Currently, there are two common methods to acquire the TLC plate image. These methods use a flatbed scanner or employ a charge-coupled device (CCD) camera. TLC imaging using chargecoupled device (CCD) detectors has been shown to enable rapid and efficient analysis because entire TLC plate can be imaged in a single exposure, yielding a rapid quantification of the chromatographic spots. For imaging system, in addition to digital scanning, image acquisition by digital camera has also been successfully used for TLC in case of fluorescent compounds. In addition, due to the advantages of the uniform lighting of surface, short time of scanning and high optical resolution, different flatbed scanner systems have been employed in the TLC-quantitative evaluation of various classes of compounds and in some cases the results were compared to those obtained with the use of slit-scanning densitometry. Moreover, taking into account the advantages of chemometrics, Hemateenejad et all. Have successfully applied multivariate analysis to the images of TLC plats for simultaneous determination of compounds even in the presence of overlapped spots.