Single-molecule localization microscopy and single-particle tracking (SMLM and SPT)

Seeing is believing. To understand the brain and nervous system, we need to know how mechanisms are mediated at the subcellular, cellular, and network levels. Regulation of neuronal communication at synapses is an example of a complex response at the subcellular level, with impacts on neuronal  network properties that in turn support functions from visual processing and pain regulation, to motor control, memory formation and emotion. Cellular signaling, homeostasis and change rely on proteins.  In neurons, we need to be able to observe protein dynamics (activity, localization, molecular interactions) within the tiny compartment of the synapse, as they occur in the context of neuronal activity and synaptic signaling.

NORBRAIN UiB will provide three complementary platforms for nanoscale optical interrogation of neuronal and synaptic function. The platforms will enable: 1) super-resolution-based localization and tracking of protein movement in cell culture preparations, 2) imaging of protein-protein interactions in intact tissue such as rodent brain slices, and 3) structural and functional imaging of single neurons and networks in intact neural tissue, in combination with neurotransmitter uncaging to study structural and functional plasticity and dendritic integration. 

Single-molecule localization microscopy and single-particle tracking (SMLM and SPT)  

A widefield microscope will have state-of-the-art capabilities for intracellular localization and tracking of proteins or RNA, and detection of cell surface diffusion and membrane trafficking events, at the single-particle level. This instrument is suitable for use in primary cell cultures, stems cells, cell lines, or simliar in vitro preparations.  

The instrument can used for super-resolution imaging using photoactivation localization microscopy (PALM), point accumulation in nanoscale topography (PAINT), total Internal reflection fluorescence (TIRF) microscopy, and quantum dot imaging.  

The University of Bergen (UiB) node is a cellular and molecular neuroscience unit with Clive Bramham and Espen Hartveit as local coordinators. 

The equipment is planned for installation and testing in 2021, and will become available to qualified users nationwide. A practical training course will be offered on a regular basis.