Using Live Cell STED Imaging to Visualize Mitochondrial Inner Membrane Ultrastructure in Neuronal Cell Models

Our research program focuses on structural and functional aspects of mitochondria, particularly as they relate to aging related dysfunction. To this end, we use a broad array of biophysical, biochemical, and structural techniques to explore the basic mechanisms of mitochondrial aging and to develop therapeutic interventions to preserve and restore mitochondrial health. Mitochondrial function and structure are inextricably related.

Standard light microscopy techniques are suitable for measuring large scale mitochondrial features like basic morphology and network structures. Analyzing mitochondrial ultra structure or features that require higher resolution than afforded by traditional optical microscopy is typically done using electron microscopy or tomography on fixed samples. Super resolution microscopy is a fluorescence based imaging technique that measures features beyond the diffraction limit.

For mitochondria, this includes measurements of nano scale protein distribution and cristae architecture. The stead imaging protocol described here allows researchers to measure the detailed structure of the inner membrane in living cells. Live cell imaging allows us to observe and quantitatively analyze complex features of cristae under physiologically relevant conditions without the need for sample fixation.

This will give researchers novel insights into the dynamic changes that occur in ultra structural features and their temporal responses to stressors and pharmacological compounds.