791014 Advanced light and chemical microscopy in life science (in Eng.)

Vorlesung und Übung
Vortragende/r (Mitwirkende/r)
Schosserer, Markus , Debreczeny, Monika
Institut für Biotechnologie
Angeboten im Semester
Wintersemester 2017/18
Unterrichts-/ Lehrsprachen


Basic Light Microscopy:
-) Types of microscopes (upright, inverse, Stereomicroscope, Macroscope)
-) Components of the visible light path
-) Contrast methods: Phase Contrast, Differential Interference Contrast (DIC), -) Polarisation – Comparison of the different methods
-) Köhler Illumination
-) Numerical Aperture
-) Diffraction limit
-) PSF

Fluorescent Microscopy, Immuncytochemistry and FISH:
-) Components of the fluorescent light path
-) Digital detectors and their applications
-) Basic Principles of Immunocytochemistry
-) Fixation techniques
-) Dyes
-) Basic Principles of FISH
-) Immunocytochemistry and FISH of Tissue Sections
-) Examples

Microscopy of thick specimen/Imaging in 3D:
-) Confocal Microscopy – Spot Scanning and Spinning Disc
-) Two-Photon Microscopy
-) Deconvolution
-) Light-Sheet Microscopy
-) Comparison of the different methods - examples

Life-Cell Imaging:
-) Life-Cell Imaging dyes
-) Fluorescent proteins
-) Photo-switchable fluorescent proteins
-) Phototoxicity - Points to consider to keep cells alive and happy
-) Examples

Super-Resolution Microscopy:
-) Going beyond the diffraction limit
-) Pointillist imaging: PALM and STORM
-) SIM
-) Comparison of the different methods - examples

Digital Image Processing and Chemical Microscopy:
-) Use of ImageJ for quantification of fluorescence intensities and particle counting
-) Deconvolution (using Huygens)
-) Raman Microscopy

Inhaltliche Voraussetzungen (erwartete Kenntnisse)

Basic knowledge of cell biology strongly recommended! This course is specifically recommended for PhD-students.


Students will know the basic principles and applications of microscopy in life sciences and have confidence in handling basic light and fluorescent microscopes. They will be able to select the appropriate microscope and design an experiment including the proper controls for their individual application. Furthermore, students will be able to identify important aspects of the different techniques, critically interpret results and perform basic troubleshooting. They will understand the basic principles and applications of more advanced microscopical techniques and thereby have the confidence to gain a deeper understanding by consulting the relevant literature on their own.
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