The Counter Reformation period in European art was a process in which some themes were transformed and also new themes emerged in Christian iconography. While the Catholic Church attempted to respond to the Protestant Reformation by using religious art as propaganda on the one hand, it focused on penitence – which was strongly criticized by Luther - on the other hand by concentrating on the sacraments which are determinative in terms of the Catholic doctrine. The most significant reflection of penitence in religious painting, which is more effectively underlined by the decrees of Council of Trent (1545-1563), is indisputably the figure of Mary Magdalene, with her biography evolving from sinfulness to penitence. The biography of the saint, whose controversial identity in the canonical gospels was “clarified” by the homily of Pope Gregory, was expanded with narratives in the late medieval literature that detailed the period of her seclusion. Thus, the image of the Penitent Mary Magdalene, distinguished by being the first follower to witness the resurrection of Jesus Christ, served as a remarkable model in the defence of the Counter-Reformation. Presented by the Catholic Church as a symbol of repentance to both Protestants and Christians in general, Mary Magdalene has been depicted in paintings with the characteristics of portraits in which her nudity is often emphasized with reference to her sinfulness since the beginning of the 16th century. Tiziano’s painting dated to 1531 (Palazzo Pitti, Florence) is one of the prototype examples of such depictions in European art. With the influence of the spiritual environment that emerged with the Counter-Reformation and intensified in the Baroque period, this “erotic portrait” brought a new mode of representation composed with the visual motifs of contemplation on repentance and death while renunciation of worldly things and the idea of memento mori became the main statement of these scenes. As a matter of fact, in the widespread depictions within this period, the skull motif can be defined as an invariable attribute. In the study, the content of the saint’s seclusion was examined in the context of both gestures of the figure and the visual motifs included in the composition, with reference to painting series of the Penitent Mary Magdalene by Tiziano, El Greco, Domenico Fetti and Georges de la Tour. The basis of this preference lies in the idea that motifs such as a skull, the Bible, a candle, a mirror, and jewels allow different meanings to be read in the scene created by these painters. While the paintings of Tiziano and El Greco trace the contrast in the sinful/penitent identity of the saint, Domenico Fetti and Georges de la Tour deepened the contemplation of the saint with “a melancholic touch” by feeding off the repertoire of the Baroque allegories. This paper aims to analyse the visual code revealed in the iconography of the Penitent Mary Magdalene, based on Panofsky’s “a world of symbolic values” paradigm, within the framework of the spirit of the Counter-Reformation, religious literature, Christian symbolism, the new mode of piety and the melancholic temperament.