Principles of microscopy

This course is a broad overview of the concepts and theory behind microscopy. It is designed for people who are already doing some imaging but want to understand the basis of microscopy a little better and perhaps learn a bit about other types of microscopes/techniques. The course is . . .

  • Scheduled periodically - Next time January 7th and 8th 2010
  • Takes place as two 3-hour small group talks on different days
  • Open to everybody at Duke
  • Carries no credit
  • Has no practical content. If you want to learn how to use a particular microscope in the Light Microscopy Core Facility you can request training here.

 

I. How microscopes and confocals work

Introduction and overview
  • Modalities and configurations

Important concepts

  • Light and some very basic physics
  • Magnification and resolution
  • Numerical aperture

Optical components

  • Lamp, condenser, eyepieces
  • Objectives and aberrations
  • Dirt and dust

Transmitted Light

  • Kohler illumination
  • Contrast - Phase, DIC
 Fluorescence microscopes
  • Fluorescence
  • Fluorophores
  • How a fluorescence microscope works
  • Arc lamps
  • Filters/dichroics
  • CCD cameras
  • Stereoscopes
  • TIRF

Confocal microscopes

  • The confocal principle
  • How a confocal microscope works
  • Settings for a good image
  • Disk-scanning confocals
  • Two photon excitation

 

II. Samples, images and processing/analysis

Samples
  • Immunofluorescence
  • Live cell imaging
  • Fluorescent proteins
  • Photobleaching/toxicity
  • FRAP and Photoactivation/conversion
  • FRET reporters
 Images, processing and analysis
  • File storage and transfer
  • Files - formats, bit-depth, compression
  • Color
  • Useful software
  • Making figures
  • Presentation and processing in 3D
  • Principles of deconvolution
  • Measurements and quantification

If you are interested in this course please use this form to request enrollment

If you are graduate student and are interested in a more comprehensive course you could consider BIOLOGY 370: Principles and Practice of Microscopy.

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