at Pittcon, Charles Henry (Colorado State University) will discuss disease detection using paper-based analytical devices. Henry and team were the first to demonstrate (in 2009) how the integration of electrochemical detection and paper-based microfluidics provides an inexpensive and portable solution for POC monitoring. Also exhibiting at Pittcon will be Ametek Scientific Instruments, who provides single channel and multichannel potentiostats for the control of current during electroanalytical experiments.

German Gomez-Rios (University of Waterloo) will talk about the importance of developing new, high performance bioanalytical methods for drug development and POC diagnosis. Gomez-Rios will describe his team’s work on solid phase microextraction (SPME)-based devices for the extraction/enrichment of analytes from small sample volumes, that can be coupled with MS for rapid analysis. For the study, Thermo Fisher Scientific lent the team a triple quadrupole mass spectrometer, the TSQ-Quantiva. Waters also provide innovative triple quadrupole mass spectrometry, with the Xevo TQ range offering robust and reliable performance.

Facundo Fernandez (Georgia Institute of Technology) will discuss the use of ion mobility-mass spectrometry metabolomics for prostate cancer detection. Prostate-specific antigen (PSA) screening often results in overdiagnosis and overtreatment and rectal exams are limited by low detection rates in non-palpable growths. Now, Fernandez will describe high-throughput metabolic profiling of serum samples from prostate cancer patients using flow injection (FI) electrospray traveling wave ion mobility spectrometry (TWIMS) time-of-flight (ToF) MS. This enabled rapid cancer detection in serum samples with 90.2% sensitivity, 83.3% specificity and 87.4% accuracy.

Waters say their SNAPT G2-Si High Definition Mass Spectrometry system which is an ultra-performance liquid chromatography ion mobility time-of-flight mass spectrometry (UPLC-IM-TOF MS) system integrated with TWIMS provides the most complete characterization of complex mixtures and molecules.

Amanda Hummon (University of Notre Dame) will describe image-based techniques for evaluating drug treatments in 3D tumor models. In a 2016 paper, Hummon looked at the use of matrix-assisted laser desorption/ionization (MALDI) imaging mass spectrometry (IMS) for the evaluation of therapeutics in 3D cell culture systems. MALDI IMS enables in situ analysis of tissue sections for the acquisition of data on hundreds of unknown compounds in a single measurement, without prior knowledge of tissue composition. Hummon and team used MALDI IMS to study the distribution of a chemotherapy drug and found the approach was more effective at screening drug efficacy than 2D culture assays, as well as being a lower-cost, higher-throughput alternative to using animal models. Bruker provides MALDI-TOF systems that quickly and efficiently provide results in traditional MALDI applications as well as for cutting edge research in fields such as proteomics.

Lisa Jones (University of Maryland) will describe an in-cell protein footprinting method coupled with mass spectrometry for the study of protein structure. In a 2014 paper, Jones and team reported that multidimensional protein identification technology (MudPIT) improved hydroxyl radical footprinting for increased identification of quantifiable peptides. At Pittcon, Jones will discuss how the use of fast photochemical oxidation of proteins (FPOP) for the generation of hydroxy radicals in these footprinting experiments can be extended to enable the analysis of proteins inside cells, (in-cell FPOP), demonstrating the technique’s potential for studying proteins in their native cellular environment.

For a range of applications including protein identification and metabolite profiling, Thermo Scientific’s LTQ Orbitrap XL™ Hybrid Ion Trap-Orbitrap Mass Spectrometer serves as the ideal instrument. It features an HCD collision cell to enhance flexibility of fragmentation experiments for advanced proteomics and small molecule research.

Matthew Lockett (University of North Carolina) will discuss the tracking of cellular movement in tumor-like environments. Current cellular migration assays provide limited information on cellular responses to gradients, since they oversimplify the 3D environment of a tissue. In a 2015 paper, Lockett and team described a paper-based invasion assay that enables real-time monitoring of cellular movement with fluorescence microscopy. Using this technique, to study the invasion of cells cultured in the presence and absence of oxygen, Lockett and team were able to demonstrate that oxygen is a chemoattractant for more than one type of cancer cell.

