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Principles of Microscopy and Imaging

Intensive training course/workshop. Offered once per year (See Training Overview). Training through lectures, bench work and hands-on experience at different imaging workstations. Discussions about appropriate sample preparations, analysis and image acquisition. Critical interpretation of data, and discussion of research applications. Multidisciplinary and training. Invited speakers.

Topics of "Principles of Microscopy and Imaging":
1) Overview on Microscopy and Imaging.


o An introduction to the history and development of microscopy with particular emphasis on principals and techniques relevant to the biosciences.

Principles of Wave Optics

o Duality of light (superposition),Newton and Huygens
o Image formation, light paths, magnification, scale, diffraction, wavelength
o Discussion of resolution (Raleigh’s Criterion), contrast, intensity information and phase information

Principles of Microscopy

o General
• Lenses, aberrations and corrections
• Standard Nomenclature
• Parts of a microscope
• Light sources and color temperature

o Transmitted Light Techniques
• Brightfield (Koehler Illumination)
• Phase Contrast (Cernike)
• Darkfield
• Polarization
• Differential Interference Contrast (Nomarski)

o Reflected Light Techniques
• Fluorescence
• Principals (Stokes)
• Light Sources
• Filters
• Fluorochromes


2) Imaging and Documentation.

o Photography
• Film speed (DIN)
• Film types
• Focus and exposure
• Issues with color film vs. B&W
• Adaptors and vignetting

o Digital Imaging
• Information Management
• Documentation and digital technology Chips, resolution, sensitivity and adaptors
• Professional versus consumer cameras
• Cooling and noise
• Nomenclature and technology
• File types and compression

3) Live cell imaging.

o Temperature control
• Thermal stability, mass, and expansion
• Stages and Environmental enclosures
• Focus and movement

o pH control
• Methods and requirements
• Controllers
• Custom chambers
• Time Lapse

4) Fluorescence Imaging and Microscopy.

o Methods and requirements
• Choice of filters
• Applications
• FISH (fluorescent in situ hybridization)
• Overview on other fluorescent applications

Automated sample analyses.
o Screening Technologies
• Rare Cell Detection
• Metaphase Finding
• Spot counting ©2002