To start, could you provide a brief introduction to the Centre for Biospectroscopy at Monash University, outlining its key objectives and current research activities?

The Centre for Biospectroscopy, located in the School of Chemistry at Monash University, is dedicated to solving biomedical and biological problems using vibrational spectroscopic techniques. Vibrational spectroscopy is the study of how infrared light interacts with matter. All molecules vibrate at specifi c frequencies and a molecule will absorb an infrared photon if the frequency of that photon matches the frequency of the molecular vibration. We have a number of different research themes, which broadly fall under the banner of vibrational diagnostics and monitoring. Specifi c research areas include malaria diagnosis and treatment, cancer diagnosis, the testing of oocyte competence for in vitro fertilisation (IVF) programmes, stem cell research, heart disease, liver disease, phytoplankton studies, aquatic wetland studies and fundamental studies into how light interacts with cells and tissues.

What is your affi liation with the Centre?

Along with Professor Don McNaughton, Dr Phil Heraud and Professor John Beardall, I am

one of the founding members of the Centre for Biospectroscopy. It was established in 2002 when we held the inaugural Austral-Asian Biospectroscopy Conference at Suranaree University of Technology in Korat, Thailand. The conference, which attracted over 180 international scientists, introduced biospectroscopy into Southeast Asia and placed the Centre on the world stage. I am currently an Australian Research Council (ARC) Future Fellow and will take over the Directorship of the Centre following the retirement of Professor Don McNaughton.

Can you explain Raman acoustic levitation spectroscopy (RALS) and the importance of this research area?

RALS was a technique we applied to investigate cells and biofl uids in a containerless environment. This acoustic levitation device is comprised of a piezoelectric transducer (PET) and a refl ective sound plate. The PET generates an ultrasonic wave that is refl ected from the sound plate. The waves recombine and a number of standing wave nodes are generated. These nodes are very stable and samples can be placed into them and levitated, generally for several minutes, before becoming unstable or evaporating. There are several advantages to this approach. Firstly, the containerless environment results in no energy loss from the container walls, hence the signal-to-noise ratio is greatly improved. Secondly, it is a great way to concentrate fl uids and can be used to generate excellent crystals. Another advantage is that it allows the direct probing of the sample with another laser or UV source while simultaneously taking the measurement with the Raman spectrometer.

What have been your greatest challenges and successes to date?

Undoubtedly, our greatest challenge has been to take infrared technology into the

clinical environment. Our biggest success in terms of our cervical cancer studies was the development of a methodology to generate and diagnose early forms of cervical cancer using Fourier transform infrared spectroscopy (FTIR) mapping and imaging technology. The work was published in the Journal of Gynaecologic Oncology and has over 160 citations. Also, we were the fi rst group to record resonance Raman spectra of functional red blood cells. We applied the technology to monitor the oxygenation and deoxygenation of a single red blood cell where we assigned all of the individual bands. This then enabled us to look for applications in malaria diagnosis and sickle cell disease.

Lastly, where do you see your research progressing? Are there any projects you are currently involved in that you would like to highlight?

In collaboration with Dr Darren Creek (Monash Pharmacy College) and Dr Mark Tobin (Australian Synchrotron) we have been spectroscopically investigating the interactions of a new range of antimalarial drugs on living malaria parasites inside functional red blood cells. Using a specially designed sample holder we can record a transmission infrared spectrum through a single living cell. We have been invest igating the effects of drug incubation time on the infected cells and looking at how both infected and uninfected cells respond to these new drugs.

The ability to perform single cell analysis in a rapid time frame enables the acquisition of hundreds of spectra that are then processed using multivariate data and neural network analytical techniques. These pattern recognition techniques are designed to fi nd correlations in the data and enable us to determine the biochemical changes occurring in both the red blood cell and the parasite in response to the antimalarial drug.

Associate Professor Bayden Wood discusses how his research is forging new insights into biomedical and biological challenges through the application of cutting-edge vibrational spectroscopic techniques

An emerging fi eld

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ASSOCIATE PRO

FESSOR BAYD

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BIOSPECTROSCOPY, AN EMERGING field within the spectroscopic examination of living tissue or body fluids, combines information from the physical sciences with advanced computational analysis in order to shed new light on biological processes. This field is at the cutting-edge of biology research, continually developing knowledge about the structure and activity of biological molecules. Crucially, biospectroscopy has the potential to revolutionise clinical diagnostic processes, fulfilling the constant demand for new techniques that can identify diseases to a high level of objectivity, sensitivity and specificity.