Zeiss, a leading provider of microscopy solutions, will be exhibiting its high-performance microscopy products at the conference. The company’s portfolio features the LSM 700 laser scanning confocal microscope, which provides efficient separation of fluorescence signals where crosstalk is prevented and highly overlapping fluorophores are unmixed.

References


Chapter 3 – Bioanalytical Solutions in Biopharma

Numerous bioanalytical technologies are used as essential tools in many fields that shape our life today, ranging from the diagnosis of disease through to food safety and environmental conservation. Without such technologies, many of the products we use in everyday life would not have emerged and around one-fifth of Nobel prizes have been awarded to the creators of analytical theories, instruments or technologies in physics, chemistry, physiology and medicine.

The Japan Analytical Instruments Manufacturers Association (JAIMA) is a company headquartered in Chiyoda, Tokyo Japan that aims to progress science and technology by developing analytical technologies and encouraging progress in the analytical instruments industry. JAIMA is a leading contributor to the organization of events that will advance this field and therefore the economy, education, lifestyle and culture of Japanese citizens.

One of the aims of JAIMA is to harness the power of the internet of things (IoT) and big data analysis, for which sensing technology is crucial. JAIMA therefore expects analytical instrumentation will need to play a greater role in the detection and measurement of substance characteristics that can be translated into data for future use.

One aspect of improving analytical instrumentation is the development of high-throughput screening, a method that employs robotics, automation, high sensitivity detection, data processing software and sample handling devices to enable researchers to quickly perform millions of pharmaceutical, genetic or chemical tests.

At Pittcon, Atis Chakrabarti from Tosoh Bioscience LLC will discuss a high-throughput method for analyzing a variety of Immunoglobulin G subclasses from multiple sources including humans, rats, mice and rabbits. Using an analytical protein A affinity chromatography column, rapid separation and robust quantification of antibodies could be carried out, with analysis completed within two minutes. Pittcon exhibitor Thermo Scientific has an instrument portfolio that includes a range of analytical chromatography columns. Amongst them is the POROS™ Prepacked Protein A Affinity Column, which contains an immobilized recombinant Protein A functional group designed for high-throughput purification of antibodies.

In a 2009 article, Chakrabarti reported a successful high throughput method for the analysis of caffeine content in drinks. In 2004, all 25 countries in the EU started to require that any packaged drink containing more than 150 mg/L of caffeine be labelled as “high caffeine content,” leading to a need for a rapid and easy testing method for the quality control of caffeine. Chakrabarti’s article described a high-throughput reversed phase HPLC method that successfully achieved this purpose.
State-of-the-art reversed-phase and HPLC columns are available through Waters, who are also exhibiting at Pittcon.

Imaging technologies used in pharmaceutical applications enable researchers to gain powerful insights into the biological effects of drugs such as their distribution and binding ability. The imaging agents and technologies used in this field serve as important tools in the discovery of biomarkers, the study of patient outcomes in disease progression, and the development of new therapies. Pharmaceutical analyses provide information on the content, quality, stability and purity of pharmaceutical products. Metrohm, a leading provider of instruments and applications for drug quality control, monitoring and improvement, will be exhibiting at Pittcon this year. The company provides solutions for the analysis of active pharmaceutical ingredients (APIs), excipients and impurities, as well as starting materials and finished pharmaceutical products.

When a pharmaceutical is produced using biotechnology, it is referred to as a biopharmaceutical. These drugs are large, elegant and complex structures such as proteins or nucleic acids, rather than small molecules and they are designed and studied using 3D-modlieng systems. They perform their functions with high efficacy and limited side effects. Furthermore, treatment prototypes are continuously evolving and ongoing research is leading to new types of products.

