Spring Semester

Description: This course provides biologists, material scientists, and students from other disciplines with the theoretical background and practical techniques of sample preparation, operation of light microscopes, image acquisition and image processing. In addition, students will receive individual instruction, which will facilitate the completion their later research involving LM.

Course Format: Lectures: Wednesday and Friday 1-2 PM. Lab session: Wednesday 2-5 PM. Laboratory sessions will permit the students to receive formal education in the basic theory and operation of research light microscopes, including a confocal microscope,  in different observation modes. Following the formal theory and practice sessions, each student will be required to pass a "check-out" test which allows the student to independently complete their own research projects using the LM. A written exam will cover the topics presented in lecture. The research project will be presented during the final class session.

Homework: Each student is required to prepare a poster presentation describing microscopic analysis of a specimen examined in the LM during the course. The presentation should be comprised of three sections: 1) Introduction, 2) Materials & Methods and 3) Results & Discussion (including presentations of micrographs with annotations, figure legends, and image interpretation).

Lecture Topics:

• Theories of light
• The Light Microscope: history and evolution, components of a compound microscope  
• Theory of image formation, diffraction-limited resolution
• Optical aberrations and their correction
• Illumination and image forming techniques:
  • Brightfield Optics
  • Oblique Illumination Techniques
  • Imaging of Phase Objects
  • Polarization Optics
  • Fluorescence Microscopy
  • Confocal Microscopy
• Non-linear microscopy: Multi-photon excitation, Second Harmonics Imaging
• Overcoming difraction limits - superresolution light microscopy techniques
• Image recording, processing, and analysis

Coordinator: Dr. Andreas Holzenburg

Instructors: Dr. Stanislav Vitha, Dr. Karl Aufderheide, Dr. Alvin Yeh