Quantitative tracing of bioprobes by simultaneously monitoring radiative and nonradiative relaxations

Abstract

Bioprobe based on fluorescence is widely used in biological and medical research due to its high sensitivity and selectivity. Yet, its quantification in vivo is complicated and often compromised by the interaction between the fluorophore with the environmental factors, as well as the optical scattering and absorption by the tissue. A high florescence quantum yield and minimal interference by the environment are key requirements for designing an effective bioprobe, and the pre-requisitions severely limit the available options. We propose that a comprehensive evaluation of potential bioprobe can be achieved by simultaneously measuring both radiative and non-radiative transitions, the two fundamental and complementary pathways for the energy de-excitation. This approach will not only improve the accuracy of the quantification by catching the information from a broader spectrum of the energy, but also provide additional information of the probe environment that often impacts the balance between the two forms of the energy transition. This work first analyzes the underlying mechanism of the hypothesis. The practical feasibility is then tested by means of simultaneous measurements of photoacoustic signal for the non-radiative and fluorescence for the radiative energy processes, respectively. It is demonstrated that the systematic evaluation of the probe energy de-excitation results in an improved quantitative tracing of a bioprobe in complex environment.