Neurizons brings to you method workshops, for state of the art and emerging techniques in Neuroscience. It is an opportunity to get hands on practical training and an introduction to the new trends in scientific methods. Experience the work atmosphere and learn first hand the techniques you read about and always wanted to try.
1. Expansion Microscopy
Offered by: Dr. Sven Truckenbrodt (S.O. Rizzoli Lab)
Number of participants: 5
Duration: 4 hours
Expansion Microscopy (ExM) is the most recent addition to the biologist’s toolkit for super-resolution imaging. All super-resolution imaging techniques used so far (e.g. STED, PALM/STORM) circumvented Abbe’s resolution limit by separating fluorophores in time, by detecting neighboring fluorophores separately one after another. ExM instead separates fluorophores in space, by increasing the physical distance between neighboring fluorophores. This is achieved by embedding the sample in a swellable gel matrix after immunostaining. The gel matrix can then be expanded equally in all three dimensions to obtain a resolution of at least ~80 nm, and up to ~25 nm (depending on the ExM technique used).
ExM has undergone rapid development since its introduction in 2015 (Chen et al., 2015). It can now be used with conventional antibodies (Chozinsky et al., 2016; Tillberg et al., 2016), and obtains a resolution that rivals STED and PALM/STORM (Chang et al., 2017). ExM has the following advantages over other super-resolution techniques:
- No specialized instrumentation required, a resolution of up to ~25 nm can be obtained on conventional epifluorescence microscopes
- Multi-colour super-resolution imaging in 3-4 channels is possible without any additional effort, with almost identical resolution in all channels
- ExM can be performed on any conventional immunostaining (with the restriction that not all fluorophores are compatible with ExM)
We will perform ExM on exemplary samples provided during the course: COS7 cells (Tubulin), neuron culture (SVs, AZ, PSD), and rat brain slices.
2. FlyGym (Tracking Drosophila optomotor responses)
Offered by: Sebastian Molina Obando, Madhura Ketkar (M. Silies Lab)
Number of participants: 7-8
Duration: 4 hours
With a variety of sophisticated tools available for its genetic manipulation, the fruit fly Drosophila Melanogaster offers many advantages as a model organism. Combining genetic manipulations in specific neuronal subsets together with behavioural assays can help us to address a wide range of neuroscientific questions. The aim of the workshop is to present how an animal’s locomotion can be tracked during behavioural experiments. We will track the responses of the Drosophila during visual stimulation in a virtual reality setup. In the first part (~1.5 hours) of the workshop we will discuss the theory behind this approach and explain how measurements acquired from a fly walking on an air-supported ball can be used to compute the fly’s walking velocity. Afterwards, participants will have a ‘hands-on’ experience with our behavioral setup, and will be guided through a real experiment, including analysis and visualisation of the results. Participants will also have a chance to learn how these experiments are designed in our lab to efficiently address our scientific interests.