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Fluorescently tagged proteins are commonly used to tag different cell types within heterogeneous tissues. However, fluorescently labeled cells vary in their optical properties and fluorescence intensity. To evaluate the effects of differences in fluorescence intensity on optical signal, we performed two-photon excitation fluorescence microscopy on tissue sections labeled with fluorescein isothiocyanate (FITC) and 7-amino-4-methylcoumarin (AMC). FITC and AMC fluorescence images of the same region showed distinct differences in their color and contrast. A bright-field image of the region of interest showed that the cellular density varied considerably between the two types of cells. The average fluorescence intensity for each cell type was quantified using a region of interest. No significant changes in the average fluorescence intensity were observed within the boundaries of the region of interest for AMC-labeled cells. However, significant differences were observed in the average fluorescence intensity for FITC-labeled cells. The FITC-labeled cells showed a significantly lower average fluorescence intensity than AMC-labeled cells. These