790014 Advanced light and chemical microscopy in life science

Type
Lecture and exercise
Semester hours
2
Lecturer (assistant)
Debreczeny, Monika
Organisation
Offered in
Wintersemester 2023/24
Languages of instruction
Englisch
Content

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, dark field illumination,
-) Köhler illumination
-) Objectives and Numerical aperture (NA)
-) Diffraction limit
-) Point spread function (PSF)

Wide-field Fluorescence microscopy:
-) Components of the fluorescence light path
-) Digital detectors and their applications
-) Basic principles of immunocytochemistry
-) Fixation techniques
-) Fluorescent dyes
-) Life-cell imaging dyes
-) Fluorescent proteins
-) Photo-switchable fluorescent proteins
-) FRAP, FLIP, and FRET

Microscopy of thick specimen/imaging in 3D:
-) Confocal microscopy – hardware elements of a confocal laser scanning system
-) Spinning disk Confocal microscopy
-) Two-photon Microscopy
-) Light-sheet microscopy

Super-resolution microscopy:
-) Going beyond the diffraction limit
-) Pointillist imaging: PALM and STORM
-) Stimulated emission depletion (STED)
-) Structured illumination microscopy (SIM)
-) semi-superresolution, various tricks to see subtle details

Label-free imaging, chemical microspectrometry
-) Fourier-transform Infrared spectroscopy/microscopy-)
-) Raman microscopy
How to publish images acquired with a microscope
Previous knowledge expected

Basic knowledge in cell biology is recommended, not obligatory. This course is specifically recommended for PhD-students who plan to acquire microscopy images from their object of interest (bacteria, yeast, mammalian cells, tissue sections, non-biological materials, environmental samples, ...).
Objective (expected results of study and acquired competences)

Students will learn 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.
You can find more details like the schedule or information about exams on the course-page in BOKUonline.