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Light Microscopy

Light Microscopy

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Light Microscopy

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Presentation Transcript

  1. Light Microscopy • Uses • Before you go to the EM • Quick check • Maybe all you need • Hydrated and living materials

  2. Light Microscopy • Systems • Parts • Illumination • Sample stage • Imaging

  3. Light Microscopy • Parts of the light microscope • Stand • Illuminator • Stage • Objective(s) • Tube • Eyepiece(s) • Eye, camera

  4. Light Microscopy • Illumination • Through the sample • Incident on surface of sample

  5. Light Microscopy • Through sample illumination • Kohler illumination • Open field and condenser diaphragm • Focus sample • Close field diaphragm and focus with condenser focus control. Use centering controls to center image. • Open field diaphragm to fill image • Take out eyepiece and adjust condenser aperture to ¾ fill BFP of objective

  6. Light Microscopy • Incident Light illumination • Use special objectives • Almost setup for Kohler illumination • Objectives serve as condenser too

  7. Light Microscopy • Objectives • Achromats- RB chromatic and G spherical corrected • Semi-apochromats- RBG chromatic + BG spherical • Apochromats-RBBG chromatic + BG spherical • “Plan” objectives correct for field curvature

  8. Light Microscopy • Special Techniques • Darkfield (hollow cone illumination)

  9. Light Microscopy • Special Techniques • Phase contrast (blocking phase object interactions with illumination)

  10. Light Microscopy • Special Techniques • Polarized light (polarized light interaction with birefringent materials)

  11. Light Microscopy • Special Techniques • DIC (Nomarski) (shearing interferometry)