A. Prepare the Microscope and Camera
1. Set up Köhler illumination. (More on Setting Kohler illumination)
2. Adjust the transmitted illuminator’s voltage to 9 - 12 volts, or to the manufactures preset for imaging.
4.. Remove all colored filters from the transmitted light path.
5.. Set the camera system to the resolution at which you wish to make your photographs.
6. Image an empty field by transmitted light, turn off any gain or offset settings, adjust the camera’s exposure setting to give a neutral gray field, and perform white balance.
B. Prepare your Slides
1. Clean your slides on both sides.
2. Try to insure that only one cover glass is present.
3. Place the slide on the microscope stage with the specimen side facing the objective lens.
C. Make Final Adjustments and Shoot
1. Adjust Köhler illumination for the objective lens in use. You can stop down the field iris to just outside the area that will be in the photograph; you should carefully adjust the condenser iris to achieve the best contrast without creating diffraction artifacts.
2. Focus: at magnifications below 10 X use a focus aid or magnifier if one is present. If using a high dry objective lens with a cover glass correction collar, adjust the collar for the sharpest possible image with the condenser iris fully open, then re-close the iris. If you are using an oil immersion lens, check for bubbles by observing the objective’s back focal plane.
3. Adjust the exposure time using neutral density filters and the camera’s exposure controls to achieve a relatively short exposure - between 1/10 and 1/100 second.
4. Recheck the focus by observing the image on your monitor, allow the microscope to settle, and capture the image.
5. Save the image as an uncompressed TIF image file.