infocus #66 June 2022 3D Nanoanalysis of the Tumour Microenvironment: Subcellular Ultrastructure of Model Systems

Many cancer tumours have a hypoxic core, I wanted to determine ultrastructural differences of the tumour regions using a 3D cell culture model. When cells lack oxygen, they metabolise differently via hypoxia inducible factors (HIF). I therefore compared non hypoxic outer regions and hypoxic core regions of the spheroid culture model of HIF-1 or HIF- 2 knockouts using resin embedded sections and transmission electron microscopy.

The aim of my project was to look at the subcellular structural changes in different hypoxia inducible factor (HIF) knockouts.

I used resin-embedded spheroid knockouts that were stained with osmium and uranyl acetate, they were grown by Luke Thornton and sectioned by Denise McLean. Using these spheroids, I used a transmission electron microscope (TEM; Tecnai-12), to image cross-sections through the core of the spheroids.

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