Modern research in molecular, cellular, and developmental biology requires the precise measurement of cellular or subcellular activity in two- or three-dimensions. Many available fluorescence microscope techniques are yielding limited detail about the organization and dynamics of complex cellular structures. But the multi-photon (two- or more photon) excitation fluorescence imaging microscopy (MEFIM) system proposed here will provide a unique, state-of-the-art opportunity to integrate advances in optical microscopy, low- light video detection, and two- or three-dimensional image analysis in measuring the fluorescent signals from living cells. The MEFIM system uses longer excitation wavelength, which increases the penetration of the excitation of the sample, yet essentially reduces photobleaching and autofluorescence. But, more importantly, the infrared light excitation in the MEFIM techniques allows maintaining cell viability for longer periods of time and thus acquisition of more images. In our current work, we integrated the MEFIM system with one-photon laser scanning confocal microscope coupled to a verdi pumped tunable femtosecond pulsed ti- sapphire laser. Appropriate wavelength was tuned to excite the fluorescently labeled specimens and MEFIM images were acquired from living biological specimens at different optical sections. The importance of the MEFIM image acquisition and processing for biomedical sciences are discussed.