Why use time-resolved fluorescence detection:
It is known that fluorescent immunoassays using commonly used fluorescein as a marker are often affected by the interference of background fluorescence, such as the interference of the sample carrier, the serum component, and the specular light of the instrument excitation source, so that the sensitivity is greatly limited. Time-resolved fluorescence immunoassay (TR-FIA) is a new detection technique that improves on this shortcoming.
The basic principle of time-resolved fluorescence immunoassay is the use of lanthanide lanthanum (Eu) chelate as a fluorescent marker. The use of such fluorescent substances has the characteristics of long fluorescence lifetime, prolonging the fluorescence measurement time, and the natural background fluorescence to be short-lived. After complete decay, the signal is completely fluorescent for the long-lived lanthanide chelate, thereby effectively eliminating the interference of non-specific background fluorescence.
This technology is used in our HG-1000 time-resolved immunofluorescence analyzer.
Instruments used in time-resolved fluorescence immunoassays:
The time-resolved fluorescence immunoassay used in the time-resolved fluorescence analyzer is different from the general fluorescence spectrophotometer. The pulse light source is mainly used, and the sample is briefly extinguished after the sample is irradiated. The electronic device controls the delay time and waits for the non-specific background fluorescence. After the recession, the long lanthanide fluorescence emitted by the sample is measured. The detection sensitivity can reach 0.2~1ng/ml.
On the reagent: In addition to labeling the reagent antigen or reagent antibody with the lanthanide lanthanum (Eu) chelate, the reagent reacts with the corresponding antibody or antigen in the specimen, and the enhancement solution is used to enhance the fluorescence signal because the immune reaction is completed. The generated antigen-antibody-quinone label complex is weakly generated in a weakly alkaline solution after excitation. In the enhancement solution, the pH can be adjusted to 2 to 3, and the ruthenium ion is easily dissociated, and the β-diketone body in the enhancement liquid generates a new ruthenium chelate with strong fluorescence, which is greatly advantageous for fluorescence measurement.
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