UvA Mastercourse Advanced Microscopy

Title: Advanced Microscopylogo-uva
Date: October (annual)
Location: FNWI, AMC & NKI
Contact: Mark Hink
Previous editions: 2011-2016

 

Objectives

The general aim of this course is to get hands-on experience and theoretical background in performing state-of-the-art advanced (light) microscopy applied to cell biology. Due to the revolution in genetic encoded labeling (GFPs, switchable probes, FlAsH and AGT-tagging, etc) and the revolution in advanced (light) microscopy (confocal, super-resolution, FRET, dynamics etc) it is now possible to image and quantify the dynamics of molecular processes inside cells with unprecedented resolution, speed and accuracy. Because the application of advanced microscopy is essential for modern research in (clinical) cell biology, this course provides training in the diverse modes of advanced microscopy. The microscope setups on which the training is provided, are part of the van Leeuwenhoek Centre for Advanced Microscopy (LCAM) in Amsterdam, in which the research groups of Molecular Cytology (Swammerdam Institute for Life Sciences, UvA), of Cell Biology and Histology of the Academic Medical Center (AMC) and the Cellular Biophysics Group, Dept of Cell Biology Netherlands Cancer Institute (NKI) collaborate and organize their advanced microscopy infrastructure.

micstudents

Contents

The course starts with a theoretical introduction in optical spectroscopy (fluorescence, absorption, anisotropy), microscope optics, fluorescent probes, image processing & data statistics. The theoretical courses are accompanied by hands-on practicals on advanced microscopy. For each day one microscopy technique is highlighted by giving the basic theoretical background with tutorials (morning) and hands on experience at the microscope (afternoon). After three weeks there will be a written exam. In the final week the students will work in small groups, focusing on specific technique of choice. Experiments will be performed to strengthen their knowledge and the results will be presented at the final day.

The topics that will be covered are:
◾Optics
◾Microscopy hardware
◾Fluorescence
◾Widefield microscopy
◾Confocal Laser Scanning Microscopy (CLSM)
◾Live cell microscopy
◾Spinning disk confocal microscopy
◾Total Internal Reflection Fluorescence Microscopy (TIRF)
◾Fluorescent dyes
◾Biosensors and optogenetics

◾Fluorescence Resonance Energy Transfer (FRET) microscopy
◾Filter- Ratio- and spectral FRET
◾FRET-FLIM

◾Fluorescence recovery after photobleaching (FRAP), fluorescence loss in photobleaching (FLIP) & photoactivation microscopy
◾Fluorescence lifetime imaging microscopy (FLIM)
◾Fluorescence correlation spectroscopy (FCS)
◾Super-resolution microscopy (STED, SIM, PALM, GS-DIM)
◾Electron Microscopy and tomography (EM)
◾Flow cytometry and cell sorting (FACS)
◾Data processing using ImageJ
◾Statistics for microscopy

 

Min/max participants

Maximum 25