Abstract
Spider silk fibres share unprecedented structural and mechanical properties which span from the macroscale to nanoscale and beyond. This is possible due to the molecular features of modular proteins termed spidroins. Thus, the investigation of the organizational scaffolds observed for spidroins in spider silk fibres is of paramount importance for reverse bioengineering. This dataset consists in describing a rational screening procedure to identify the nanoscale features of spider silk fibres. Using atomic force microscopy operated in multiple acquisition modes, we evaluated silk fibres from nine spider species. Here we present the complete results of the analyses and decrypted a number of novel features that could even rank the silk fibres according to desired mechanostructural features. This dataset will allow other researchers to select the most appropriate models for synthetic biology and also lead to better understanding of spider silk fibres extraordinary performance that is comparable to the best manmade materials.