Time Resolved Immunofluorometric Assay (TR-IFMA) is a sophisticated bioanalytical technique used in medical research and diagnostics to measure the concentration of specific proteins or molecules present in biological samples. This method is based on the principles of immunoassays, which utilize the specificity of antibodies to detect and quantify analytes of interest.
In TR-IFMA, the measurement is performed with high precision and sensitivity by employing time-resolved fluorescence detection. This technique involves the use of specialized fluorophores, known as lanthanide chelates, as labels on the antibodies. These chelates have unique fluorescence properties that enable the generation of long-lived fluorescence signals, which can be accurately measured after the background fluorescence has decayed.
The assay begins with the addition of the sample, usually serum or plasma, containing the analyte of interest, to a microplate coated with specific antibodies. These antibodies selectively bind to the analyte, forming a complex. After washing away unbound substances, a secondary antibody labeled with the lanthanide chelate is added. This secondary antibody recognizes and binds to a different epitope on the analyte.
To measure the concentration of the analyte, the microplate is then exposed to a specific wavelength of light, causing the lanthanide chelate to emit fluorescence. The emitted fluorescence signal is collected and measured using a time-resolved fluorescence detector, which detects only the long-lived fluorescence emitted by the lanthanide chelate, effectively eliminating any interference from background fluorescence.
By analyzing the intensity of the fluorescence signal, the concentration of the analyte can be determined accurately and with high sensitivity. TR-IFMA is widely used in clinical and research laboratories to detect and quantify proteins, hormones, drugs, and other analytes, playing
