Flow Cytometry Core Facility

Flow Cytometry Core Facility

The ONJCRI Flow Cytometry Core Facility is a shared resource laboratory which provides state-of-the-art analytical cytometry and high speed cell sorting services to the research community of our Institute, affiliates, and external users. Our goal is to provide high quality service to researchers and proactively introduce flow cytometric methods into new research areas. An important part of our mission is to teach this technology to students, staff, and investigators.

Cell sorter is operated by facility staff from 10:00am-5:30pm Wednesday to Friday. Extended sorting hours are available upon request. Self-service sorting is also available for trained researchers. Analysers available 24 hours / 7 days a week for self-service use (for suitably trained users only).

Services and Instrumentation

Our Flow Cytometry Core Facility provides:

  • Operator assisted Cell Sorting
  • Independent acquisition and analysis (users must go through the training process first)
  • Operator assisted acquisition and analysis
  • Consultation and assistance with experimental design and data analysis

Currently we house four instruments:

  1. BD FACSARIA III is a 4 laser, 12 detector cell sorter capable of measuring 10 colours simultaneously plus cell size and complexity parameter. The lasers in this instrument are 488nm (Blue), 633nm (Red), 561nm (Yellow-Green) and 405nm (Violet). It can sort up to 4 populations simultaneously into different size tubes, tissue culture plates, and onto individual slides. Sorting on the Aria can be performed using high-pressure (70 µm), intermediate-pressure (85 µm), or low-pressure (100 µm) nozzle. Instrument operates on a PC-based platform using FACSDiva (version 8) software.
  2. 2x BD FACSCanto II is three lasers, 10 detector analysis flow cytometer, capable of measuring 8 colors simultaneously plus cell size and complexity parameter. The lasers in this instrument are 488nm (Blue), 633nm (Red) and 405nm (Violet). It also features a High Throughput Sampler (96 well Plate Loader). These instruments operate on a PC-based platform using FACSDiva (version 8) software.
  3. AutoMACS Pro Separatoris a benchtop instrument for high-speed magnetic cell sorting of multiple samples. Employing MACS® Technology, it is designed for cell isolation in a fully automated, walk-away fashion. It can be used as a pre-sorter for speeding up flow sorting.
  4. Data Analysis Software – FlowJo– is a comprehensive analysis package specifically designed for handling list mode data generated by cytometers. ONJCRI provides several Flow Jo dongles, which are available to Institute members at no cost. Users can download the latest FlowJo version for Windows or Mac by contacting Institute’s IT Department.

Contact

If you would like to discuss opportunities to use our platform technologies and services, please contact:

Mark Frewin
Laboratory and Facilities Manager
P +61 3 9496 5299
M +61 434 242 518

Email

Mammalian Protein Expression Facility

Mammalian Protein Expression Facility

This unique facility is located in a dedicated suite of cleanrooms and can produce small to large amounts of high quality recombinant proteins and antibodies for use in medical research.

Cell Line Development
Transient expression, stable expression, and isolation/enrichment of high producing clones can be performed.

Production Systems
A range of production systems are available depending on the scale of production required. Perfusion systems are able to produce material every two days ranging from tens of milligrams to several hundreds of milligrams per harvest. A proprietary biphasic approach to production is used during protein production in stirred tank bioreactors. Yields up to 4g/L of mAb in a GS CHO cell line can be achieved.

Purification and Product Characterisation
Production material is purified using a variety of chromatography techniques such as affinity, size exclusion and ion exchange. Operating procedures are in place to allow for flexibility to process materials ranging from milligrams to several grams of material.

Quality assessments are conducted through a combination of SDS-PAGE, size exclusion chromatography using HPLC, and binding kinetics using BIAcore.

