Imaging
Seeing is believing. We offer access to advanced microscopy equipment and consultation for Sydney-based users.
About
CMRI’s Imaging Facility aims to enable world-leading research by providing comprehensive support for light microscopy techniques. The centre supports experimentation from conceptual planning to data analysis and publication through training, guidance, and instrument access.
Reach out to Will Hughes for details, training, consultation and booking information.
Instruments
We have a wide range of microscopes, capable of multi-modal imaging of biological samples ranging from organs (whole murine brain), tissues to organoids, cell monolayers, single cells and single molecules. We have brightfield, fluorescence, live cell imaging, laser scanning confocal, spinning disc confocal, light-sheet, fluorescence lifetime imaging, semi-super and super resolution and slide-scanning instruments.
Techniques
We offer expertise in a wide range of imaging techniques such as histology, direct and indirect fluorescence microscopy, live cell brightfield, widefield fluorescence and confocal imaging including FRAP, FLIM & FLIM-FRET. In addition, we can assist with cytogenetic imaging with Metasystems Metafer and volumetric imaging with lightsheet, confocal or super-resolution Tau-STED imaging.
Analysis
We offer expertise in image analysis particularly with custom creating scripts for batch image analysis. We have eight image analysis workstations with Oxford Instruments Imaris, MetaSystems Metafer and ISIS, SVI Huygens, Zeiss Zen, Leica LASX, ImageJ/FIJI, Cell Profiler, Ilastik, and QuPath.
Examples
Fluorescence Microscopy
Fluorescence microscopy is a workhorse technique for localizing cellular components. Our Zeiss AxioImager Microscopes give researchers advanced widefield fluorescence capabilities including Z-stacks, tiling, and ApoTome imaging. Our Zeiss AxioImager microscopes with MetaSystems Metafer software enable researchers to automate imaging and analysis of metaphase chromosomes and interphase nuclei.
Live Cell Imaging
Viewing cells in their living state provides a wealth of information not available from fixed specimens. Live cell imaging involves the collection of images at successive time intervals under conditions that support cell culture. Four of our microscopes support live cell imaging: a Zeiss Cell Observer for transmitted light and widefield fluorescence imaging, a Zeiss Cell Observer Spinning Disc confocal and Zeiss LSM 880 Airyscan for confocal and super-resolution imaging and a Leica Stellaris 8 FaLCon STED for confocal, Lifetime and super resolution imaging of live cells.
Confocal Microscopy
Confocal microscopy enables researchers to visualize cellular structures in three dimensions. Our laser-scanning confocal microscope, Zeiss LSM 880 Airyscan, has seven laser lines, selectable emission range and spectral unmixing capabilities. The Zeiss Cell Observer SD microscope supports multichannel confocal imaging of live cells using spinning disk technology. Our Leica Stellaris 8 FaLCon STED has a supercontinuum white light laser providing pulsed, tuneable excitation energy from 440-790nm and 5 hybrid (3x HyD S, 1x HyD X, 1x Hyd R) detectors spread across 400-800nm.
Super-Resolution Microscopy
Visualising separate objects smaller than the theoretical limit of optical resolution is called super-resolution.
With the Zeiss LSM 880 Airyscan, standard fluorescent samples and live cells can be imaged with as low as 120 nm resolution. With the Leica Stellaris 8 FaLCon STED using a 775nm depletion laser and lifetime-gating (Tau-STED) fluorescent samples can be imaged with as low as 30nm (x&y) by 100nm (z) resolution.
Lightsheet Imaging and Stereomicroscopy
Lightsheet imaging is an optical microscopy technique in which a section of a sample is illuminated with a thin sheet of light, resulting in robust, fast and gentle three-dimensional imaging of large specimens. When combined with a tissue clearing sample preparation technique, large tissue samples or whole-mount embryos up to 5 mm thick can be imaged using the Zeiss Lightsheet Z.1/7 microscope. Optically sectioning fluorescent samples using Lightsheet microscopy can replace traditional serial sectioning in a fraction of the time while retaining the overall three-dimensional structure of the sample. Using a more traditional microscopy technique, the Zeiss SteREO Lumar.V12 Fluorescence Stereomicroscope enables imaging large samples such as embryos, zebrafish and whole tissues.
