Geometry-preserving expansion microscopy microplates enable high-fidelity nanoscale distortion mapping
Rajpinder S. Seehra, Samantha J. Warrington, Benjamin H.K. Allouis, Thomas M. D. Sheard, Michael E. Spencer, Tayla Shakespeare, Ashley J. Cadby, Daniel Bose, David Strutt, Izzy Jayasinghe
Abstract
Expansion microscopy (ExM) is a versatile super-resolution microscopy pipeline, leveraging nanoscale biomolecular crosslinking and osmotically driven swelling of hydrogels. Currently, ExM is a laborious and skill-intensive technique, involving manual handling of the hydrogels that can compromise the integrity of the gels and capacity to track gel isotropy, hence diminishing reproducibility. We have developed a 3D-printable microplate system to contain the entire ExM workflow within each well, enabling in situ image acquisition and eliminating the need for direct handling of the hydrogels. The preservation of the gel geometry and orientation of the microplate wells enables convenient tracking of gel expansion, pre- and post-ExM image acquisition, and distortion mapping of every cell or region of interest. We demonstrate the utility of this approach with both single-color and multiplexed ExM of cultured HeLa cells and dissected pupal Drosophila melanogaster wing tissue to reveal distortion-prone structures ranging from sub-cellular organelles to micron-scale tissue regions.