Multi-emitter colocalization in 3D stochastic optical reconstruction microscopy

In this paper, we analyze the Cramer-Rao lower bound on the accuracy of 3-D emitter locations estimated from a STORM movie with a cylindrical lens. Numerical evaluation for randomly and uniformly distributed emitters indicates that as the emitter density in the image plane increases, the Cramer-Rao lower bounds on the average full-width half-maximum (FWHM) of location estimates in the lateral plane and axial direction increase exponentially fast; and the exponents can be numerically accurately estimated. The Cramer-Rao lower bounds are inversely proportional to the square root of mean number of photons of an emitter and monotonically decrease as the signal to noise ratio increases. The result reveals the insightful property of 3-D STORM movies and provides a benchmark for the achievable accuracy of location estimation algorithms. The developed algorithm with multiemitter colocalization can improve the temporal resolution by five folds compared with the single-emitter location estimator.