Operating in this context, the Centre for Biospectroscopy within the School of Chemistry at Monash University has flourished over the course of the past decade. With its broad range of state-of-the-art equipment, strong roster of multidisciplinary staff and robust collaborations with other researchers, the world-class Centre is driving the development of bioscience and biotechnology. Associate Professor Bayden Wood, one of the Centre’s founding members, is an Australian Research Council (ARC) Future Fellow and a prominent scientist in the area of vibrational spectroscopy. Specifically, he is interested in using infrared and resonance Raman spectroscopy to monitor blood-borne diseases and in applying multivariate statistics and neural network architectures to the analysis of infrared and Raman spectra from bio-samples.

ADVANCING MALARIA DIAGNOSIS

The early, rapid diagnosis and treatment of malaria are essential for reducing mortality and preventing the overuse of antimalarial drugs. Existing techniques for malaria diagnosis include optical microscopy, rapid diagnostic tests (RDTs) and polymerase chain reaction (PCR) assays. However, they all suffer significant drawbacks. For example, microscopy depends on the presence of an experienced microscopist to analyse the sample while RDTs work by detecting HPR-2, which is only expressed by the parasite Plasmodium falciparum. This means that samples containing other malaria parasites could be falsely diagnosed as negative. Furthermore, the PCR assay – which is the most sensitive technique and the current gold standard – is very expensive and requires a high level of operational expertise.

In view of these limitations, Wood and his team have patented the use of a new diagnostic technique for malaria. Their rapid and inexpensive test uses infrared light to detect malaria at a very early stage of its development by identifying fatty acids produced by the parasite associated with the disease. Known as attenuated total reflection-Fourier transform infrared spectroscopy (ATR-FTIR), their technique uses small, portable and battery-operated technology, making it ideal for use in remote communities with poor accessibility. Importantly, its sensitivity matches that of the PCR assay, and it is able to quantify the number of infected cells as well as detecting different species of

Breakthroughs in biospectroscopyResearchers based in the Centre for Biospectroscopy at Monash University, Australia, are revolutionising the future of medical diagnosis and biological monitoring

ASSOCIATE PROFESSOR BAYDEN WOOD

48 INTERNATIONAL INNOVATION

BIOPECTROSCOPY – LIGHTING THE PATH TOWARDS THE FUTURE OF MEDICAL DIAGNOSIS AND BIOLOGICAL MONITORING

OBJECTIVE

To collaborate and deliver innovative biospectroscopy diagnostic knowledge advances for the rapid early detection of major diseases.

KEY COLLABORATORS

Associate Professor Matti Kiupel, University of Michigan, USA • Professor Max Diem, Northeastern University, USA • Professor Donald McNaughton; Dr Aazam Khoshmanesh; Professor Scott O’Neil; Dr Inaki Iturbe-Ormaetxe; Professor Brian Cooke; Professor John Beardall; Professor John Carol; Professor Glen Deacon; Dr Philip Heraud; Dr Darren Creek, Monash University, Australia • Professor Leann Tilley; Dr Matthew Dixon, The University of Melbourne, Australia • Dr Mark Tobin, Australian Synchrotron, Australia • Professor Małgorzata Baranska; Dr Kasia Marzec, Jagiellonian University, Poland • Professor Wojciech Kwiatek; Dr Ewelina Lipiec, The Henryk Niewodniczanski Institute of Nuclear Physics, Poland • Professor Patcharee Jearanaikoon, Khon Kaen University, Thailand

FUNDING

Australian Research Council (ARC) • National Health and Medical Research Council (NHMRC) • Agilent Technologies

CONTACT

Associate Professor Bayden Wood ARC Future Fellow

Monash University Department of Science School of Chemistry Wellington Road Clayton, Victoria 3800 Australia

