Molecular Microscopy (B-KUL-H08G4A)
This course in taught at Chalmers Tekniska Högskola.
More information: www.student.chalmers.se/sp/course?course_id=17402
The course provides theoretical as well as practical know-how in modern light microscopy, inviting the student to the nano-scale worlds of physics, chemistry, bio- and material sciences. An overview over the different light-matter interaction mechanisms in the focal volume will be given, and how this information is passed on as an image from the micro- to the macro-world by an optical system. In particular, we will address how cell structures and single bio-molecules can be selectively imaged using various microscopy techniques based on fluorescence and advanced spectroscopy.
Basic courses in physics are recommended, e.g. Experimental Physics.
Is also included in other courses
The topics of the course are:
1. The theory of image formation
a. Optics, light paths and image formation in a microscope
b. Diffraction, Fourier optics and spatial filtering
c. Definitions and improvements of image quality
d. Introduction to image analysis
2. Three dimensional microscopy: wide-field/deconvolution, confocal, structured illumination and non-linear microscopy
3. The Microscopy Workshop - microscopy in practice
a. The anatomy of a modern microscope: the optics, different objectives, polarizers, filters and the DIC prism
b. The anatomy of a modern microscope: from light sources to detectors
c. Hands-on training: Koehler illumination, wide-field, dark-field, polarization, DIC and fluorescence microscopy
4. The physics in the microscope probe volume
a. Visualizing the invisible - contrast mechanisms: absorption, refractive index, polarization
b. Molecular microscopy: fluorescence, vibrational and non-linear effects
5. Fluorescence microscopy in contemporary cell biology: sample preparation, fluorescent markers such as GFP and emerging alternatives, applications also including advanced techniques such as FLIM and FRET.
6. Research projects at the Centre for BioPhysical Imaging, one of the following:
a. Single molecule detection by means of TIRF microscopy
b. Fluorescence lifetime imaging - FLIM
c. CARS - Coherent anti-Stokes Raman scattering microscopy
d. Surface enhanced Raman (SERS) microscopy
e. Surface plasmon resonance and nano-particle sensors
f. Optical manipulation
7. Oral presentations of the research project, including scientific discussions
8. High sensitivity microscopy: surface plasmon effects and SERS microscopy, single molecule detection
9. Optical manipulation: optical tweezers, manipulation, SLM's
Handbook of Biological Confocal Microscopy (Kluwer Academic Publishers), James B. Pawley (Editor), 2006
Introduction to Optical Microscopy, Digital Imaging, and Photomicrography: at a website