Combination of fluorescent microscopy and phase contrast microscopy- while a study of a specimen can be done with a fluorescent microscope or with a phase contrast and still obtain a good result, surely a combination of the two methods would more than double the expected results in terms of clarity and vividness. A primer of epifluorescence microscopy: The fluorescent microscope is used to visualize specimens that fluoresce, that is to say, emit light of a different color, always a higher wavelength, than the light absorbed by the specimen. Fluorescence occurs either because of naturally occurring fluorescent materials in the cells, such as chlorophyll and related molecules or because the cells have been stained with a fluorochrome. Fluorochromes are stains similar to cell and tissues stains used in light microscopy and have been chosen or designed to be highly specific in their attachment to molecules in cells.

The use of fluochromes has made it possible, to view cells and sub-cellular components of cells with a high degree of specificity amid non-fluorescing material. There may be times when it is useful to locate and view a specimen using phase contrast and then add fluorescence illumination to view an organelle or the location of a particular molecule in the cell. Once the specimen, say a cell, is located under phase contrast, the light intensity knob is used to turn down the halogen lamp and the fluorescence shutter is set to the on position. The correct filter cube is placed in the light path and the intensity of the halogen lamp is adjusted so that the cell can be seen with phase and the fluorescing structure s is visible. The applications of phase contrast and fluorescent microscopy- living cells and most cell organelles are often difficult if not impossible to see by brightfield microscopy because they do not absorb, refract or diffract sufficient light to contrast with the surrounding medium. The phase contrast microscope was developed to improve contrast differences between cells and organelles and the surrounding medium, making it possible to see cells and organelles without staining.

The technique used is based on the fact that cells differ in refractive index from their surroundings and thus bend some of the light rays passing through them. Light rays passing through a transparent specimen, most unstained cells, are transparent, emerge as either direct rays unaffected by passage through the specimen unaltered in intensity and phase or diffracted rays bent as they pass through the specimen, altered intensity or phase. This effect is amplified by the phase annulus coupled with special phase rings in the objectives leading to a dark image on a light background. The fluorescent microscope is used to visualize specimens that fluoresce, that is, emit light of a different color, always a higher wavelength than the light absorbed by the specimen. Fluorescence occurs after light is absorbed either because of naturally occurring fluorescent materials in the cells such as chlorophyll and related molecules or because the cells have been stained with a fluorochrome. Fluorochromes are stains similar to cell and tissues stains used in light microscopy and have been chosen or designed to be highly specific in their attachment to molecules in cells. The use of fluochromes has made it possible to identify cells and sub-cellular components of cells with a high degree of specificity amid non-fluorescing material. An Olympus microscope is equipped with 3 filter cubes FITC, TRITC, and DAPI named for the common dyes they are used with.