Development of correlative atomic force microscopy and fluorescence microscopy to investigate cytoskeletal properties of pancreatic cancerous cell

Session

Session 5 - Super-resolution and Nanoscale Imaging

Authors

Samhitha Shivananda (1, 1), Thomas Waring (1), Marie Held (1), Lorna Young (1)

Affiliations

1. University of Liverpool

Keywords

atomic force microscopy

correlative microscopy

pancreatic cancer

cytoskeleton

structured illumination micrscopy

Abstract text

A fundamental component of cell structure and function is the actin cytoskeleton, which plays a critical role in proper cell division, cell movement and organelle transport. Determining the nanometre detail of healthy and diseased cells is essential for understanding the mechanisms of pathogenesis. Atomic force microscopy (AFM) is a powerful tool that allows for the measurement of structure topography and mechanical properties at the nanometre scale. However, it lacks specificity in structure identification, which can make it difficult to identify structures of interest within a wider context. Structured illumination microscopy (SIM) is a super-resolution light microscopy technique that allows sub-diffraction limit imaging of fluorescently probes. This allows specific identification of proteins, and importantly, structures of interest.

 

In this work, we develop a novel and innovative microscopy technique and analysis pipeline, which correlates the structural specificity of SIM and the nanometre-scale mechanical and topographical data of AFM. This approach provides us the ability to use SIM and light microscopy to identify structures of interest, and then measure the topography and mechanical data within those structures. Using this combined technique, we measure the three-dimensional details of cytoskeletal structures within pancreatic cancer cells as a disease model. The work will provide a pipeline for future users of correlative AFM and super-resolution microscopy techniques to examine many cellular sub-structures, which will provide critical insight into the functioning of cells.