790014 Advanced light and chemical microscopy in life science (in Eng.)


Art
Vorlesung und Übung
Semesterstunden
2
Vortragende/r (Mitwirkende/r)
Debreczeny, Monika
Organisation
Angeboten im Semester
Wintersemester 2023/24
Unterrichts-/ Lehrsprachen
Englisch

Lehrinhalt

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
-) Köhler illumination
-) Numerical aperture (NA)
-) Diffraction limit
-) Point spread function (PSF)

Fluorescence microscopy, immunocytochemistry, and FISH:
-) Components of the fluorescence light path
-) Digital detectors and their applications
-) Basic principles of immunocytochemistry
-) Fixation techniques
-) Fluorescent dyes
-) Basic principles of FISH

Microscopy of thick specimen/imaging in 3D:
-) Confocal microscopy – spot scanning and spinning disk
-) Two-photon Microscopy
-) Deconvolution
-) Light-sheet microscopy

Life-cell imaging:
-) Life-cell imaging dyes
-) Fluorescent proteins
-) Photo-switchable fluorescent proteins
-) FRAP, FLIP, and FRET
-) Phototoxicity

Super-resolution microscopy:
-) Going beyond the diffraction limit
-) Pointillist imaging: PALM and STORM
-) STED
-) SIM

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 is strongly recommended! This course is specifically recommended for PhD-students.

Lehrziel

Students will know the basic principles and applications of microscopy in life sciences and will have increasing 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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