In the present thesis, design and construction of fluorescent instrument via digital camera detector was carried out. A completely dark box, a LASER beam light, a cell-phone and an image processing Matlab-based software were used to build this instrument. Fluorescence image of solution was taken by cellphone’s camera and then transferred to image processing Matlab-based software to further analyzing and conversion into 3-dimensional data. Output response of image analyzer software is based on written algorithm in three matrixes: Red, Green and Blue. Obtained data from these matrixes were utilized for measuring the fluorescence and plotting calibration curves of Rhodamine B, Vitamin B 2 and Vitamin B 9 . The limit of detection, linear range and relative standard deviation (RSD) for rhodamine B were resulted in 1.06x10 -3 µg.lit -1 , 5x10 -3 -1µg.lit -1 and 4.8%, respectively. Same procedure was performed for Vitamin B 2 and had 4x10 -3 µg.lit -1 limit of detection, 2x10 -2 -6x10 -1 µg.lit -1 linear range and 3.9% RSD. Analyzing of Vitamin B 9 was lead to 3 µg.lit -1 limit of detection, 10-100 µg.lit -1 as linear range and 4.5% of RSD. Mentioned above compounds were analyzed again separately via Shimadzu Spectrofluorometer instrument and results from this two approach were compared with each other.