| 摘要 |
Time-resolved single molecule fluorescence measurements may be used to probe the conformational dynamics of biological macromolecules. The best time resolution will only be achieved by measuring the arrival times of individual photons at the detector. A method is described for the estimation of molecular parameters based on individual photon arrival times. The amount of information present in a data set is quantified by the Fisher information, thereby providing a guide to deriving the basic equations relating measurement uncertainties and time resolution. Based on these information-theoretical considerations, the present invention presents a data analysis algorithm that details the optimal analysis of single-molecule data. This method natively accounts and corrects for background photons and cross-talk, and can scale to an arbitrary number of channels. By construction, and with corroboration from computer simulations, it is clear that this algorithm reaches the theoretical limit, extracting the maximal information out of the data. |
