Presynaptic Actin Nanostructures: A Reproducibility Case Study

Abstract

In an effort to assess the reproducibility of bioimage analyses in current publications, we took part in a Global BioImage Analysts Society (GloBIAS) initiative to try and reproduce results from published articles. We attempted to reproduce core findings from the work of Bingham et al., which investigates the actin organisation in presynaptic structures by using diffraction-limited and super-resolution microscopy. While the original paper unveiled clear biological insight, it lacked sufficient detail in the bioimage analysis methodological approach, limiting the depth of reproducibility we could achieve. Through frequent contacts with the corresponding author, we managed to replicate qualitative aspects of the analysis of actin nanostructures in bead-induced presynapses. We performed image reconstruction from super-resolution microscopy data, automatic image registration and visual inspection, followed by manual annotation of structures of interest in [~]35 images. Our experience with this exercise highlights the importance of transparent data sharing and accessibility, as well as the need to adhere to bioimage analysis standards that ensure the reproducibility of nowadays complex biological image analysis studies. It also shows how crucial interdisciplinary collaboration is, since many biology labs are simply unaware of these standards, which are often easy to implement and would likely be widely adopted if their value was better understood or known.