NWO ECHO Project Jacob Hoogenboom approved

Title: "Optimized electron-molecule interactions for near-molecular resolution light and electron microscopy". The researchers propose a novel approach: fluorescence microscopy using a beam of energetic electrons. This will allow measuring molecular positions in the structural landscape at electron microscopy resolution. Their approach is enabled by two unconventional steps: (i) Fluorescent molecules will be excited in an electron microscope using low-energy (1-50eV) electrons, probing resonant and near-resonant intramolecular excitation regimes. (ii) Encouraged by recent initial observations of electron-excited fluorescence from green fluorescent protein (GFP) under vacuum, will optimize fluorescent proteins for fluorescence microscopy with focused electron beams. Thus, we will enable electron-excited fluorescence from organic fluorescent molecules commonly used as bio-molecular labels for immuno-targeting, as well as from optimized fluorescent proteins.

The outcome of molecular reactions is intricately determined by the local structural environment on which the reaction takes place. With superresolution fluorescence microscopy, awarded the 2014 Nobel Prize, molecules can be localized with 20nm resolution. However, the underlying structural context remains invisible, while this local structural detail is crucial in understanding how biological molecules function, e.g., within cells, and how they dysfunction during diseases. With electron microscopy, structural detail can be imaged at nanometer resolution, but the molecules remain invisible. To localize interacting molecules with very high precision within their sub-cellular context is a key challenge in biochemistry.

Applicants: Jacob Hoogenboom (TU Delft), Ben Giepmans (UMC Groningen)

ECHO grant NWO-Chemische Wetenschappen, € 257k (1 PhD in Delft)

This project is one of the five projects which were granted out of 69 applications.