Fluorescence imaging beyond intensity

Course overview
As fluorescence lifetime-based readouts become more accessible, this course aims to provide a broad understanding of how the lifetime-based information can be leveraged to solve biological questions. This application-centred course will provide insights into how this additional dimension of fluorescence information can help unveil functional and mechanistic information.

Audience
This course is aimed at PhD students and Post Docs with experience in fluorescence microscopy. It will enable them to better apply confocal fluorescence microscopy and in particular lifetime-based readouts to their cutting-edge research, disseminate their gained expertise to colleagues at their home institution and, eventually, to develop imaging methods beyond the state-of-the-art.

Modules/resources
Measuring protein-protein interactions
Environmental sensing with FRET-based and non-FRET-based biosensors
Using fluorescence lifetime for multiplexing
Increasing the resolution with tauSTED
Endogenous fluorescence – harnessing additional information and removing unwanted fluorescence like autofluorescence and background fluorescence
Sessions at the microscopes to practice using lifetime imaging in exemplary workflows
Analysis of the lifetime-based information and applicative conclusions
Learning outcomes
Understand the basics of fluorescence lifetime
Identify which types of research questions and approaches can benefit from lifetime-related measurements and how to design such experiments
Learn to use common biosensors, probes and fluorophores and understand the additional information and contrast provided by fluorescence lifetime
Understand the basis of endogenous fluorescence (in particular in tissue and animal models) and how fluorescence lifetime can help to exploit or cope with their signals
Acquire and analyze lifetime-related information in different experimental setups