Simultaneous detection of 3D orientation and 3D spatial localization of single fluorescent emitters

Oumeng Zhang, Matthew D. Lew

Precisely measuring the three-dimensional position and orientation of individual fluorophores is challenging due to the substantial photon shot noise in single-molecule experiments. Facing this limited photon budget, numerous techniques have been developed to encode 2D and 3D position and 2D and 3D orientation information into fluorescence images. In this work, we adapt classical and quantum estimation theory and propose a mathematical framework to derive the best possible precision for measuring the position and orientation of dipole-like emitters for any fixed imaging system. We find that it is impossible to design an instrument that achieves the maximum sensitivity limit for measuring all possible rotational motions. Further, our vectorial dipole imaging model shows that the best quantum-limited localization precision is 4%-8% worse than that suggested by a scalar monopole model. Overall, we conclude that no single instrument can be optimized for maximum precision across all possible 2D and 3D localization and orientation measurement tasks.

Pradipta Panchadhyayee, Bibhas Kumar Dutta, Nityananda Das +1 more

A. D�barre, Rodolphe Jaffiol, Carine Julien +5 more

Luca Maggio, Vincenzo Tamma

Abstract We present a quantum sensing protocol for the simultaneous estimation of the difference in the localization parameters of two single-photon sources, paving the way to single-photon 3D imaging and 3D nanoscopy beyond the diffraction limit. This is achieved by exploiting two-photon interference of the two emitted photons at a beam splitter via sampling measurements in the frequency and transverse momenta at the output. We prove theoretically that this technique reaches the ultimate sensitivity in the 3D relative localization of two emitters, already with a number of sampling measurements of $$\sim 2000$$ ∼ 2000 and a bias in the three localization parameters below $$1\%$$ 1 % . These results are independent of the values of the localization parameters to estimate.

Wolfgang Alt

Dan Oron, Ron Tenne, Yonatan Israel +1 more

Far-field optical microscopy beyond the Abbe diffraction limit, making use of nonlinear excitation (e.g. STED), or temporal fluctuations in fluorescence (PALM, STORM, SOFI) is already a reality. In contrast, overcoming the diffraction limit using non-classical properties of light is very difficult to achieve due to the fragility of quantum states of light. Here, we experimentally demonstrate superresolution microscopy based on quantum properties of light naturally emitted by fluorophores used as markers in fluorescence microscopy. Our approach is based on photon antibunching, the tendency of fluorophores to emit photons one by one rather than in bursts. Although a distinctively quantum phenomenon, antibunching is readily observed in most common fluorophores even at room temperature. This nonclassical resource can be utilized directly to enhance the imaging resolution, since the non-classical far-field intensity correlations induced by antibunching carry high spatial frequency information on the spatial distribution of emitters. Detecting photon statistics simultaneously in the entire field of view, we were able to detect non-classical correlations of the second and third order, and reconstructed images with resolution significantly beyond the diffraction limit. Alternatively, we demonstrate the utilization of antibunching for augmenting the capabilities of localization-based superresolution imaging in the presence of multiple emitters, using a novel detector comprised of an array of single photon detectors connected to a densely packed fiber bundle. These features allow us to enhance the spatial and temporal resolution with which multiple emitters can be imaged compared with other techniques that rely on CCD cameras.