Contact

If you would like to discuss opportunities to use our platform technologies and services, please contact:

Mark Frewin
Laboratory and Facilities Manager
P +61 3 9496 5299
M +61 434 242 518

Email

VECTRA Multi-Spectral Imaging Platform

VECTRA Multi-Spectral Imaging Platform

The Vectra multi-spectral immuno-histochemistry platform is the first of its kind in Australia. It allows researchers to more accurately define the immune microenvironment, and helps detect mechanisms a tumour may use to evade the immune system.Its acquisition was supported by a grant from the Ian Potter Foundation in collaboration with La Trobe University and the Peter MacCallum Cancer Centre.

The Vectra platform and its analysis software allows researchers to quantify cells which are positive for one or both of the molecules and to analyse their localisation within the cells (for example, in the membrane, cytoplasma or nucleus).

Contact

If you would like to discuss opportunities to use our platform technologies and services, please contact:

Mark Frewin
Laboratory and Facilities Manager
P +61 3 9496 5299
M +61 434 242 518

Email

Functionalities

The Vectra platform can simultaneously detect up to seven different proteins of interest on one FFPE-tissue slide using Opal chemistry and spectral un-mixing.

It has an integrated automated slide-loader for up to 200 slides, which can be loaded and scanned, fully automated by user-defined protocols. Pre-scans of whole tissue slides will be performed at 4x or 10x magnification (RGB) followed by high-power-field (HPF) imaging of regions of interest in the multispectral mode (20x or 40x magnification).

The multiplex IHC images lead to a comprehensive understanding of complex cellular interactions which is not accessible by other methods. Opal follows the standard IHC workflow using unlabelled primary antibodies, followed by the addition of anti-species-HRP conjugate and detection substrate. Opal fluorescent detection substrates bind covalently near the epitope, allowing subsequent removal of antibodies to clear the tissue for detection of the next target. The signal remains intact after antibody removal.

Localisation studies of proteins in cellular compartments, co-localisation of proteins and quantification of cells expressing single markers or marker combinations can be subsequently performed with specialised software available on several work stations at the ONJCRI.

Spatial relationships of cell types to each other and within the tissue context can be analysed using Spotfire software. Scanning of the slides and selection of tissue sections and areas of interest can be automated on the Vectra system and allows for review by a pathologist before analysis. Due to the TSA based signal amplification, primary antibody concentration can be reduced up to 100X and all markers of interest can be detected with antibodies raised in the same species.

Aperio AT2 Whole Slide Scanner

The Aperio AT2 is a high-volume microscope capable of digitalising histochemical stained slides. The Aperio AT2 has a capacity for up to 400 slides that can be scanned with a sustained rate of 50 slides per hour at 20 X. With high first scan success rate, your images will be uploaded to our microscopy server, allowing remote access for collaborative research teams worldwide.

Analysis can be performed using the Aperio ImageScope – Pathology Slide Viewing Software. This software contains integrated macros and algorithms for the analysis of many common pipelines for investigating your histochemical stained tissues. Image export is easily achieved, allowing free access to your datasets in your analysis software of choice.

BOND RX Fully Automated Research Stainer

The BondRX allows you to fully automate IHC, ISH, FISH, CTC and multiplexing staining experiments. Reduce your manual work to increase your efficiency and consistence with your staining experiments. The BondRX allows you to customise all steps in your staining protocol from baking and dewaxing, antigen retrieval to the length and temperature of your staining approach. The BondRX has a 30-slide capacity, with finished trays of 10 slides being capable of replaced continuously.  Up to three separate staining protocols can be run simultaneously.

The BondRX allows you to leverage established protocols and reagents or develop completely custom and reproducible staining experiments with your own novel agents. The BondRX dramatically reduces the time required to perform your experiment. A bench run seven-plex experiment would require approximately 5 days of hands-on time manual time but with the BondRX this is reduced to a reproducible and consistent ~17 hours.


Prof Jonathan Cebon

Prof Jonathan Cebon

Head, Cancer Immunobiology Program

As a physician-scientist, every day I have the opportunity to leverage the partnership between scientific research and care to improve outcomes for cancer patients.

