Category Archives: publication

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New microfluidic needle-like device developed

31 October:

A new paper from CNBP researchers (lead author Shilun Feng pictured) reports on the development of a microfluidic needle-like device that can extract and deliver nanoliter samples.

The paper, published in ‘Applied Physics Letters’ is accessible online.

Journal: Applied Physics Letters.

Publication title: A microfluidic needle for sampling and delivery of chemical signals by segmented flows.

Authors: Shilun Feng, Guozhen Liu, Lianmei Jiang, Yonggang Zhu, Ewa M. Goldys, and David W. Inglis.

Abstract: We have developed a microfluidic needle-like device that can extract and deliver nanoliter samples. The device consists of a T-junction to form segmented flows, parallel channels to and from the needle tip, and seven hydrophilic capillaries at the tip that form a phase-extraction region. The main microchannel is hydrophobic and carries segmented flows of water-in-oil. The hydrophilic capillaries transport the aqueous phase with a nearly zero pressure gradient but require a pressure gradient of 19 kPa for mineral oil to invade and flow through. Using this device, we demonstrate the delivery of nanoliter droplets and demonstrate sampling through the formation of droplets at the tip of our device. During sampling, we recorded the fluorescence intensities of the droplets formed at the tip while varying the concentration of dye outside the tip. We measured a chemical signal response time of approximately 3 s. The linear relationship between the recorded fluorescence intensity of samples and the external dye concentration (10–40 μg/ml) indicates that this device is capable of performing quantitative, real-time measurements of rapidly varying chemical signals.

 

Quantifying nanoparticles in colloidal suspension

26 October 2017:

A new paper featuring CNBP researchers (lead author Sandhya Clement pictured top left), demonstrates a simple, non-destructive method suitable for rapid evaluation of nanoparticles in colloidal suspension.

The paper, published in Nanotechnology is accessible online.

Journal: Nanotechnology.

Publication title: Quantification of nanoparticle concentration in colloidal suspensions by a non-destructive optical method.

Authors: Sandhya Clement, Brint Gardner, Wan Aizuddin W Razali, Victoria A Coleman, Åsa K Jämting, Heather J Catchpoole, Ewa M Goldys, Jan Herrmann and Andrei Zvyagin.

Abstract: The estimation of nanoparticle number concentration in colloidal suspensions is a prerequisite in many procedures, and in particular in multi-stage, low-yield reactions. Here, we describe a rapid, non-destructive method based on optical extinction and dynamic light scattering (DLS), which combines measurements using common bench-top instrumentation with a numerical algorithm to calculate the particle size distribution (PSD) and concentration. These quantities were derived from Mie theory applied to  measurements of the optical extinction spectrum of homogeneous, non-absorbing nanoparticles, and the relative PSD of a colloidal suspension. The work presents
an approach to account for PSDs achieved by DLS which, due to the underlying model, may not be representative of the true sample PSD. The presented approach estimates the absolute particle number concentration of samples with mono-, bi-modal and broad size distributions with <50% precision. This provides a convenient and practical solution for number concentration estimation required during many applications of colloidal nanomaterials.

New light source developed for fluorescent microscopy imaging

Aziz Rehman16 October 2017:

A tuneable and programmable integrating sphere light source for wide-field fluorescent microscopy imaging, employing nine light-emitting diodes (LEDs), has been successfully demonstrated and reported by CNBP researchers in a new paper (lead author Aziz ul Rehman pictured).

The paper, published in ‘Photodiagnosis and Photodynamic Therapy’ is accessible online.

Journal: Photodiagnosis and Photodynamic Therapy.

Publication title: Programmable LED-Based Integrating Sphere Light Source for Wide-Field Fluorescence Microscopy.

Authors: Aziz ul Rehman, Ayad G.Anwer, Ewa M.Goldys.

Abstract: Wide-field fluorescence microscopy commonly uses a mercury lamp, which has limited spectral capabilities. We designed and built a programmable integrating sphere light (PISL) source which consists of nine LEDs, light-collecting optics, a commercially available integrating sphere and a baffle. The PISL source is tuneable in the range 365–490 nm with a uniform spatial profile and a sufficient power at the objective to carry out spectral imaging. We retrofitted a standard fluorescence inverted microscope DM IRB (Leica) with a PISL source by mounting it together with a highly sensitive low- noise CMOS camera. The capabilities of the setup have been demonstrated by carrying out multispectral autofluorescence imaging of live BV2 cells.

