ANSTO's human health research facilities

 

Our facilities are closely linked with our unique capabilities in:

 

 


Bioanalytics    

 

The bioanalytics facilities enable research to be conducted in the areas of cell biology, molecular biology, membrane biophysics and electrophysiology.

 

Cell biology facilities

 

The bioanalytics cell biology facilities provide critical support to all aspects of our work. Our facilities provide researchers with the necessary equipment to perform a large variety of traditional cell biological analyses as well as non-standard, experimental and technically novel applications. Making use of the established waste management procedures at ANSTO, this facility is suitable for the analysis of radioactive materials which can include contaminated materials, isotopic detection methods and radiotracers.

 

Detection methods available to researchers at the bioanalytics cell biology facility include:

 

  • Live-cell imaging
  • UV/Vis spectrophotometry
  • Chemiluminescence
  • Bioluminescence
  • Calcium imaging
  • Photo stimulated luminescence
  • Fluorescence
  • Fluorescence polarisation
  • Time resolved fluorescence
  • Gamma counting
  • Liquid scintillation analysis


The cell biology facilities make use of a suite of standard cell analysis techniques/equipment including:

 

  • Fluorescence and light microscopy including phase contrast and darkfield
  • FLA7000 high resolution multimodal scanner
  • LAS4000 cooled CCD gel documenter with diode excitation
  • Wizard 2480 ɣ counter
  • Tricarb LSA β counter
  • Spectramax M5 multimodal 12-384 plate reader
  • Flow cytometry
  • Full tissue culture facilities including long term cryo storage
  • Separate general biology facilities for tissue preparation

 

Molecular biology facilities

 

The molecular biology facilities are primarily used to quantify the expression of genes by measuring RNA and protein levels. Cultured cell lines can also be modified to over express, knock-down or knock-out genes of interest.

 

  • CFX384 real time PCR detection system for quantifying nucleic acids
  • High resolution melt analysis to detect differences in double-stranded DNA
  • T100 thermal cycler for amplifying DNA fragments
  • Gel electrophoresis equipment for separation and analysis of nucleic acids and proteins
  • TransBlot turbo transfer system for high efficiency blot transfer of proteins
  • Standard molecular biology equipment including water baths, incubators, incubator shakers, centrifuges and a Nanodrop spectrophotometer are used to create expression plasmids
  • Nucleofection for the delivery of DNA, RNA and biomolecules into difficult to transfect cell lines

 

Membrane biophysics and electrophysiology facilities

 

Equipment available to the bioanalytics group enables the study of biological membranes. One area of focus has been examining membrane proteins and their interaction with artificial lipid bilayer membranes. Electrophysiology equipment is also available to study the electrical properties of vesicles, organelles, cells and tissue sections.

 

  • Electrical impedance spectroscopy to characterise the sub-structure of lipid bilayers and cells
  • Quartz crystal microbalance with dissipation monitoring to characterise the formation of lipid bilayers
  • Conventional patch clamp setup (including Axon digitizer and amplifier)  to record single channel and whole cell currents of vesicles, cells, and tissue sections (i.e. brain and spinal cord slices)
  • Nanion automated patch clamp to record currents of vesicles, organelles and cells in suspension
  • Nanion vesicle preparation device to prepare vesicle for patch clamp and impedance spectrometry
  • Olympus BX61 water immersion microscope for imaging and monitoring of procedures during conventional patch clamp
  • CoolSNAP digital camera to record images including live-cell and calcium imaging

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Investigations with Imaging  

 

Imaging is about revealing processes and events in living matter: showing the metabolic pathways a molecule travels in cellular structures or in animals, discovering the cellular events of a disease, or finding and identifying a missing protein. Using radio-labelled molecules, imaging with Positron emission tomography (PET) or Single photon emission computed tomography (SPECT) can track events and target proteins in a cell, on a cell and outside a cell.
 

 In vivo laboratories
 Gamma Counters
The Wallac Wizard 2480 is used to measure the radioactivity in research samples such as tissues from biodistribution or samples from bioanalytical assays.  Gamma counters are located at both the ANSTO and BMRI campus.   ALS facilities Wallac Gammer Counter 420x272
 Small Animal Positron Emission Tomography (PET)

 The Siemens Inveon PET/CT is a high resolution small animal imaging system that can perform multimodal imaging resulting in fused PET/CT images. It has an axial PET Field of View (FOV) of 12.7cm and spatial resolution of 1.4mm. We are currently investigating the imaging of two mice at the same time in the system. 

 ALS facilities PET group 360x241
 Computed Tomography (CT)

CT scans used for PET/CT animal studies can be achieved in less than 15minutes and cardiac or respiratory gating can be applied to scan acquisition.

ANSTO has two of these systems, one located at our Lucas Heights campus and another located at the BMRI under the NIF node which can be used in conjunction with C-11 radiotracers produced from the Camperdown Cyclotron.

