Fundamental and Practical Applications of Light Microscopy in Biological Sciences

 High Resolution Imaging

 

Course Directors: Kent Keyser, PhD and Bradley Yoder, PhD

Text: Fundamentals of Light Microscopy and Electronic Imaging by Douglas Murphy and Michael Davidson

Purpose: The goal of the semester long course is to provide hands-on training in multiple forms of microscopy and digital imaging. The course will cover commonly utilized approaches in modern biological laboratories with emphasis on solid theory, design, practice, and data capture and analyses.

Short Description:

The course will provide GBS students and postdoctoral fellows with an in-depth background in the theory of modern microscopy analyses for researchers in the biomedical sciences complemented with hands-on practical exercises. The course will cover a wide range of fundamental and cutting edge approaches with training in experimental design and technical limitations, specimen preparation, diverse uses of bright-field, simple epifluorescence, single and multiphoton confocal, high resolution, live-cell, and intravital microscopy. The course will also provide training in specialized application such as particle tracking and co-localization, photo-activation, Ca2+ imaging, FRET, FRAP, FLIM, and TIRF, and methods for quantitative data analyses. The course will also cover image preparation for publication and ethical issues related to image manipulation.

Student Evaluation and Grading: Students’ mastery of the material will be evaluated through quizzes, class discussion and participation, and two tests that will consist of written and practical examination.

TOPICS TO BE COVERED

  • Overview of microscopy
  • FUNDAMENTALS OF LIGHT MICROSCOPY
  • LIGHT AND COLOR
  • ILLUMINATORS, FILTERS, AND THE ISOLATION OF SPECIFIC WAVELENGTHS
  • LENSES AND GEOMETRICAL OPTICS
  • DIFFRACTION AND INTERFERENCE IN IMAGE FORMATION
  • DIFFRACTION AND SPATIAL RESOLUTION
  • PHASE CONTRAST MICROSCOPY AND DARKFIELD MICROSCOPY
  • PROPERTIES OF POLARIZED LIGHT
  • POLARIZATION MICROSCOPY
  • DIFFERENTIAL INTERFERENCE CONTRAST MICROSCOPY AND MODULATION CONTRAST MICROSCOPY
  • FLUORESCENCE MICROSCOPY
  • TWO-PHOTON EXCITATION FLUORESCENCE MICROSCOPY
  • SUPERRESOLUTION IMAGING
  • IMAGING LIVING CELLS WITH THE MICROSCOPE
  • ADAPTIVE OPTICS (REVIEW ARTICLE
  • IN VIVO IMAGING
  • FUNDAMENTALS OF DIGITAL IMAGING
  • DIGITAL IMAGE PROCESSING