Oral Presentation 26th ACMM “2020 Visions in Microscopy”

Large scale 3D imaging of the extracellular matrix in mouse and human tissue (#147)

Howard J. Vindin 1 2 3 , Angela R.M. Kurz 4 , Brian G.G. Oliver 3 5 , Anthony S. Weiss 1 2 6 7
  1. Charles Perkins Centre, The University of Sydney, Sydney, NSW, Australia
  2. School of Life and Environmental Sciences, The University of Sydney, Sydney, NSW, Australia
  3. Woolcock Institute of Medical Research, The University of Sydney, Sydney, NSW, Australia
  4. The Centenary Institute, Camperdown, NSW, Australia
  5. School of Life Sciences, University of Technology Sydney, Sydney, NSW, Australia
  6. Bosch Institute, The University of Sydney, Sydney, NSW, Australia
  7. Sydney Nano Institute, The University of Sydney, Sydney, NSW, Australia

In recent years there has been substantial progress in terms of our understanding of the extracellular matrix (ECM) as not just a structural support but a critical regulator of cellular function and organ homeostasis. Regulation of the ECM in development and disease is a complex process involving reciprocal interactions between a variety of cells and their surrounding matrix which is poorly understood. While histological and fluorescence studies on thin tissue sections have provided us with greater insight into these processes; the preparation of these sections may introduce artefacts that are difficult to detect and by their very nature only represent a tiny snapshot of the processes occurring within the whole organ.

In order to address these issues to better visualise structural changes in the ECM that occur in development and disease we have developed a variant of the SWITCH fixation[1] protocol that provides improved structural preservation and allows for the visualisation of both collagen and elastin through second harmonic generation and endogenous fluorescence respectively. Utilisation of this method has allowed for the visualisation of whole mouse organs and embryos at sub-micron resolution giving unprecedented detail in terms of the structural organisation of cells and their surrounding ECM. We have further used this methodology to investigate structural changes occurring within the airways and alveoli of human lung in end stage disease and have been able to visualise fine branching of the smallest airways as they extend into alveolar ducts which we were able to observe at depths of up to 2mm.

  1. [1] E. Murray, J.H. Cho, D. Goodwin, T. Ku, J. Swaney, S.Y. Kim, H. Choi, Y.G. Park, J.Y. Park, A. Hubbert, M. McCue, S. Vassallo, N. Bakh, M.P. Frosch, V.J. Wedeen, H.S. Seung, K. Chung, Simple, Scalable Proteomic Imaging for High-Dimensional Profiling of Intact Systems, Cell 163(6) (2015) 1500-14.