 ALS facilities PET Arvind 420x263
 Animal anaesthesia and monitoring systems
 We use inhalant isoflurane in oxygen anaesthesia throughout animal imaging sessions as well as during surgical procedures. We have available two different animal monitoring systems to record temperature, heart rate, oxygen saturation, and respiration rate; the M2M BioVet system and the Starr Life Sciences Mouse Ox.
 
 Small Animal Positron Emission Tomography /Computed Tomography (PET/CT) and Single Photon Emission Computed Tomography/CT (SPECT/CT)

The Siemens Inveon Trimodality PET/SPECT/CT is a high resolution small animal imaging system that can perform multimodal imaging resulting in fused PET/CT and/or SPECT/CT images. The PET component has an axial Field of View (FOV) of 12.7cm and spatial resolution of 1.4mm.

The SPECT component comprises 2 gamma detector heads and comes with several collimator sets: 3 dedicated to mouse imaging (1 single and 2 multi-pinholes), 3 dedicated to rat imaging (1 single and 2 multi-pinholes) and a Low-Energy, All-Purpose (LEAP) parallel-hole collimator. The collimators present a transaxial FOV varying from 90 to 110 mm for rat compared to 26 to 60 mm for mouse (ROR dependent) and axial FOVs varying from 90 to 130 mm for rat and 30 to 90 mm for mouse. The axial FOV can also be extended up to 250 mm using continuous bed motion. The measured spatial resolution ranged from 0.57 mm to 1.98 mm while the system sensitivity was found to range between 29.3 and 403.6 cps/MBq.

CT scans used for co-registration with PET or SPECT animal acquisitions can typically be achieved in less than 15minutes. This system is located at the BMRI.
 

 Stereotaxic Surgery Equipment
For the precision injection of drugs into brain regions,ANSTO has the facilities to perform stereotaxic surgery. Consisting of the stereotaxic frame, drill equipment and syringe pump the system is used in conjunction with anaesthesia and animal monitoring equipment.
 
 Femoral Artery Cannulation Equipment
Direct arterial blood sampling is necessary during complex drug multi-injection protocols and this requires cannulation of the femoral artery. ANSTO's human research infrastructure has the equipment to perform this microsurgery including microscopy, microsurgery tools, anaesthesia and animal monitoring equipment. 
 
 In vitro laboratories
 Liquid Scintillation Counters 
The Perkin-Elmer Tri-Carb 2800TR Liquid Scintillation Analyser is used for measuring radioactivity (beta particles) within binding samples or for autoradiography standards. We have one system located at the Lucas Heights campus and another located within the BMRI facilities (Tri-carb 2100TR) 
 
 Cryosectioning (micro) 
 The Leica CM3050S Cryomicrotome with disposable cutting blades is capable of creating sections as thin as 4µm thick from frozen animal tissues.  Disposable blades reduce cross-contamination that can occur when consecutively cutting different radioactive samples while the motorised sectioning capability reduces repetitive motion while maintaining reproducible section thickness and quality.  ALS facilities cyromicrotome 410x272
 Paraffin Embedding and Microtome Sectioning 
ANSTO can preserve, embed and section tissues for histological or immunohistochemical procedures. Our paraffin wax embedding system produces preserved and embedded samples in three days which can be followed by microtome sectioning to as thin as 4µm if required.  ALS facilities Wax embedding system 410x272
 Autoradiography equipment (Densitometry using phosphor imager and film) 
We operate GE Typhoon FLA7000, FujiFilm BAS-5000 and FujiFilm BAS-2500 phosphorimaging systems routinely for detecting 125I,14C, 18F, 123I, 99mTc and 111In using Fujifilm phosphorimaging plates.  Additionally for lower energy isotopes (e.g. 3H) we have film based methods using a BioRad Gs-800 Calibrated densitometer. This can be used to detect/quantify radioactive sources with spatial resolution within in vitro preparations. 
 
 Dark room 
Dark rooms have manual and automated film developing infrastructure. Films are archived using a BioRad Gs-800 Calibrated densitometer.
 
Cell Harvesters for Binding s tudies 
Brandell 24-Sample, Semi-Auto Cell Harvesters allow rapid washing and throughput of cell binding assays.    ALS facilities cell harvester

 

                                                                  

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Radiochemistry    
 

The radiochemistry capabilities and facilities are primarily devoted to developing novel radioligands for imaging (PET, SPECT) and targeted radioisotope therapy.  Other areas of interest are developing technologies to incorporate isotopes through molecular design, use of conjugates and novel macrocycles. The radiochemistry team utilise a diverse suite of radiochemistry and synthetic chemistry facilities and capabilities to deliver world class research.

