This chapter explores the role of optical fluorescence techniques in studying pictorial binders and varnishes, highlighting both their strengths and limitations. It emphasizes the noninvasive and in situ nature of these methods, which makes them particularly valuable in art conservation. Binders and varnishes, composed primarily of organic molecules, often exhibit intense optical emission. This property is widely used in conservation to detect the presence of these materials, along with other organic fluorescent substances, on the surfaces of paintings. However, identifying their chemical composition through fluorescence is far more challenging. This difficulty arises because binders and varnishes typically contain multiple fluorophores, each with unique optical emission properties. The result is a broad, spectrally complex fluorescence signal that lacks specificity. Additionally, the fluorescence characteristics of these materials can change significantly over time due to aging and degradation processes, further complicating identification. Factors such as chemical interactions with surrounding molecules and reabsorption effects add another layer of complexity. To address all these challenges, advanced molecular fluorescence spectroscopy techniques are required. Approaches using multiple excitation wavelengths and lifetime-sensitive measurements are better suited to distinguishing between different binders and varnishes or, at a minimum, categorizing their chemical class. Through examples of fluorescence studies on binders and varnishes from the last 50 years, it is clarified how molecular fluorescence can offer a valuable in situ, noninvasive approach for analyzing binders and varnishes in artworks, provided that fluorescence-based methods with high molecular specificity are employed. Further, it is clarified how these techniques are most effective when integrated into a multi-analytical strategy that combines complementary spectroscopy methods, which remains the preferred way to assess the composition and nature of binders and varnishes in artworks.