My experience spans two decades of translating novel immunotherapies into clinical application for the treatment of melanoma. I have successfully integrated laboratory and clinical teams to better understand immune responses to cancer and develop breakthrough therapies based on this knowledge. More recently, we have learned that it is critical to understand the biology of cancer plasticity to identify the mechanisms that allow tumours to resist or escape immune control. My research brings these elements together with the intention of achieving greater clinical benefit. Across 20 years, I have published more than 150 clinical and translational publications with a focus on the cell biology of melanoma and immunology and patented eight novel therapies. Finding the cure for cancer is dependent on the collaboration of scientific minds across generations. I am actively involved in the training and mentoring of postgraduate research students, post doctorals, Clinical Research Fellows, and Oncology Advanced Trainees.



The ACRF Centre for Translational Cancer Therapeutics and Imaging

The ACRF Centre for Translational Cancer Therapeutics and Imaging

This Centre was established for medical research and preclinical investigations and houses a range of advanced imaging technologies and platforms including:

Contact

If you would like to discuss opportunities to use our platform technologies and services, please contact:

Mark Frewin
Laboratory and Facilities Manager
P +61 3 9496 5299
M +61 434 242 518

Email

Take a look inside

PET MRI imaging

By combining a high-performance PET system and compact MRI technology, NanoScan® PM PET/MRI provides preclinical whole body soft tissue images with detailed quantitative imaging data within just one study. The PET camera offers quantitative 3D spatial resolution at 700 µm combined with a uniquely large field-of-view. The 1 Tesla permanent magnet for MRI provides 100 µm resolution with advanced sequences and ensures robust imaging across a broad range of biological applications including:

  • Oncology
  • Tumour biology
  • Stem cell investigations
  • Regenerative medicine
  • Neuroscience and receptor studies
  • Cardiology
  • Immunology and inflammation
  • Multimodal contrast agent development
  • Animal model development and phenotyping
  • Nephrology
  • Pharmacokinetics
  • PET development of radiotracers

SPECT CT imaging

The NanoSPECT/CTTM is an in-vivo molecular imaging system suitable for use with small animals and unifies functional (SPECT) and anatomical (CT) imaging procedures for preclinical investigations. With 250μm 3D SPECT and 30μm CT spatial resolution, exceptional quantification is permitted with an accuracy over 97%.

The system enables the examination of bio-chemical processes in healthy and disease models (including cancer, diabetes and stroke). It determines the localisation of radio-labelled compounds used as probes for the disease state, and monitors the efficacy of interventions or therapies. In living subjects, the effect of drugs in development can be monitored and compared in real time and at multiple time points within the one subject. Drug localisation as well as uptake can be quantified and visualised from the data collected.  The system is also suitable for monitoring genetic modifications and gene-therapeutic healing procedures using appropriate preclinical models.

IVIS spectrum imaging

The IVIS® Spectrum is a versatile and advanced in vivo imaging system, which uses a novel patented optical imaging technology to allow non-invasive longitudinal monitoring of disease progression, cell trafficking and gene expression patterns in living animals. High efficiency filters and spectral un-mixing algorithms take full advantage of bioluminescent and fluorescent reporters across blue to near-infrared wavelengths. It also offers single-view 3D tomography for both fluorescent and bioluminescent reporters, which can be analysed in an anatomical context using a Digital Mouse Atlas or registered with the IVIS multimodality module to other tomographic technologies such as MR, CT or PET.

For advanced fluorescence pre-clinical imaging, the IVIS Spectrum can use either trans-illumination (from the bottom) or epi-illumination (from the top) to illuminate in vivo fluorescent sources. 3D diffuse fluorescence tomography can be used to determine source localisation and concentration using the combination of structured light and trans illumination fluorescent images. The instrument is equipped with 10 narrow band excitation filters (30nm bandwidth) and 18 narrow band emission filters (20nm bandwidth), which assist in significantly reducing autofluorescence, via the spectral scanning of filters and the use of spectral unmixing algorithms. In addition, the spectral unmixing tools allow the researcher to separate signals from multiple fluorescent reporters in the same animal.


