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Label-free 3D morphological nanoscopy for studying sub-cellular dynamics in live cancer cells with high spatio-temporal resolution

Periodic Reporting for period 3 - 3D-nanoMorph (Label-free 3D morphological nanoscopy for studying sub-cellular dynamics in live cancer cells with high spatio-temporal resolution)

Reporting period: 2022-07-01 to 2023-12-31

There is a need for label-free nanoscopes (i.e. microscopes that do not use fluorescence labels and support <250 nm resolution, which is beyond the diffraction barrier) so that living biological cells can be imaged in their most natural and unperturbed state possible. This need is being addressed here. The data from such microscopes is expected to hold vital clues to sub-cellular mechanisms of life-critical functions of cells and therefore assist in disease understanding and eventual drug design that address such mechanisms. The technology developed in this project will be used for imaging autophagy processes, which are known to be of significant interest in cancer progression and intervention.

The overall objectives are

• To kick-start a new research field at the nexus of nanoscopy and non-linear inverse scattering.
• To harness the advantages of non-linear inverse scattering into the field of label free nanoscopy, overcoming the long-standing limitations of diffraction barrier.
• To demonstrate 3D-nanoMorph’s immense utility in biology and life-sciences by imaging the degradation of sub-cellular organelles in real-time during autophagy in cancer cells, opening up new possibilities for cancer research.

3D-nanoMorph’s objectives will be achieved through devising a novel imaging system, developing novel inverse scattering algorithm, and applying the technology for imaging autophagy of live cancer cells.
Invention disclosure of certain imaging system designs have been submitted. Several articles on computational microscopy have been published. We have investigated different illumination options as well as instrumentation options. We have developed a platform design for the instrumentation. We have been investigating different algorithm options for the microscope. We have also been implementing other label-free imaging techniques for benchmarking the performance of our technique when it is eventually developed.
At the end of the project, we expect to develop a label-free nanoscopy technology, i.e. an imaging technology that does not use any form of fluorescent labels and provides a resolution superior to that supported by conventional label-free optical microscopes. We would have demonstrated the principle of symbiotic development of instrument and algorithm for achieving results beyond the state-of-the-art. We will have demonstrated the results on living cells and correlated our results with exogeneous labels in the cells that are helpful for studying autophagy.