Development of multiplexing strategies for electron and super-resolution optical microscopy/

[摘要] The aim of this work is to increase the multiplexing capabilities of electron and super resolution optical microscopy. This will be done through the development of molecular-scale barcodes that can be resolved in one of the two high resolution imaging modes. In the optical domain, the number of colors available in stochastic optical reconstruction microscopy (STORM) will be increased by taking advantage of not only the spectral differences between STORM fluorophores but their kinetic properties as well. In the electron microscopy domain, the recently developed electron contrast-generating protein miniSOG will be concatenated to produce fully genetically encoded barcodes that can be resolved using standard transmission electron microscopy techniques. At the time of writing, the hardware for a STORM microscope has been assembled. Single molecule fluorescence blinking has been observed, though the imaging buffer still needs to be optimized for imaging. Concatamers of miniSOG have been generated and can be expressed in HEK cells and photo-oxidized.

[发布机构] Massachusetts Institute of Technology