Research Platforms

Our research services

All research activities at the Olivia Newton-John Cancer Research Institute are enhanced and supported by outstanding platform technologies, facilities and technical expertise.

We also have a number of platform technologies and services which can be utilised by other Institutions and organisations. These include:

Contact

If you would like to discuss opportunities to use our platform technologies and services, please contact:

Mark Frewin
Laboratory and Facilities Manager
P +61 3 9496 5299
M +61 434 242 518

Email

Receptor Biology Laboratory

Our research focus

Eph receptors

Eph receptors are cell surface proteins that guide cell migration by binding to other cell-bound proteins (ephrins) on adjacent cells, thereby controlling cell-cell adhesion. Ephs coordinate cell movement during normal development of tissue and organ boundaries, and the vascular and neural networks. They are generally scarce in adults but reappear in cancers, where they are often on early ‘progenitor’ cell types, associated with blood vessel formation, and tumour cell invasion and spread.

EphA3 is a particular focus, which we investigate in tumour models, using ‘knock-down’ mice or by treatment with a specific antibody we helped develop, with the aid of drug payloads to specifically target the tumour microenvironment.

ADAM metalloproteases

ADAM metalloproteases (or ADAMs) are cell membrane-bound proteases that shed a range of other membrane proteins, regulating the activity of diverse cell surface receptors. These include Ephs and other receptors controlling cancer cell growth, drug resistance, and invasion and spread to other tissues. ADAMs also play an important role in the tumour microenvironment and in inflammation.

ADAM10 and 17 are of particular interest and we are investigating their function in tumours, as well as developing antibodies and antibody-drug conjugates as potential new therapies.

Fast facts

Cell surface (or membrane) receptors are proteins attached to a cell’s exterior which can receive external signals, usually by binding with another protein. The bound receptors then send signals into the cell to modify its behaviour, including movement, proliferation and survival.

They are distinctive in that they bind to proteins attached to adjacent cells. This allows them to control cell adhesion, migration and invasion. They are important in normal embryonic development but reappear in certain cell types in tumours and their surrounding environment, including new tumour blood vessels, which support tumour growth and spread.

A protease is a protein which cuts other proteins in a very controlled manner. ADAMs are a type of cell surface metalloprotease (‘metallo’ refers to their dependence on metal ions). ADAM10 and 17 control the activity of various cell surface receptors and are essential in normal cellular development. However they become overly active in tumours and their surrounding environment by supporting tumour growth, survival and drug resistance.

Recent publications

Journal of Experimental Medicine

An activated form of ADAM10 is tumor selective and regulates cancer stem-like cells and tumor growth.
DOI: 10.1084/jem.20151095

View abstract
Cancer Research

Targeting EphA3 inhibits cancer growth by disrupting the tumor stromal microenvironment.

DOI: 10.1158/0008-5472.CAN-14-0218

View abstract
Cell

Adam meets Eph: an ADAM substrate recognition module acts as a molecular switch for ephrin cleavage in trans.

DOI: 10.1016/j.cell.2005.08.014

View abstract

Our team

Meet our researchers


Tumour Targeting Laboratory

Our research focus

Targeting Strategies in Cancer

We have identified a series of molecules selectively expressed on cancer cells, and in the tumour microenvironment, that can be targeted for cancer therapy. This includes conformationally exposed receptor epitopes, such as found on the Epidermal Growth Factor Receptor (EGFR) and which led to the development of mAb806 and our first-in-human trials of this molecule. Our research findings have provided a new paradigm in antibody-based targeting and therapy of solid tumours. This work has expanded to incorporate structure-function studies of additional novel antibodies we have developed that target cell surface receptors and tumour microenvironment in tumours, as well as investigating mechanisms of resistance to antibody therapeutics, and targeting key molecules involved in sustaining the tumour microenvironment.

