Custom-built wide-field epi-fluorescence microscope setup with two solid-state lasers with 640 nm, and 488 nm (MLDTM, Cobolt). Acoustic-optical tunable filter (AOTF; AA Optoelectronic) is used to control the intensity of the individual laser lines for excitation. The collimated laser beams are introduced into the microscope through an achromatic focusing lens, providing widefield Köhler illumination of the sample. The fluorescence images are recorded using an inverted IX71 microscope (Olympus) with a different type of objective lens and illumination condition (TIRF or low angel). The fluorescence emitted from the sample is collected again by the same objective lens, and separated from the laser excitation light using a dichroic mirror and an emission bandpass filter obtained from (Semrock). Detection is done by two Andor iXon EMCCD cameras connected through TuCman image splitter enabling us to acquire fluorescent images from two different wavelength religions simultaneously. The z-axis positions are controlled by E-665 PZT-amplifier/servo (PI). The image acquisition is done using IQ3 from Andor. The setup is equipped with variety of accuracies that enables us to measure live cell behavior in micro fluidics.