Thermo Fisher Scientific’s instrument range and workflow solutions provide answers to simplifying biopharmaceutical analysis. The company supplies state-of-the-art technology for all aspects of protein characterization, from post translational modifications through to 3-D structures and protein aggregation.

However, the biopharma industry faces various challenges. Reliable reproduction of these large molecules at an industrial scale requires extremely sophisticated manufacturing capabilities and the genetically modified cells that are used to create the molecules need to be frozen for storing and thawed without becoming damaged. The biopharmaceutical also needs to be separated from the cells that were used to make them without their fragile structures being destroyed.

The current market value for biopharmaceuticals is now more than $150 billion and the range of therapeutics is larger than ever before. The market is also expected to explode over the next decade and Thermo Scientific believes there is a need to develop higher-throughput and more in -depth characterization methods.

At Pittcon, Gurmil Gendah (Shimadzu) will talk about using liquid chromatography mass spectrometry (LC-MS) platforms to increase the success of biotherapeutic process development and bioanalysis.

The cell culturing involved in the production of biopharmaceuticals requires routine monitoring of multiple medium conditions such as glucose, nitrogen and various other biologically important compounds. To meet the demands of this multi-component analysis, Gendah and team developed a high throughput LC/MS/MS method that can monitor the relative abundance of 95 compounds. In LC-MS, multiple components within mixtures can be separated by LC and then analyzed by MS to characterize those components with high specificity and detection. The high reliability, productivity and sensitivity of LC-MS make it an increasingly valuable and popular application in the world of pharma.

In a 2014 article, Gendah and colleagues also reported on a silica -based HPLC/UHPLC column (Thermo Scientific ™ GlycanPac ™ AXH -1) they developed for high-throughput and high-resolution separation and characterization of biologically relevant glycans from proteins using LC-MS. At the forefront of instrument sensitivity, stability and ease of operation is Pittcon exhibitor Shimadzu. One of the company’ liquid chromatograph mass spectrometers, the LCMS-8040, is equipped with newly improved ion optics and collision cell technology, for even higher sensitivity and enhanced monitoring.

Also presenting at Pittcon will be Amir Liba from Agilent, who will talk about the use of inorganic mass spectrometry for exploring new biopharmaceutical applications. A highlight of the talk will be how laser capture microdissection (ICP) MS can be used in new life science applications to analyze the uptake of therapeutic metal-containing drugs into single cells.

References


Chapter 4 – Combining Biosensors with Nanotechnology

Over recent years, there has been significant advancement in the development of nanotechnology. Many new nanomaterials have been created and researchers have been busy investigating their novel properties. One area that has seen great progress is the application of nanomaterials in biosensors. Nanomaterials-based biosensors represent a fusing of expertise in materials science, chemistry, biotechnology and molecular engineering for the enhancement of biological detection.

In particular, the unique physical, chemical, mechanical, magnetic and optical properties of gold nanoparticles, carbon nanotubes, magnetic nanoparticles and quantum dots has meant their application in biosensors has significantly improved the sensitivity of biomolecule detection. This technology could serve as a significant contributor to the enhancement of fields such as molecular recognition, disease diagnosis and environmental monitoring.

At Pittcon, Daniel Chiu (University of Washington) will discuss single molecule studies of nanoparticles. Chiu and team have developed microfluidic and nanofluidic systems that have enabled them to make new discoveries about single nanoparticles.

In a 2003 paper by Chiu, the development of techniques for the chemical analysis of nanometer-scale samples isolated from a single cell is discussed. The paper compares two techniques, namely capillary electrophoresis with laser-induced fluorescence (CE-LIF) matrix-assisted laser desorption/ ionization time of flight mass spectrometry (MALDI-TOF-MS). CE-LIF is a powerful analytical technique that can separate small sample volumes and where the use of LIF results in extremely low limits of detection. In MALDI-TOF-MS, molecules are ionized and a time of flight mass spectrometer is used to separate ions with identical kinetic energy based on their mass-to-charge ratio (m/z).

