Carbon quantum dots (CQDs) are nanoobjects of a size below 10 nm. Due to their favorable features, such as tunable luminescence, unique optical properties, water solubility, and lack of cytotoxicity, they are willingly applied in biomedicine. They can be obtained via bottom-up and top-down methods. However, to increase their quantum yield they must undergo post-processing. The aim of the following research was to obtain a new type of CQDs modified with a rhodamine b derivative to enhance their fluorescence performance without biocompability deterioration. For their preparation glucose was used as a precursor and four different carbonizing agents which affected semi-and final products luminescence properties. The ready nanomaterials were investigated over their chemical structure by FTIR and NMR, whereas morphology was investigated by the TEM method. Their optical properties were determined by UV-VIS spectroscopy. Fluorescence behavior, photo-and pH-stability, as well as solvatochromism showed their applicability in various biomedical applications due to the controlled properties. The samples exhibited excellent antioxidant activity and lack of cytotoxicity on L929 mouse fibroblasts. The results showed that proposed strategy enables preparation of the superior nanomaterials with outstanding luminescence properties such as quantum yield up to 17% which can be successfully applied in cell labelling, bioimaging, and theranostics.Molecules 2020, 25, 736 2 of 18 can release cadmium ions leading to cell death. Importantly, it occurred that quantum dots may undergo certain modifications resulting in the change of their surface or chemical structure [27,28]. Finally, their preparation is expensive. Therefore, there was a need for a novel type of quantum dots which would be suitable for biomedical applications. In 2004, a new type of carbon-based materials was accidentally obtained during nanotube purification [29]. Until then, numerous types of carbon quantum dots (CQDs) have been obtained with various possible applications due to their very attractive properties, such as luminesce, good water solubility, high photostability, resistance to photobleaching or photoblinking, possibility, chemical inertness and, finally, lack of toxicity and biocompability [1][2][3]. Furthermore, CQDs' region of photoluminescence is very broad, starting in the ultraviolet and ending in the near-infrared. Although, until now, the luminescence behavior of carbon nanodots is still not fully clear, it is believed that crucial role plays two factors, namely, that the quantum size is below 10 nm and surface defects which may act as excitation energy traps. Interestingly, CQDs can successfully act as both electron donors and acceptors [2][3][4].CQDs can be prepared from various resources, such as peels (mango, orange, banana, pineapple, onion, seaweed etc.), meat, grains, nuts, or vegetable by-products, which makes them a low-cost material [30,31]. Currently, these nanomaterials are widely applied in drug delivery, gene delivery, biosensing, bioimag...