Nanorubies for targeted bio-imaging

12 October 2017:

Researchers from the CNBP have released a new paper that examines the use of nanorubies for targeted bio-imaging activity. The work (lead author Varun Sreenivasan pictured) is trans-disciplinary in nature, drawing on the Centre’s collective knowledge in physics, pharmacology, chemistry, material science and embryology. The paper, published in ACS Applied Materials and Interfaces is accessible online.

Journal: ACS Applied Materials and Interfaces.

Publication title: Development of Bright and Biocompatible Nanoruby and its Application to Background-free Time-gated Imaging of G-protein Coupled Receptors.

Authors:  Varun K. A. Sreenivasan, Wan Aizuddin W Razali, Kai Zhang, Rashmi R Pillai, Avishkar Saini, Denitza Denkova, Marina Santiago, Hannah Brown, Jeremy Thompson, Mark Connor, Ewa M. Goldys, and Andrei V Zvyagin.

Abstract: At the forefront of development of fluorescent probes for biological imaging applications are enhancements aimed at increasing their brightness, contrast, and photostability, especially towards demanding applications of single molecule detection. In comparison with existing probes, nanorubies exhibit unlimited photostability and a long emission lifetime (3.7 ms), which enable continuous imaging at single-particle sensitivity in highly scattering and fluorescent biological specimens. However, their wide application as fluorescence probes has so far been hindered by the absence of facile methods for scaled-up high volume production and molecularly-specific targeting. The present work encompasses large scale production of colloidally stable nanoruby particles, demonstration of their biofunctionality and negligible cytotoxicity, as well as validation of its use for targeted biomolecular imaging. In addition, optical characteristics of nanorubies are found to be comparable or superior to state-of-the-art quantum dots. Protocols of reproducible and robust coupling of functional proteins to the nanoruby surface are also presented. As an example, NeutrAvidin-coupled nanoruby show excellent affinity and specificity to µ-opioid receptors in fixed and live cells, allowing wide-field imaging of G-protein coupled receptors with single particle sensitivity.

Nano ‘terminator style’ antennae

12 October 2017:

The liquid metal, shape-shifting T-1000 Terminator cyborg, featuring in a 1991 science-fiction film Terminator 2, was made possible due to breakthroughs in computer-generated imagery.

Some 25 years later, using breakthroughs in physics and chemistry CNBP scientists Dr Ivan Maksymov and Prof Andy Greentree at RMIT University have shown reconfigurable liquid-metal optical nanoantennae.

“An optical nanoantenna operates similarly to a conventional radio-frequency antenna, but its size is millions of times smaller” explains Dr Ivan Maksymov, “so it can receive and emit light similar to how a mobile phone antenna receives and emits radio waves.”

“The shape and length of the metal components that form a radio-frequency antenna determine its major properties such as operating frequency and radiation pattern,” explains Prof Andy Greentree, “so a liquid metal that can change its shape by applying voltage allows for changing antenna properties, which otherwise is difficult to achieve with fixed metal parts.”

“However, reconfigurability of optical nanoantennae is even more difficult to achieve than in radio-frequency antennae, because of their small size and lack of technologies enabling us to apply voltage to nanoscale sized objects. Therefore, we proposed a new solution – reconfiguration of liquid-metal nanoparticles using ultrasound.”

Continued Dr Maksymov, “A liquid-metal nanoparticle can change its shape due to capillary oscillations, which can be seen by everybody when observing water drops falling from a leaking kitchen tap. Drops change their shape when they detach from the tap and fall into the sink. In our work, we use ultrasound to change the shape of liquid-metal nanodroplets, which changes the nanoantenna’s operating frequency.”

“But fundamental physics remains the same as in the case of water drops.”

The paper ‘Dynamically reconfigurable plasmon resonances enabled by capillary oscillations of liquid-metal nanodroplets’ is accessible online.

Lung tissue aspiration

11 October 2017:

A new publication from CNBP researchers explores the integration of an optical coherence tomography (OCT) probe into a flexible needle for lung tissue aspiration. The paper (lead author Jiawen Li pictured), was published in the Journal of Biomedical Optics and is accessible online.

Journal: Journal of Biomedical Optics.

Publication title: Flexible needle with integrated optical coherence tomography probe for imaging during transbronchial tissue aspiration.

Authors: Jiawen Li; Bryden C. Quirk; Peter B. Noble; Rodney W. Kirk; David D. Sampson; Robert A. McLaughlin.

