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Microscopy and histology

Microscopy

Microscopes are used to produce magnified visual or photographic images of small objects. Samples can be easily visualised without labelling e.g. brightfield, phase contrast. Further characteristics can be determined via fluorescence labelling.

In epi-fluorescence/widefield microscopy, the whole specimen is bathed in light and all parts of the specimen are excited at the same time. Therefore, out of focus light originating from above and below the focal plane (secondary fluorescence) is captured along with light originating from it.

Confocal microscopy uses a screen with a pinhole to exclude out of focus light and produce a sharper image. This can generate an image through a point scanning process, where it scans each pixel individually and compiles a digital image. It can also be used to obtain images of planes at various depths within the sample (known as optical sectioning or z stacks) and then create a 3D image. In spinning disk confocal there are numerous pinholes which are moved by means of a rotating disk. Therefore, the entire image is collected at the same time. This increases speed, but sacrifices resolution compared to point scanning confocal microscopes.

Spectral unmixing can be used to eliminate autofluorescence and distinguish overlapping signals when multiple fluorophores are used. This process often increases sensitivity and accuracy in multi-parametric immunofluorescence.

Some epi-fluorescence and confocal microscopy applications include:

  • Immunolabelling
  • Localisation of proteins
  • Live cell imaging of cellular processes such as endocytosis, phagocytosis, protein trafficking
  • Cell-cell interactions
  • Complex techniques for measuring cell complexity (fluorescence recovery after photobleaching; FRAP), or to determine molecular proximity (fluorescent resonance energy transfer, FRET).
  • 3D reconstruction

We conduct regular training sessions on all our microscopy equipment.

For further details please contact

Connie Jackaman: connie.jackaman@curtin.edu.au

Histology

Equipment

  • Leica processing and embedding station
    • for paraffin blocks
  • Dewaxing and staining workstation
  • Microme cryostat

We conduct regular training sessions on the above histology equipment.

For further details please contact:
Connie Jackaman: connie.jackaman@curtin.edu.au

360° microscopy facility

Immerse yourself in a 360° virtual experience of our state-of-the-art Microscopy and Imaging Laboratory facility.