 Radiochemistry Laboratories (more than 15)
Located both at Lucas Heights and Camperdown
17 remote manipulation radioisotope handling facilities (Hot Cells)
16 Lead lined fume hoods 
Radioisotope access 
Dedicated research cyclotron (18 MeV) at the Camperdown facility for PET radioisotopes (18F,11C)
64Cu, 86Y, 89Zr and 124I through the cyclotron partnership
Two in-house 68Ge/68Ga generators
Commercial sources (123I,125I, 131I, 67Ga, 68Ge, 177Lu)
Custom irradiations in the OPAL Reactor for SPECT and therapeutic radioisotopes.
Canberra HPGe Gamma Spectrometer for radionuclidic purity determinations 
Automation modules and new radiolabelling technology platforms 
GE Tracerlabs FxFn (x3)
Nuclear Interface FDG module
Synthra RN Plus (x4)
Synthra MeI Plus (x2)
Iphase Flexlab (x3)
In house built automation modules
Microfluidic chemistry is investigated using an Advion NanoTek LF
Microwave chemistry is investigated using a CEM Discover 
Radiolabelling of peptides and proteins 

Dedicated biomolecule labelling laboratory
Gradient HPLC: ion exchange, affinity, size exclusion, (Waters) 

  • Dedicated QC and separation systems
  • Assorted interchangeable detectors: Variable single and dual wavelength, UV-Vis PDA, Evaporative light scattering, conductivity and refractive index
  • Scintillation high activity and high sensitivity radio-flow detectors
Dialysis and ultrafiltration equipment
Freeze drier (FTS, FlexiDry)
Minus 80 freezer (Thermo)
Heating and cooling centrifuge (Sigma, 2-16KC)
Heated centrifugal concentrator (Eppendorf Concentrator Plus)
Ion-Chamber (Capintec, CRC) 
Radiometabolite analysis 
Ultrasonicator (Misonix S4000) and centrifuge for sample preparation
Gamma counter (Wallac Wizard 2480) for quantification
Radio TLC analysis using a phosphorimager (Fuji Bas5000) or RadioTLC scanner (Berthold Tracemaster 20)
Radio HPLC using UV visible, Posi-RAM (Model 3 IN/US systems) and Gamma-RAM (Model 4 IN/US systems) detectors
Shimadzu-ABSciex triple quadruple LC-MS-MS system dedicated for radioactive samples 
Radioligand analysis 
Dedicated QC HPLC systems (Reverse phase, normal phase, ion exchange, affinity, size exclusion)

Variable wavelength detectors (Dual and PDA)

Scintillation high activity and high sensitivity radio-flow detectors
 

Radio TLC/iTLC using a phosphorimager, radio TLC scanner or Wallac 1470 gamma counter. 
Synthetic Chemistry Laboratories 
3 dedicated wet chemical laboratories
14 fune hoods 
Synthetic Chemistry Characterisation and Analysis 
Bruker AV400 NMR spectrometer with BACS60 Autosampler
LC-MS (Waters Micromass ZQ) for routine analysis and reaction monitoring
Automated Flash Chromatography System (Grace Reveleris)
4 HPLC Systems

- Dedicated QC System (Waters Alliance 2695)

- QC and Prep HPLC Systems (Waters)
- Dedicated SEC / ion-exchange semi-prep HPLC System (Shimadzu)
 

Agilent Cary 100 UV-Vis Spectrometer with Peltier Multi-cell
Bruker Alpha Diamond ATR IR spectrometer
AA-10R Automatic Polarimeter
Eppendorf Centrifuge 5430
5 Bay Solvent Purification System (Braun)
KEM AT-610 Autotitrator
7 Buchi Rotary Evaporators
Christ Alpha 2-4 LD Plus Freeze Dryer
CEM Discover microwave reactor 

 

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Radiation Technologies   
 

The Radiation Technology  group operates cobalt-60 irradiators for small scale irradiation studies to a wide range of doses and dose rates. Our unique capabilities enable us to accurately deliver high doses of radiation with a precision not achievable in industrial-scale irradiators. Controlled dose rates and temperatures allow users to test ideas, develop applications and perform irradiations under non-standard conditions.

Gamma Technology Research Irradiator (GATRI) 
60Co wet storage batch irradiator
Varying dose rates from 0.1 to 4 kGray.h-1
Delivery of dose from 10 Gray to 106 Gray
Temperature irradiations from –78°C (under dry ice) to ambient
Wet chemistry lab with Bruker Electron Spin Resonance spectrometer and laminar flow cabinet
Sample / product size to 1m3 
Gamma Irradiation Suite (GIS) 
Irradiation and biology rooms for radiobiology research
60Co collimated beam (Theratron), mGy to Gy
Beam attenuators
PC1 biology lab 
Gammacell 220 (GC220) 
60Co dry storage self-shielded irradiator
Fixed dose rate of 2 kGray.h-1
Delivery of dose from 10 Gray to 106 Gray
Temperature irradiations from –78°C (under dry ice) to ambient
Sample chamber dia. 150mm h. 200mm
Access for thermocouples
Sample / product size to 2 litres 

 

 

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