The Brilliant Violet™ family of fluorescent molecules are organic polymers with an extraordinary capacity to absorb energy (extinction coefficient) and a high efficiency with which to convert that absorbed energy to an emitted signal (quantum yield). When conjugated to antibodies, this results in high intensity brightness on labeled cells.
Improvements over Traditional Organic Dyes
Traditional organic dyes are quite small, ranging between 300 - 1200 Da. Although multiple dyes can be conjugated to a single antibody, each acts independently from the others around them, thus their potency is limited to their own structural constraints and the potential for self quenching. In contrast, the Brilliant Violet™ polymers are comparable in size to AmCyan or APC, but unlike a protein, consist of repeating fluorescent subunits that act cooperatively along the entire length of the polymer backbone. Energy is conducted much like a molecular antennae to capture light and pass it down the antennae like a lightning rod. As the subunits work cooperatively, the polymers' extinction coefficient is substantially larger than that of standard organic dyes and this is reflected in superior brightness and signal-to-noise.
Brilliant Violet™ Chemistry
Structurally, Brilliant Violet™ polymers are unsaturated organic materials comprised of alternating single and double bonds and aromatic units. It is this repeating bond structure that creates a continuous π-orbital system and extended electronic delocalization. Such features give rise to unique and tunable optical properties, including large extinction coefficients (>106 M-1cm-1), intense photoluminescence, and massive collective response, all of which help to address fundamental limitations in detection sensitivity. Adaptation of polymer side chain chemistry to impart solubility in aqueous solution confers ability for use as a highly sensitive fluorescent conjugate in biological applications such as flow cytometry and microscopy.
Physical properties, such as high quantum yield in typical flow buffers, high solubility, and minimal non-specfic binding, are all built into the backbone structure and in the side chain modifications. Additionally, the polymer design specifically incorporates well defined functional sites for covalent attachment to antibodies.