Chiu and team found that CE-LIF revealed the presence of many low molecular weight amino acids and said it may be an ideal technique for the chemical analysis of even the tiniest subcellular structures. Compared to CE-LIF, MALDI-TOF-MS demonstrated reduced sensitivity in detecting low molecular weight molecules. However, it did mainly reveal the presence of peptides and provides a suitable technique for the study of high molecular weight species. Exhibiting at this year’s Pittcon are leading scientific instrument manufacturers, Shimadzu and Bruker. Both company’s product portfolios include a range of outstanding and innovative solutions for quick and reliable MALDI-TOF analysis.

Michael Marty (University of Arizona) will discuss the advancement of nanodiscs as a platform for analyzing membrane proteins by mass spectrometry. The interactions between membrane proteins and lipids is often essential to their function, but their dynamic and heterogenous nature makes them challenging to define. The advancements Marty will describe open new doors for studying membrane proteins in nanoscale lipid bilayers. In a 2015 study, Marty and colleagues showed that nanodiscs can be used in native mass spectrometry to probe interactions between membrane proteins and the wider lipid environment.

Also exhibiting at Pittcon will be Wasatch Photonics, designer and manufacturer of world-class spectrometers and optical imaging equipment. Among the company’s portfolio is the new WP-series of spectrometers, high throughput instruments with an elegant opto-mechanical design.

Amemiya Shigeru (University of Pittsburgh) will discuss the use of scanning electrochemical microscopy (SECM) for high-resolution imaging of single biological nanopores. Shigeru’s work is focused on understanding chemically and biologically important membrane transport phenomena and the development of electrochemical sensors based on interfacial transport processes.
SECM is an electroanalytical scanning probe technique that can be used to image substrate topography and surface reactivity at high resolution.

Shigeru, will talk about how SECM imaging of individual nanopores in a physiological buffer shows that the permeability of a nanopore is related to its topography. Shigeru says a “plugged” nanopore is as permeable as an “open” nanopore, thereby indicating that the plug is a part of the transport or a highly permeable substance that is trapped in the pore.

Attendees interested in scanning electron microscopy (SEM) equipment, can enjoy exhibits by SEM solutions experts Hitachi and Zeiss. The ease of use, high quality imaging and compact design of Hitachi’s new generation tabletop microscopes redefine the capabilities of tabletop SEM and Zeiss offers a comprehensive portfolio of SEM instruments that provide high resolution surface imaging and excellent materials contrast.

Lane Baker will talk about the integration of dual-barrel membrane patch-ion channel probes (MP-ICPs) and scanning ion conductance microscopy (SICM) as a potentially revolutionizing spatially-resolved chemical sensing technique. Baker will explain a series of experiments that fully characterized the analytical performance of this new platform. SICM is a versatile scanning probe microscopy technique for use in nanoscale ion transport studies. It can measure electrochemical processes in a way that is complementary and sometimes superior to other electrochemical imaging methods.

Previous work by Baker has included the development of chemical and biochemical sensors that use synthetic conical nanopores to detect the presence of analytes. In a 2006 paper, Baker and colleagues describe a sensor that utilized molecular recognition elements bound to the nanopore mouth, blocking its tip, and thereby detecting the analyte. Another sensor made use of conical nanopores in a resistive-pulse type experiment, with the analyte detected via transient blockages in ionic current.

The rapid development of biosensor design is also highly relevant to the food industry, where foodborne pathogens are of increasing concern due to the recent worldwide spread of bacterial and microbial diseases resulting from agricultural trade. There is a growing demand for improved food safety, but conventional methods for detecting and identifying these microbial contaminants take several days to generate results. The advances in biosensor technologies have enabled more rapid detection of foodborne pathogens and are commonly used for the monitoring of food packaging and agricultural processes.

The International Association of Environmental Analytical Chemistry (IAEAC) is an organization that aims to provide scientists with opportunities to exchange cutting edge research in environmental analytical chemistry. The organization arranges many events including symposia, workshops and short course to serve as platforms for these exchanges. For example, the IAEAC is currently inviting researchers to the 40th International Conference on Environmental & Food Monitoring organised by the Institute for Food Analysis and Research (IIAA).