Abstract: Transbronchial needle aspiration (TBNA) of small lesions or lymph nodes in the lung may result in nondiagnostic tissue samples. We demonstrate the integration of an optical coherence tomography (OCT) probe into a 19-gauge flexible needle for lung tissue aspiration. This probe allows simultaneous visualization and aspiration of the tissue. By eliminating the need for insertion and withdrawal of a separate imaging probe, this integrated design minimizes the risk of dislodging the needle from the lesion prior to aspiration and may facilitate more accurate placement of the needle. Results from in situ imaging in a sheep lung show clear distinction between solid tissue and two typical constituents of nondiagnostic samples (adipose and lung parenchyma). Clinical translation of this OCT-guided aspiration needle holds promise for improving the diagnostic yield of TBNA.

New med-tech zinc sensor developed

27 September 2017:

A new zinc sensor has been developed and reported by CNBP researchers, which will allow for a deeper understanding of the dynamic roles that metal ions play in regulating health and disease in the living body.

The research, published in the journal ‘ACS Omega’ reports that the newly designed chemical sensor can detect and measure zinc levels in cells. It also has the functionality and portability to take continuous or repeated measurements within a single biological sample.

“This makes the sensor potentially suitable for use in future diagnostic tools that could open up entirely new windows into the body,” says lead author of the research Dr Sabrina Heng (pictured), Research Fellow at the ARC Centre of Excellence for Nanoscale BioPhotonics (CNBP), at the University of Adelaide.

Read the full CNBP media release here and the publication here.

Journal: ACS Omega.

Publication title: A Rationally Designed Probe for Reversible Sensing of Zinc and Application in Endothelial Cells.

Authors: Sabrina Heng, Philipp Reineck, Achini K. Vidanapathirana, Benjamin J. Pullen, Daniel W. Drumm, Lesley J. Ritter, Nisha Schwarz, Claudine S. Bonder, Peter J. Psaltis, Jeremy G. Thompson, Brant C. Gibson , Stephen J. Nicholls, and Andrew D. Abell.

Abstract: Biologically compatible fluorescent ion sensors, particularly those that are reversible, represent a key tool for answering a range of fundamental biological questions. We report a rationally designed probe with a 6′-fluoro spiropyran scaffold (5) for the reversible sensing of zinc (Zn2+) in cells. The 6′-fluoro substituent overcomes several limitations normally associated with spiropyran-based sensors to provide an improved signal-to-background ratio and faster photoswitching times in aqueous solution. In vitro studies were performed with 5 and the 6′-nitro analogues (6) in HEK 293 and endothelial cells. The new spiropyran (5) can detect exogenous Zn2+ inside both cell types and without affecting the proliferation of endothelial cells. Studies were also performed on dying HEK 293 cells, with results demonstrating the ability of the key compound to detect endogenous Zn2+ efflux from cells undergoing apoptosis. Biocompatibility and photoswitching of 5 were demonstrated within endothelial cells but not with 6, suggesting the future applicability of sensor 5 to study intracellular Zn2+ efflux in these systems.

New technique to aid bladder cancer diagnosis

25 September 2017:

A new and innovative automated computer technique has been developed by CNBP researchers that is able to significantly aid in the diagnosis of bladder cancer.

The technique—which allows suspect lesion images to be quickly and effectively analysed and then classified for cancer risk, has been reported in the medical journal ‘Urologic Oncology’.

“What we’ve done is develop a computer program to carry out an automated analysis of cystoscopy images,” says lead author of the research, Dr Martin Gosnell, Researcher at the ARC Centre of Excellence for Nanoscale BioPhotonics (CNBP) at Macquarie University and Director at Quantitative Pty Ltd.

Cystoscopy is one of the most reliable methods for diagnosing bladder cancer explains Dr Gosnell.

“Images are taken of the bladder and its insides for suspicious lesions during a routine clinical patient evaluation. Dependent on the findings, this initial scan can then be followed up by a referral to a more experienced urologist, and a biopsy of the suspicious tissue can be undertaken.”

The issue says Dr Gosnell is that the clinician examining the initial images makes a visual judgement based on their professional expertise as to the next steps of action that should be undertaken—such as the need to take a biopsy for subsequent pathological analysis.

“Potential errors and unnecessary further interventions may result from the subjective character of this initial visual assessment.”

“What we’ve done,” says Dr Gosnell, “is to create an automated image analysis technique which can identify tissue and lesions as either high-risk or minimal-risk.”

Read the full CNBP media release and the science paper here.

Journal: Urologic Oncology.

Publication title: Computer-assisted cystoscopy diagnosis of bladder cancer.

Authors: Martin E. Gosnell (pictured top), Dmitry M. Polikarpov, Ewa M. Goldys, Andrei V. Zvyagin and David A. Gillatt.

