15.Observation Make observations using darkfield microscopy

Make observations using darkfield microscopy

What is darkfield microscopy?

  • Darkfield microscopy is a method of using oblique illumination to observe unstained specimens.
  • Use high contrast to make light observation images visible on a dark background.
  • When using darkfield microscopy, the illumination source is focused in a circular shape by the diaphragm ring. The light passes through the edge of the condenser lens and illuminates the specimen obliquely.
  • By using a lens with a smaller numerical aperture than the condenser lens, only the scattered light in the specimen passes through the objective lens and will form a darkfield image.

Preparing for darkfield microscopy

  • The parts listed below can be used.
  • Darkfield slider
  • Objectives lenses for darkfield microscopy
    • The objective lenses that can be used with darkfield microscopy have restrictions such as the ones listed below.
    • When using darkfield microscopy
      NA0.65 or less
    • When making observations in the light field position
      NA 1.25 or less

Make observations using darkfield microscopy

  • Raise the stage, remove the condenser and the dummy slider in it.
  • After inserting the darkfield slider, return the condenser, and adjust the position with the condenser’s vertical movement knob.
  • Place the darkfield position in the light path, set the open aperture lever to the same numeric value as the objective lens, and bring it into focus.
  • After adjusting stage height with the coarse adjustment knob, bring it into focus with the fine adjustment knob and observe.
  • Return to light field.
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