Time Resolved Immunofluorometric Assays (TR-IFMAs) are a type of laboratory technique used in biomedical research and clinical diagnostics to measure and quantify specific biomolecules, such as proteins or antibodies, present in a biological sample.
TR-IFMAs are based on the principles of immunofluorescence and utilize specialized reagents called fluorophores. These fluorophores emit light of different wavelengths when excited by an external energy source, such as a laser. In TR-IFMAs, the fluorophores used are designed to have a long fluorescent lifetime, meaning they continue to emit fluorescent light for an extended period after excitation.
The assay involves first capturing the target biomolecule of interest onto a solid support, such as a microtiter plate or bead. This can be achieved through the specific binding of an antibody or another molecule with affinity for the target. Next, a labeled antibody conjugated with the long-lifetime fluorophore is introduced, which binds to the captured biomolecule forming a complex. After removing any excess unbound components, the sample is then exposed to the excitation source, and the emitted fluorescent light is measured and analyzed.
The advantage of TR-IFMAs over conventional immunofluorescence techniques is their ability to discriminate against background fluorescence, effectively reducing interference from non-specific binding or auto-fluorescence. Additionally, the long fluorescent lifetime of the labels allows for delayed measurements, enabling better temporal resolution and accuracy.
TR-IFMAs find applications in diverse areas of research and clinical practice, such as the detection and quantification of disease-related biomarkers, monitoring of therapeutic drug levels, and studying cellular interactions. They provide researchers and clinicians with a sensitive, precise, and time-resolved method to analyze biomolecular interactions and