Abstract:

Objectives

One of the most reliable methods for diagnosing bladder cancer is cystoscopy. Depending on the findings, this may be followed by a referral to a more experienced urologist or a biopsy and histological analysis of suspicious lesion. In this work, we explore whether computer-assisted triage of cystoscopy findings can identify low-risk lesions and reduce the number of referrals or biopsies, associated complications, and costs, although reducing subjectivity of the procedure and indicating when the risk of a lesion being malignant is minimal.

Materials and methods

Cystoscopy images taken during routine clinical patient evaluation and supported by biopsy were interpreted by an expert clinician. They were further subjected to an automated image analysis developed to best capture cancer characteristics. The images were transformed and divided into segments, using a specialised color segmentation system. After the selection of a set of highly informative features, the segments were separated into 4 classes: healthy, veins, inflammation, and cancerous. The images were then classified as healthy and diseased, using a linear discriminant, the naïve Bayes, and the quadratic linear classifiers. Performance of the classifiers was measured by using receiver operation characteristic curves.

Results

The classification system developed here, with the quadratic classifier, yielded 50% false-positive rate and zero false-negative rate, which means, that no malignant lesions would be missed by this classifier.

Conclusions

Based on criteria used for assessment of cystoscopy images by medical specialists and features that human visual system is less sensitive to, we developed a computer program that carries out automated analysis of cystoscopy images. Our program could be used as a triage to identify patients who do not require referral or further testing.

Below: Dr Martin Gosnell and Prof Ewa Goldys.

First reversible ‘turn-off’ sensor for Glutathione

6 September 2017:

The first reversible ‘turn-off’ sensor for Glutathione has been reported by CNBP researchers in a paper published in the science journal Biosensors.

The paper is accessible online (open access).

Dr Sabrina Heng notes:

γ-Glutamyl-cysteinyl-glycine (GSH) plays a critical role in maintaining redox homeostasis in biological systems and a decrease in its cellular levels is associated with disease. Many diseases including Parkinson’s, cancer, heart diseases and Alzheimer’s are indicated by a decrease in GSH levels. In this case, a ‘turn on’ sensor would result in reduced fluorescence relative to healthy cells. An important advance would come from the development of a sensor that is measurably turned off by GSH and back on by a lower level of GSH. This would then provide an opportunity to sense reduced GSH levels during the onset of important diseases.

With that in mind we have rationally designed, to the best of our knowledge, the first reversible, reaction-based ‘turn-off’ probe that is suitable for sensing decreasing levels of GSH, a situation known to occur at the onset of various diseases.  We have demonstrated that the sensor can be used to detect changes of intracellular GSH in live HEK 293 cells to provide a potentially regenerable sensor for monitoring lower levels of intracellular GSH as associated with the onset of important diseases.

Journal: Biosensors.

Publication title: A Rationally Designed Reversible ‘Turn-Off’ Sensor for Glutathione.

Authors: Sabrina Heng (pictured), Xiaozhou Zhang, Jinxin Pei and Andrew D. Abell.

Abstract: γ-Glutamyl-cysteinyl-glycine (GSH) plays a critical role in maintaining redox homeostasis in biological systems and a decrease in its cellular levels is associated with diseases. Existing fluorescence-based chemosensors for GSH acts as irreversible reaction-based probes that exhibit a maximum fluorescence (‘turn-on’) once the reaction is complete, regardless of the actual concentration of GSH. A reversible, reaction-based ‘turn-off’ probe (1) is reported here to sense the decreasing levels of GSH, a situation known to occur at the onset of various diseases. The more fluorescent merocyanine (MC) isomer of 1 exists in aqueous solution and this reacts with GSH to induce formation of the ring-closed spiropyran (SP) isomer, with a measurable decrease in absorbance and fluorescence (‘turn-off’). Sensor 1 has good aqueous solubility and shows an excellent selectivity for GSH over other biologically relevant metal ions and aminothiol analytes. The sensor permeates HEK 293 cells and an increase in fluorescence is observed on adding buthionine sulfoximine, an inhibitor of GSH synthesis.

Charge transfer in helical peptides

1 September 2017:

Understanding the electronic properties inherent to peptides is crucial for controlling charge transfer, and precursory to the design and fabrication of bio-inspired next generation electronic components.

However, to achieve this objective one must first be able to predict and control the associated charge transfer mechanisms.

Here CNBP researchers demonstrate for the first time a controllable mechanistic transition in peptides resulting directly from the introduction of a side-bridge.

Journal: RSC Advances.

Publication title: A controllable mechanistic transition of charge transfer in helical peptides: from hopping to superexchange.

Authors: Jingxian Yu (pictured), John R. Horsley and Andrew D. Abell.

For more information, access the paper here.