Development of Red-Emissive Porphyrin Graphene Quantum Dots (PGQDs) for Biological Cell-Labeling Applications

Red and near-infrared emission is a highly desirable feature for fluorescent nanoparticles in biological applications mainly due to longer wavelengths more easily being able to deeply penetrate tissues, organs, skin, and other organic components, while less autofluorescence interference would be produced. Additionally, graphene quantum dots (GQDs) that contain unique optical and electrical features have been targeted for their use in cell labeling applications as well as environmental analysis. Their most desirable features come in the form of low toxicity and biocompatibility; however, GQDs are frequently reported to have blue or green emission light and not the more advantageous red/NIR emission light. Furthermore, porphyrins are a subgroup of heterocyclic macrocycle organic compounds that are also naturally occurring pigments in nature that already contain the desired red-emission fluorescence. Therefore, porphyrins have been used previously to synthesize nanomaterials and for nanoparticle doping in order to incorporate the red/NIR emission light property into particles that otherwise do not contain the desired emission light. Meso-tetra(4-carboxyphenyl)porphine (TCPP) is one type of porphyrin with a large conjugated π-electron system and four carboxyl groups on its exterior benzene rings. These two key characteristics of TCPP make it ideal for incorporation into GQDs, as it would design and synthesize red-emissive material as well as give rise to excellent water solubility. In this work, TCPP is used in tangent with cis-cyclobutane-1,2-dicarboxylic acid (CBDA-2), a biomass derived organic molecule, to synthesize "green" porphyrin-based graphene quantum dots (PGQDs) with red-emission. The obtained PGQDs were characterized by various analytical methods. Utilizing TEM, HRTEM, and DLS the size distribution of the particles was determined to be 7.9 ± 4.1, well within the quantum dot range of 2-10 nm. FT-IR, XPS, and XRD depicted carbon, nitrogen, and oxygen as the main elemental components with carbon being in the form of graphene and the main porphyrin ring of TCPP remaining present in the final PGQDs product. Lastly, absorption and fluorescence spectroscopy determined the excitation wavelength at 420 nm and the emission at 650 nm which was successfully utilized in the imaging of HeLa cells using confocal microscopy.