Live Cell and Dynamic Imaging

WEHI Centre for Dynamic Imaging provides precision imaging across a broad scale, from single cells and tissue slices to whole organisms using both fluorescence and bioluminescence modalities.

First published: 15/12/2014

Updated at: 16/12/2014 at 3:02 pm

When it comes to getting the best images from microscopes, most VPTN facilities will push their instrumentation to the limits to improve precision.

The Centre for Dynamic Imaging at the Walter and Eliza Hall Institute is one that goes to great lengths to provide precision imaging across a broad scale, from single cells and tissue slices to whole organisms using both fluorescence and bioluminescence modalities.

Precision Imaging including Live Cell

Among the variety of equipment available in this PC2 imaging platform are state-of-the-art widefield and confocal microscopes for 2, 3 and 4-dimensional imaging as well as the IVIS Spectrum, which is a preclinical non-invasive imaging system. Images are usually acquired over 1-10 minutes with five field of view options ranging from 4 cm to 25 cm bioluminescence and fluorescence in living animals. The equipment also allows consecutive images to be captured in a slow kinetic readout to track substrate uptake in tumours over time, but is most commonly used for monitoring cellular and genetic expression patterns in longitudinal studies.

For research into cancer, chronic inflammatory disease and infectious disease

Lab manager Dr Kelly Rogers welcomes researchers from other non-profit organisations as well as Walter and Eliza Hall Institute researchers, to use the equipment once they’ve been trained.

“The Institute’s Dynamic Imaging laboratory is dedicated to research into cancer, chronic inflammatory disease and infectious disease. It offers researchers access to state-of-the-art equipment and training in advanced optical imaging techniques so they can work independently or within a collaborative environment, which enables the exchange of know-how and experience in the application and analysis of imaging experiments,” she said.

“The laboratory has staff with expertise covering all aspects of multidimensional data acquisition and analysis, including three-dimensional reconstruction of cell and tissue structure and in vivo preclinical fluorescence and bioluminescence imaging.

“When researchers explain the outcome they’re seeking, we can advise them how to get the best images and then assist them with the quantitative analysis of their data,” she said.

Dr Rogers own expertise is in method development for calcium imaging, live cell and confocal microscopy and preclinical imaging. Dr Whitehead, who is a physicist and computational scientist by background, is an expert in image processing and analysis, and is helping to drive the application and development of bioimage informatics within the lab.

Her colleague, Mark Scott has a histopathology background with expertise in electron microscopy and intra-vital imaging. He also has good experience with a number of whole-mount imaging techniques such as OPT as well as a variety of software analysis platforms.

Two-photon imaging

Mark’s passion is in ‘two-photon imaging’, which uses a pulsed infra-red laser to excite molecules which means we can image through thicker tissue samples, giving us more structural information.

“Through specialised techniques we can also image collagen and elastin fibres without the need to stain them, to look at specialised tissue damage or to look at the effect of therapeutic drug therapy”.

“This is particularly exciting for heart transplant imaging. Sometimes heart valves are grown from scratch using stem cells onto a collagen matrix structural scaffolding. The two-photon allows us to image the collagen without having to stain to find out where a fluorescent stem cell is moving to,” he said.

Current imaging equipment at WEHI Centre for Dynamic Imaging includes:

Confocals

  • Zeiss LSM 780 with a 32-channel GaAsP array detector.
  • Leica SP8 confocal with 8 kHz resonant scanner and the high sensitivity HyD detectors.
  • Olympus FV1000 confocal and multiphoton system with dual head.
  • Zeiss LSM 5 Live Line Scanning Confocal

Other microscopes

  • Several widefield fluorescence microscopes for live cell imaging, including a Zeiss AxioObserver, Nikon TiE and Deltavision Elite Microscope.
  • FLIM microscope based on the frequency domain.
  • An Olympus SZX16 fluorescence stereomicroscope
  • A BD Pathway 855 Bioimager for high-content screening
  • A Xenogen IVIS Spectrum
  • Several high-end 64-bit Image analysis workstations with a range of software packages, such as IMARIS, MetaMorph, ImageJ/FIJI and MatLab.

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