Köhler illumination is a sophisticated method employed in microscopy to enhance specimen lighting by precisely focusing the light source at the level of the condenser diaphragm. When the condenser is accurately positioned relative to the specimen, this technique significantly improves image clarity and detail, ensuring that microscopic observations are of the highest quality.
Process of Setting Up Köhler Illumination
Required Materials:
- A specimen slide (to be focused under 10× magnification)
- A compound microscope
Steps to Achieve Köhler Illumination
- Place the specimen slide on the microscope stage and focus using 10× magnification.
- Completely close the iris diaphragm.
- If the light source is not centered, adjust it using the condenser centering screws until the light is centered.
- Utilize the condenser adjustment knobs to raise or lower the condenser until the field edges become sharp (refer to Figures 1 and 2).
- Gradually open the iris diaphragm until the entire field of view is evenly illuminated.
When to Set or Check Köhler Illumination
- During routine microscope maintenance
- After the microscope has been moved or transported
- Whenever the sharpness of the viewed objects seems suboptimal