T +61 3 9905 5721 E bayden.wood@monash.edu

bit.ly/1pgm8T6

BAYDEN WOOD is an ARC Future Fellow and a founding member of the Centre for Biospectroscopy at Monash University. He completed his PhD at Monash University in 1999 under the supervision of Professor Donald McNaughton, undertaking his first postdoctoral position at Hunter College, City University of New York, USA, with Professor Max Diem. He was awarded a Humboldt Fellowship in 2008 and undertook research at the Institute of Analytical Sciences in Dortmund with Associate Professor Volker Deckert and with Professor Dieter Naumann at the Robert Koch Institute in Berlin, Germany.

the malaria parasite. Additionally, it uses a computer algorithm to make the diagnosis. Currently, the one main disadvantage is that ATR-FTIR can only analyse one sample at a time, highlighting a need to develop a multi-cavity ATR-FTIR unit that can measure hundreds of samples simultaneously.

COMBATING CANCER

Cancer diagnosis is another major focus of research within the Centre. In fact, this was the area where Wood began his research, when as a student in 1995 he used FTIR to diagnose cervical cancer from exfoliated cells collected from routine Pap smears. In 2001, this initial research progressed into a collaborative project with Professor Max Diem from Northeastern University, USA, where the researchers explored tissue biopsies using FTIR mapping spectroscopy: “In this work we took maps of cervical tissue sections and processed them using a multivariate method called unsupervised hierarchical cluster analysis (UHCA),” Wood discloses. “This provided us with a way of correlating infrared data directly with conventional staining. The FTIR images are of such high quality that lymphocytes can now be classified distinctly from cancer cells as well as other cell types in the cervix.”

More recently, the researchers at the Centre have embarked on a collaborative project with Associate Professor Matti Kiupel from the University of Michigan, USA, where they are applying FTIR and Raman microscopy to explore canine cancer. Funded by an ARC Discovery grant, the researchers are attempting to identify infrared biomarkers that can distinguish between chemosensitive and chemoresistant canine lymphomas, thus helping to inform clinicians of the best treatment strategy. Additionally, in other cancer-related projects, Wood and his team are applying tip enhanced Raman scattering (TERS), a near-field technique that enables them to achieve a spatial resolution of approximately 10 nanometres. After successfully developing their own TERS system in Australia, the researchers have applied this technology to the study of how novel anticancer platinum-based drugs bind to DNA, from which they have developed atomic force microscopy (AFM) images that depict the process: “From these spectra we can determine the platinum binding sites and the effectiveness and specificity of the drug,” Wood explains.

DNA CONFORMATION

The Monash researchers have also applied their spectroscopy techniques to the study of DNA conformation. DNA conformation refers to the

different forms in which DNA exists; for example, in its hydrated state DNA has a more open form known as B-DNA while in its dehydrated state it exists in the narrower A-DNA form. Working in collaboration with one of his PhD students in 2011, Wood determined through the use of FTIR that this transition happens in all cells, even though DNA is tightly bound to histone proteins. The team discovered that for bacteria this transition may offer protection against desiccation.

Wood and his team found that the B-A-B DNA transition was also possible in bacteria, which are anucleated cells and, using ATR-FTIR, they investigated the hydrated, dehydrated and rehydrated states of bacteria. Importantly, they found that unlike in nucleated cells, bacterial cells become functional again after rehydration: “This could have potential applications in understanding bacterial resistance and explain how bacteria may be transferred from dry surfaces and rehydrate when hydrated in the human body,” Wood elucidates. “The challenge now is to use this knowledge to understand how bacterial DNA responds to other environmental changes like pH, cryogenetic freezing, oxidation, UV and other microbial agents.”

PROMISING POTENTIAL

Wood and his colleagues have made significant advances in the field of biospectroscopy to date. However, these advances are poised to become more pronounced in the near future, with spectroscopic technologies representing the next generation of point-of-care diagnostics. For instance, using ATR units and routine algorithms to analyse blood components will lead to the sensitive and rapid diagnoses of serious blood-borne diseases such as malaria and leukaemia. In addition, the use of focal plane array technology to analyse tissues should enable the generation of three-dimensional molecular images that will in turn establish surgical boundaries by highlighting where the tumour ends and begins. Finally, the portable nature of spectroscopic technologies could pave the way for the development of infrared detectors and infrared-tuned LEDs in mobile phones, with special apps enabling the diagnosis of disease. These exciting technological advances will reform future medical diagnoses and treatments, ultimately improving patient care and potentially saving many lives.

INTELLIGENCE

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