Antibody Engineering

We have developed techniques for generation and humanisation of antibodies. Recent molecular engineering, structural and modelling approaches in our laboratory have defined novel Fc:FcRn and Fc:FcγR interactions, which result in improved immune effector function and bioavailability of humanised antibodies. We have also developed strategies to deliver payloads specifically to tumours through conjugation of drugs, toxins and isotopes to recombinant antibodies, peptides and nanoparticles, both in preclinical models and more recently in clinical trials in cancer patients. These studies are showing encouraging results in patients with cancers of the brain, colon and breast, as well as other solid tumours.

Tumour Payload Delivery

The development of recombinant antibodies for cancer therapy has emerged as one of the most promising areas in oncology therapeutics, both as single agents, and for payload delivery. The concept of being able to deliver toxins through antibody-drug conjugates, or radiotherapy by antibody-radioisotope conjugates targeting the payload to sites of disease, are exciting and promising approaches that we are exploring preclinically and clinically with antibodies developed in our laboratory.

Novel Metabolic Tracers

An exciting recent development in the molecular imaging of cancer comes from the identification of critical biochemical pathways responsible for tumour growth and metastasis, and immune targets which can be exploited for therapy, which can be imaged with novel SPECT and positron emission tomography (PET) tracers. Taking the discovery of novel metabolic tracers in the laboratory to clinical trials is a major focus of our molecular imaging / PET research program and is leading to a deeper understanding of tumour biology and therapy response.

Fast facts

An immune protein normally produced within the body, that can recognise and eliminate foreign substances that can cause tissue damage.

Cancer scientists are able to develop and introduce antibodies into the body. They can be designed to recognise and target a specific feature of the tumour to inhibit or stop tumour growth.

Molecular imaging technologies which allow researchers to see whether treatments are effectively targeting a tumour, and how a tumour responds to treatment.

Recent publications

Molecular Cancer Therapeutics

Characterization of ABT-806, a Humanized Tumor-Specific Anti-EGFR Monoclonal Antibody.

DOI: 10.1158/1535-7163.MCT-14-0820

View abstract
Journal of Clinical Oncology 

Phase I Imaging and Pharmacodynamic Trial of CS-1008 in Patients With Metastatic Colorectal Cancer.

DOI: 10.1200/JCO.2014.60.4256

View abstract
Nature Medicine

Microenvironmental control of breast cancer subtype elicited through paracrine platelet-derived growth factor-CC signaling.

DOI: 10.1038/nm.4494

View abstract

Our team

Meet our researchers

  • Prof Andrew Scott AM - Head, Tumour Targeting Program | Head, Tumour Targeting Laboratory | Co-Director, Centre for Research Excellence in Brain Cancer | Director, Department Of Molecular Imaging And Therapy, Austin Health   Publications
  • Hui Gan - Clinical Research Lead | Clinician Scientist | Co-Director, Centre for Research Excellence in Brain Cancer | Director, Cancer Clinical Trials Centre, Austin Health  Publications
  • Zhanqi Liu - Associate Investigator
  • Ingrid Burvenich - Postdoctoral Research Fellow Publications
  • Christian Wichmann - Postdoctoral Research Fellow Publications
  • Alexander Mcdonald - Postdoctoral Research Fellow Publications
  • Sagun Parakh - Postdoctoral Research Fellow Publications
  • Benjamin Gloria - Senior Production Scientist Publications
  • Laura Allan - Senior Research Officer
  • Diana Cao - Senior Research Officer
  • Nancy Guo - Senior Research Officer Publications
  • Nhi Huynh - Senior Research Officer
  • Angela Rigopoulos - Senior Research Officer Publications

  • Eliza Hawkes - Clinician Scientist Publications
  • Umbreen Hafeez - PhD Student
  • Sid Menon - PhD Student
  • Fiona Scott - Program Manager Publications
  • Kerryn Westcott - Scientific Project Officer Publications
  • Uwe Ackermann - Senior Radiochemist, Department of Medical Imaging & Therapy, Austin Health (Honorary)
  • Farshad Foroudi - Director Radiation Oncology, Austin Health (Honorary) Publications
  • Sze-Ting Lee, Nuclear Medicine Physician, Department of Medical Imaging & Therapy, Austin Health (Honorary) Publications
  • Prof Michael McKay, Senior Researcher (Honorary) Publications


