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How to set up a microscope for köhler illumination should be an ingrained knowledge for every microscope user and should always be the first step of action once you switched on your microscope.

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In this pdf you will find a step by step guide to properly set up your microscope for Köhler illumination and thus achieve optimal imaging results:

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科勒照明

In the case of microscope design, the optical engineer’s goal should be uniform illumination across the whole sample. Uniform illumination has the additional advantage of creating higher contrast. One way to create such illumination is by using what is called a Koehler illumination system. To create a Koehler illumination setup, we need two adjustable diaphragms: one near the light source (also called condenser diaphragm or CD) and the other close to the sample (also called field diaphragm).

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With over 20 years of experience and 800+ unique optical systems designed, Optics for Hire specializes in advanced optical engineering. If it uses light, we've worked on it.

When designing any optical system, an optical engineer needs to consider how illumination will be used: specifically, how the illumination could affect the way the image is created. If not carefully designed, some illumination systems may create a kind of vignetting where the illumination will have a higher intensity at the center while decreasing to zero around the edges. Alternatively-as in the case of simple microscopes that are illuminated by halogen bulbs – the microscope can create an image of the lightbulb itself.

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August  Köhler, who first introduced the Köhler illumination procedure, was  part of the Carl Zeiss corporation. Here you can read how Zeiss  Microscopy explains Köhler illumination.

Because the light source is not focused at the sample, the light behaves as an extended diffused source. The combination of the apertures of both diaphragms determines the numerical aperture of the microscope system. As the CD is increased, the NA also increases thus creating higher resolution on the system.

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Criticalillumination

By adjusting the condenser diaphragm, the optical engineer can modify the sample contrast and depth of field. In addition, we can control the amount of light reaching the sample. The collector lens projects a larger image of the lamp filament where the field diaphragm is placed. By controlling the CD’s aperture, we can control the angular aperture of the light cone. The field condenser (FD) can control the illumination area on the sample.

With over 15 years of experience and 500+ unique optical systems designed, Optics for Hire specializes in advanced optical engineering. If it uses light, we've worked on it.

In this 12min video, Ron Vale talks you through a step-by-step procedure of aligning the lamp and condenser to achieve Koehler illumination.

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The most fundamental, but often forgotten step towards a successfull microscopy session is the proper alignment of your microscope. In order to achive optimum system performance and acquire consistent, interpretable image data the illumination of the specimen must be adjusted to the most optimal, even level.