The Pittcon conference will also see presenters discuss the development of new bioanalytical technologies for point-of-care testing. John Connelly (Intellectual Ventures Laboratory) will discuss some of the technical and market barriers faced in creating and implementing impactful point-of-care diagnostics.

Jian-Hui Jiang (Hunan University) will discuss the use of nanobiotechnologies in the treatment of cancer. He will describe nanoscale assemblies of nucleic acids and peptides that enable efficient delivery of DNA or peptide probes into tumor cells. In a 2015 study, Jiang reported on the development of a novel electrostatic DNA nanoassembly that may serve as a valuable tool for low-abundance biomarker discovery and regulation in cell biology and theranostics.

Another of Jiang’s studies, published in 2013, describes the development of an immunoassay that provided sensitive detection of protein targets as a result of high surface-enhanced Raman spectroscopy (SERS) signal enhancement through controlled assembly of SERS nanoparticles. The SERS immunoassay platform may serve as a tool to aid accurate and early detection of disease biomarkers and to facilitate point-of-care diagnostics.

Also exhibiting at Pittcon will be manufacturer of photonics devices, Hamamatsu, which offers a surface-enhanced Raman spectroscopy (SERS) substrate that enhances the Raman scattering light from molecules, for highly sensitive Raman spectroscopic analysis.

References


Conclusion

At the core of all scientific disciplines dedicated to improving human health and the world we live in, is the use of analytical instrumentation.

Over recent years, advances in the development of these crucial technologies has revolutionized the analysis of living systems, the diagnosis of disease and the monitoring of treatment. The scope of bioanalytical techniques has expanded into high impact areas such as gene expression, metabolomics and proteomics, with further applications ranging from environmental, pharmaceutical and forensic analyses through to the monitoring of food safety.

With the increasing demands placed on researchers in the analysis of biomolecules, single living cells, proteomics and genomics, new bioanalytical platforms are emerging as invaluable tools in the advancement of many fields including biology, biomedicine, biomaterials science and engineering.

This year’s Pittcon will serve as a hub where experts in the field of analytical instrumentation can gather to hear all about the exciting new advances that have been made in this field, as well as those that are planned for the future.

The latest developments in analytical tools and techniques applying to a range of different technologies will be covered, including mass spectroscopy, mass spectrometry, chromatography, electroanalytical systems, micro- and nanofluidic devices and microscopy. The number of areas where these technologies can be applied is vast and include biomarker detection; point-of-care diagnostics; pesticide, toxin and contaminant screening in the food and agriculture industries; the evaluation and monitoring of therapeutics; drug quality control, and single nanoparticle studies.

Researchers have developed instrumentation that can perform at a higher-throughput and greater efficacy than ever before, but as realized by organizations such as The Japan Analytical Instruments Manufacturers Association (JAIMA) and The International Association of Environmental Analytical Chemistry (IAEAC), there is still a need to push for further development of analytical technologies and the analytical instruments industry.

World-leading manufacturers of state-of-the-art, analytical instrumentation are already signed up to attend Pittcon and will be showcasing the outstanding capabilities of their most recently developed technologies. Among the numerous major industry players attending are names such as Thermo Fisher Scientific Shimadzu, Waters, Bruker, BioRad, Ametek, Zeiss, Metrohm, Wasatch, Hamamatsu, Hitachi and Zeiss.

This year’s event will be host to a vast number of symposia where attendees can hear world-leading experts revealing the fascinating new discoveries they have made in areas ranging from biopharmaceutical characterization, oncology biomarker detection and the prevention of pathogenic outbreaks, through to the development of nanobiotechnologies for the treatment of cancer.

Any person involved in academia, research or industry that involves analytical instrumentation should not miss out on this informative and important event that is so relevant to our health and environment.