Cancer Single Cell Genomics Laboratory

Our research focus

Our goal is to understand cancer at a single cell level to advance cancer biomarker discovery and translation

Our laboratory specialises in using innovative single cell techniques to deconstruct tissue samples and reveal the genetic architecture of individual cells. This allows us to study heterogeneity in a range of cancers, including breast cancer, and address key biological questions:

  • How intra-tumoural heterogeneity drives tumour progression and metastasis in aggressive cancers?
  • What role do the tumour microenvironment play during metastasis?
  • How can we effectively monitor metastatic disease progression and prevent cancer recurrence?
  • Which premalignant molecular alterations are involved in breast tumorigenesis and can rare aberrant cell populations be identified for target biomarker discovery?
  • How single cell transcriptomics can be utilised to improve personalised treatment for cancer patients?

Fast facts

Metastasis refers to a disease stage when cancer cells break away and spread beyond the primary tumour to distant sites, with variable locations and volumes of organ involvement.

Created by a dynamic micro-community of cancer cells and surrounding blood, immune and stromal cells. Interactions between resident cell types can influence tumour progression and patient response to cancer treatment.

Our team

Meet our researchers

  • Dr Bhupinder Pal – Head, Cancer Single Cell Genomics Laboratory Publications
  • Paula Fuge-Larsen - Translational Research Project Officer
  • Shalini Guleria - PhD Student
  • Jordan Wilcox - Research Assistant


Tumour Progression and Heterogeneity Laboratory

Our research focus

Isolation and characterisation of circulating tumour cells

Liquid biopsies, which capture circulating tumour cells in the blood, are a useful, non-invasive way of monitoring tumour spread and drug response. Our laboratory studies the diversity and biological properties of cancer cells captured in blood. This helps improve the diagnosis of patients, predict drug response and, in the longer term, develop cancer treatments personalised to a patient’s specific cancer.  

Follow tumour progression using cellular tracking

Each cell collected from a patient’s tumour can be labelled with tags or ‘barcodes’, allowing us to determine which subpopulations of cells in the tumour contribute to metastasis, organ specificity and drug-resistance. We are particularly interested in the effect of different microenvironments or ‘niches’ on the survival of cancer cells and the progression of disease.

Test new drugs in advanced models of metastatic breast cancer

Our laboratory is interested in developing ways to test the effect of various drugs on the survival of circulating tumour cells or metastasis. In particular, we focus on testing the effect of new targeted therapies on metastatic progression.

Fast facts

Some cancer cells have the ability to spread in the body. They can invade locally to nearby lymph nodes, to the vasculature and distant organs. This process is called metastasis. The mechanisms by which cells are able to adapt to different microenvironment are still unknown, but it appears that only a few cells from a tumour will successfully grow in distant organs and cause symptoms.

Different tumour cells in a tumour can show distinct phenotypic profiles such as gene expression, proliferation, and metastatic potential.

Drugs which specifically block the proliferation, survival or invasiveness of cancer cells, by targeting specific cellular pathways.

Recent publications

Cancer Cell

BH3-Mimetic Drugs: Blazing the Trail for New Cancer Medicines.

DOI: 10.1016/j.ccell.2018.11.004

View abstract
Nature Communications

Barcoding reveals complex clonal behavior in patient-derived xenografts of metastatic triple negative breast cancer.

DOI: 10.1038/s41467-019-08595-2

View abstract
Science Translational Medicine

Synergistic action of the MCL-1 inhibitor S63845 with current therapies in preclinical models of triple-negative and HER2-amplified breast cancer.

DOI: 10.1126/scitranslmed.aam7049

View abstract

Our team

Meet our researchers