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Light Microscopy

Light Microscopy. Uses Before you go to the EM Quick check Maybe all you need Hydrated and living materials. Light Microscopy. Systems Parts Illumination Sample stage Imaging. Light Microscopy. Parts of the light microscope Stand Illuminator Stage Objective(s) Tube Eyepiece(s)

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Light Microscopy

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  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)

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