The optical microscopy is one of the most powerful tool in the analysis of biological systems. The usual transmitted light microscope uses a white light lamp as source, what sometimes does not bring optimal results, making it necessary to introduce filters to change some illumination properties like the color temperature or the color itself. There is, of course, an intrinsic limitation on the use of filters that is the lack of an analogical control on the illumination properties and a practical limitation that depends on the number of available filters. To address this need, we developed an illumination system based on (Red, Green and Blue) RGB LEDs, were the microscope operator can control the intensity of each one independently and manually. This paper details the developed system and describes the methods used to compare quantitatively the images acquired while using the standard white light illumination and the images obtained with the developed system. To quantify the contrast, we calculated the relative population standard deviation for the intensities of each channel of the RGB image. This procedure allowed us to compare and understand the major advantages of the developed illumination system. All analysis methods have shown that a contrast enhancement can be obtained under the RGB LEDs light. The presented illumination allowed us to visualize the structures in different samples with a better contrast without the need of any additional optical filters.