Single-molecule orientation localization microscopy I: fundamental limits

Abstract

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.

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Cited by in corpus (7)

• Single-molecule orientation localization microscopy II: a performance comparison
• Efficient superoscillation measurement for incoherent optical imaging
• Resolving the Orientations of and Angular Separation between a Pair of Dipole Emitters
• Differentiable optimization of the Debye-Wolf integral for light shaping and adaptive optics in two-photon microscopy
• Dipole-Spread Function Engineering for 6D Super-Resolution Microscopy
• Quantum limits to resolution and discrimination of spontaneous emission lifetimes
• Quantifying classical and quantum bounds for resolving closely spaced, non-interacting, simultaneously emitting dipole sources in optical microscopy