Towards more dose efficient cryogenic electron microscopy of biological samples

Cryo-EM has been a powerful tool in unraveling the protein structures and their functions, providing unprecedented insights into the building blocks of life. In this thesis, studies have been carried out to develop new cryo-EM methodologies, which enable scientists to overcome existing challenges and allow for higher dose efficiency imaging and diffraction techniques. It presents different studies that incorporate novel approaches to improve the cryo-EM workflow, ranging from the introduction of a standard protein, investigation of the size limitation in single particle analysis, offering a solution to overcome charging of vitreous samples, developing advanced detectors, to describing alternative schemes to improve the dose efficiency. Cryo-EM has revolutionised structural biology over the last decade. Allowing for the visualisation of smaller biomolecules from fewer numbers of particles such that their dynamic assemblies could be better observed and understood, could revolutionise structural biology yet again in the decade to come. This study might contribute towards this.