Have a project in mind?
We’d love to work with you.
Our Instruments
Zeiss Lightsheet Z.1/7 Microscope
The Zeiss Lightsheet Z.1/7 is a fluorescent microscope designed to image larger specimens ranging in size from 0.5 mm to 5 mm. Specimens are able to be rotated 360 degrees, allowing the entire specimen to be imaged from different views before being digitally reconstructed. We have 5x and 20 objectives and specialized sample chambers are available for live samples or samples that have been cleared using certain tissue clearing techniques, including SCALE. The Lightsheet Z.1/7 is accompanied by a specialised image analysis workstation.
Zeiss SteREO Lumar.V12 Fluorescence Stereomicroscope
The Zeiss SteREO Lumar.V12 stereomicroscope is capable of high-resolution three-dimensional imaging over a large field of view. This makes it the ideal system for imaging larger samples such as embryos, zebrafish and whole tissues. It has brightfield reflected and transmitted light capabilities, as well as a range of filters (DAPI, GFP, YFP, RFP) allowing fluorescence imaging. The Zeiss SteREO Lumar.V12 stereomicroscope is a fully motorised with a magnification range from 9.6x to 120x and is capable of multichannel acquisition, Z-stacks, time lapse and tiling experiments.
Zeiss LSM 880 Airyscan Laser Scanning Confocal Microscope
Laser scanning confocal microscopy is an essential method in cell biology research for its ability to optically section and reconstruct cellular samples in three dimensions. The Zeiss LSM 880 is equipped with the Airyscan detector. This microscope allows visualization of intracellular structures in higher resolution, up to the super-resolution level.
Zeiss Cell Observer SD Spinning Disk Confocal Microscope
The Zeiss Cell Observer SD microscope uses Yokagawa spinning disc technology to generate 3D confocal images extremely quickly..
Zeiss Axio Observer Widefield Live Cell Microscope
Visualizing living cells offers a dimension to experiments that fixed (preserved) specimens can’t provide. Live cells can be imaged over the course of multiple hours or days. Cells can be imaged unlabelled or tagged with fluorescent proteins.
Zeiss Axio Imager Z2 with ApoTome.2 Widefield Upright Microscopes
Our two Axio Imager upright fluorescence microscopes are the ‘workhorses’ of the facility, allowing researchers to capture images of a wide range of fluorescently-labelled cell samples. The ApoTome.2 instrument allows the reconstruction of images in three dimensions with enhanced resolution and contrast.
MetaSystems Metafer Chromosomal Analysis Microscopes
CMRI researchers pioneered the use of automated metaphase chromosome imaging in telomere research using the Metafer automated fluorescence microscopy system. Our two Metafer systems accelerate research by automating fluorescence microscopy of cancer and normal cells, enabling imaging in a few hours cell that would otherwise have taken weeks to complete.
Leica Stellaris 8 FaLCon STED
Our Leica Stellaris 8 FaLCon STED is a fully integrated system enabling confocal, fluorescent lifetime and super-resolution imaging in fixed or live samples.
It has a 405nm and supercontinuum white light laser 440-790nm and 5 hybrid (3x HyD S, 1x HyD X, 1x Hyd R) detectors spread across 400-800nm.
Fluorescent lifetime imaging by FaLCon, Fast Lifetime Contrast is a fully integrated form of time correlated single photon counting using the WLL and HyD detectors.
Super Resolution Imaging is possible in fixed and live samples using Tau-STED, lifetime-gated stimulated emission depletion microscopy using a 775nm depletion laser. The spatial resolution of Tau-STED is about <30 nm for fixed specimen and <70 nm for live specimens compared to 200 nm in a regular confocal microscopy.
