Applications

Potential applications

Histology

  • Special stains to identify specific cells: Tartrate-resistant Acidic Phosphatase (TRAP stain. Osteoclasts), Oil Red O (lipid), periodic acid Schiff’s stain (glycogen), Congo Red (amyloid), Masson trichrome (muscle, collagen fibres, fibrin) and Perl’s prussian blue (hemosiderin).
  • Antibody work-up: Optimise antibodies including determining the optimal fixative, antigen retrieval method, and dilution of antibody. Recent examples include IFNβ, pAKT, pStat3, IL6, and immune cell markers.
  • Supply a specialist IHC service for a range of frquently used antibodies including Ki67 (cell proliferation), cleaved caspase 3 (cell death), CD31 (endothelial cells), CD8 (T-cell lymphomas, cell surface of cytotoxic subpopulations of T-cells), BrdU (DNA synthesis), E-Cadherin (calcium dependent cell-cell adhesions, invasion/growth of epithelial cancers), PCNA (cell proliferation), pERK (cell proliferation, differentiation, motility and death).
  • OPAL staining for 8-color fluorescence imaging. We have developed a T cell panel and a pan immune panel.

Optical microscopy

  • Examine the distribution of proteins in cells of interest without out-of-focus blurring using confocal microscopy.
  • Evaluate cellular processes including cell-cell interactions, mechanisms of cell death, cell proliferation and differentiation, and intracellular signaliing pathways in real time using time-lapse imaging.
  • Investigate the mechanisms underlying asymmetric cell division and cell fate determination in real-time over multiple generations with low phototoxicity using the spinning disc confocal microscope.
  • Utilise superresolution microscopy to examine the distribution of specific proteins beyond the resolution of routine confocal microscopes.
  • Examine the mechanisms of cancer metastasis, the interaction of immune cells with cancer cells, the development of tumours over time, or the efficiency of chemotherapeutic agents in destroying cancer cells in vivo with the multiphoton microscope.
  • Use the SIM scanner on the multiphoton microscope to bleach regions of interest or photoconvert fluorescent proteins in a designated region to examine protein kinetics.
  • Perform lineage tracing using photoconvertible dyes with the confocal or multiphoton microscopes.
  • ‘Capture’ specific cells such as cancer cells without contamination from surrounding cells using laser capture microscopy. Extract the DNA or RNA from these cells for downstream processing.

Electron microscopy

  • Pinpoint the distribution of proteins of interest using immuno-gold labelling (TEM and SEM).
  • Visualise subtle ultrastructural changes due to loss of a particular protein (mouse knock-out models) using TEM. For example, quantify changes in granule number and/or morphology upon loss of granzyme A or B or perforin, or examine changes in platelet architecture upon loss of the protein under investigation.
  • Image modes of cell death (autophagy, apoptosis, necrosis) using TEM
  • Locate structures or examine cells of particular interest such as virions, immune cells within a tumour, tight junctions, stem cells, myelin sheaths, FACS sorted cells, bone marrow architecture, laminin, etc. using TEM
  • Analyse changes in the surface architecture of cells or tissues in 3D using SEM
  • Visualise surface changes due to loss or overexpression of a protein of interest, or due to treatment of cells with various experimental regimes using SEM.

Potential users

  • Research students (Honours, Masters, PhD), post-doctoral scientists, research assistants, and group leaders.
  • Commercial companies

Facility use

Use of the facility is available to all VCCC members and external users, subject to availability.

For further information about accessing the Centre for Advanced Histology and Microscopy please contact us.

Contact

A/Prof Sarah Ellis

Phone: 03 8559 7822

Email: [email protected]

Head: Centre For Advanced Histology And Microscopy (CAHM)