Defocusing effects in modulation transfer function (MTF) measurement of charge coupled device (CCD) cameras is the main focus of current paper. We introduce measuring Point-spread function (PSF) in order to calculate the MTF and further more we will study the shape of MTF and its cut-off frequency by adjusting the lens focusing in different locations. A collimated white light LED by broadband spectra is used as source and our results shows that the cut-off frequency is related to defocusing.