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Additional correction images obtained at the time of image capture as in a posteriori correction are not available and therefore an ideal illumination model are used in priori correction. A priori method is therefore a preferred option. In digitizing images, the camera noise, background illumination intensity, and color temperature are the sources of image degradation to consider. Likewise you also have to consider the random noise due to uncorrelated fluctuations above and below, the fixed pattern noise characterized by pixel intensities consistently above random noise fluctuations. This is due to faulty CCD or pixel differences in charge leakage rate. Banding noise, which arises during the process of reading the data from the digital sensor or by interference with other electronic equipment are also included in camera noise.
The background illumination intensity provided by the microscope light source optics is commonly not homogeneous throughout the view field. This spectrum variation depends on the temperature of the filament. Therefore, images taken at different times may exhibit backgrounds with slightly different hues.
Another thing that affects the image quality is the color temperature of the light source. Light sources have a characteristic radiation spectrum. Color segmentation, color separation, hue quantification, etc. are procedures that are difficult to standardize by this.
Some microscopes have a button which sets a fixed voltage to the bulb to deliver fixed color temperatures. You can find this out by taking a series of background shots over a period of time and see whether the light distribution drifts over time.
The results become stable whenever you observe a plateau. If this is used, then the intensity of the light can be controlled with neutral density filters in the light path. The camera and microscope settings included make sure that it has an infrared filter; switch on all your equipment and leave it warm up for some time (the warming time depends on room temperature, how sensitive the equipment is to temperature and how long it takes to stabilize. You may want to do image capture during the plateau period.); switch off the camera auto gain; put the specimen on the stage, focus the objects, adjust the light/neutral density filters and condenser, set an appropriate camera exposure time. If the camera has a white balance function, you can apply it now on the illuminated bright field. Take out the specimen and apply the white balance; reposite the specimen and check again if the image histogram is not saturated and that the gray scale values span most of the gray scale space; and from now on, the settings of the camera and the microscope light should not be adjusted anymore.
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