Untersuchung von alternativen Lichtquellen-Konzepten für die HTRF®-Messung des Mikrotiterplatten-Readers Infinite® M1000

The aim of this thesis is the improvement of the homogeneous time resolved fluorescence (HTRF®) performance of the Infinite® M1000 microplate reader with monochromators from the company Tecan. The main focus was lain on increasing the signal strength of the measured HTRF® assay, by sending more light energy to the assay sample. In doing so, the current performance of the excitation side, consisting of a flash lamp coupled to a double monochromator, should be improved by using a different light source concept. Therefore the improvement of quality, the effort and the integration of the excitation source into the reader, as well as the measurement set-up are the important factors considered in this work. To this end a UV-LED and Xe-flash lamp solutions with a filter instead of the double monochromator were investigated.

After familiarization with the functions of the Infinite® M1000 reader, the HTRF® theory and the measured HTRF® assay, the measurement system and the parameters to be optimized had to be defined. The design comprised also the necessary electronics and firmware as well as the required optic elements which had to be designed.

During the project, other options which could lead to an improvement of the HTRF® performance of the Infinite® M1000 reader also had to be considered and investigated.

The results of the UV-LED measurements do not show a clear improvement, as the light power of UV-LEDs are still too low. The measurements conducted with the flash lamp modules and especially with the flash lamp (40 W), show improvement of the variance of the measurement as well as the detected signal strength of the HTRF® measurement.

In the course of this work several possibilities were tested and some of them lead to improvements in the measurement method, which will influence the future developments of the device.