Grussmayer Lab

Department Bionanoscience

Principal investigator Kristin Grussmayer

E-mail address k.s.grussmayer@tudelft.nl

Website www.tudelft.nl/grussmayerlab

Single-molecule super-resolution imaging of Huntington’s Disease

Suitable as a BEP? Yes

Suitable as a MEP? Yes

Suitable as an Academic Research Project? Yes

Techniques:

Single-molecule super-resolution microscopy
Cell culture using cellular HD models
Microscopy in mammalian cells or in vitro single-molecule pull-down assay
Working with purified huntingtin protein and in vitro phase separation assays

Huntington’s Disease is a progressive and incurable brain disorder, caused by a single mutation in the huntingtin gene. Due to this mutation, the huntingtin protein has more than 35 glutamine repeats at its N-terminus, leading to an expanded intrinsically disordered region associated with aggregate and inclusion body formation in neurons. The mechanisms driving neurotoxicity remain however unclear, in part because early aggregate formation is challenging to study due to the diffraction limit of light microscopy. In addition, the possible role of phase separation during this aggregate formation has yet to be established. We aim to understand the dependence of aggregate formation on the protein sequence and cellular environment, with a focus on quantitative characterization of physico-chemical and mechanical properties using a combination of single-molecule fluorescence super-resolution and label-free imaging. Together, these approaches will provide new insights into the mechanisms driving early huntingtin aggregate formation and their maturation/ageing, offering potential towards novel therapeutic approaches for Huntington’s Disease.

(2024-2025) Self-labelling Protein Tags for Quantitative Super-resolution Live-cell Imaging

Supervisor: Ran Huo, R.Huo@tudelft.nl

Live cell imaging at nanoscale gives us insight into the cellular organization and function during dynamic process in life sciences. Super-resolution optical fluctuation imaging (SOFI) as a gentle live cell imaging approach has the potential not only for enhancing the resolution beyond diffraction limit, but also for extracting quantitative information of molecular density, which could help us better understand protein structure-function relationships in, for example, protein aggregation underlying neurodegenera-
tive diseases.

SOFI requires imaging data of fluctuating fluorescence signals that need to be tailored for optimal resolution, imaging speed, and quantitative readout. This project aims to evaluate and optimize novel classes of fluorescent probes and labeling techniques that are available at the Grußmayer lab, mainly self-labeling Halotags with exchangeable ligands (xHTLs) and self-blinking dyes, which also allows you to explore the possibility of multicolor live-cell imaging with SOFI.

Techniques

You will learn about mammalian cell culture and fluorescent labeling techniques. You will get hands-on experience with our state-of-the-art super-resolution microscopes and advanced image analysis. You will work closely with the interdisciplinary team at Grußmayer lab. Depending on your background and interests, you could either focus on optimization of data acquisition or data analysis.

Single-molecule super-resolution imaging of Huntington’s Disease

Further reading (click to link to article)

(Example) projects submitted by lab in past years

(2024-2025) Self-labelling Protein Tags for Quantitative Super-resolution Live-cell Imaging

Techniques

Further reading

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Confidants