Líffræðifélag Íslands - biologia.is
Líffræðiráðstefnan 2021
Erindi/veggspjald / Talk/poster E51
Höfundar / Authors: Sigríður Rut Franzdóttir (1), Zophonías Oddur Jónsson (2)
Starfsvettvangur / Affiliations: Háskóli Íslands
Kynnir / Presenter: Stefán Eggertsson
Conventional light microscopes are limited to a resolution of around 250 nm. However, many organelles and cell components cannot be adequately imaged given this limit. Super resolution methods aim to resolve this issue. For this project we aim to use two different methods of super resolution to study the subcellular localization of Pontin and Reptin in the fruit fly nervous system and sites of interaction with candidate protein partners. The methods of choice are RESOLFT microscopy, for which we are making fluorescently tagged fly lines, and Expansion Microscopy (ExM), the focus here. ExM is a recent method for super resolution and involves the embedding of a specimen in a swellable hydrogel for subsequent isotropic physical expansion, pulling proteins or nucleic acid molecules apart. This results in greater physical distance between two fluorophores and, depending on the expansion factor, pulls them into resolvable distance from each other. With this sample preparation, samples can be imaged at super resolution with conventional wide-field or confocal microscopes. Compatible with off-the-shelf antibodies, genetically encoded fluorophores and adapted to many different sample types, particularly cell culture and brain slices, this method has found wide use for researchers needing a more detailed view of their specimen. Here, I will give an overview of the variation of existing methods and of more recent advancements.