Addressing systematic errors in axial distance measurements in single-emitter localization microscopy

Minimizing detection errors in single molecule localization microscopy.

Fluorescence microscopy using single molecule imaging and localization (PALM, STORM, and similar approaches) has quickly been adopted as a convenient method for obtaining multicolor, 3D superresolution images of biological samples. Using an approach based on extensive Monte Carlo simulations, we examined the performance of various noise reducing filters required for the detection of candidate m...

Subresolution axial distance measurements in far-field fluorescence microscopy with precision of 1 nanometer

Sub-resolution Axial Localization of Nanoparticles in Fluorescence Microscopy

In recent years single particle tracking has significantly contributed to the study of intracellular molecular dynamics [2]. Particle tracking primarily consists of localizing particles in three dimensions (3-D) from a number of images acquired by fluorescence microscopy. While many approaches have been proposed for the lateral (i.e., x-y) localization of particles from acquisitions that are (a...

Challenges in quantitative single molecule localization microscopy.

Single molecule localization microscopy (SMLM), which can provide up to an order of magnitude improvement in spatial resolution over conventional fluorescence microscopy, has the potential to be a highly useful tool for quantitative biological experiments. It has already been used for this purpose in varied fields in biology, ranging from molecular biology to neuroscience. In this review articl...

Localization accuracy in single-molecule microscopy.

One of the most basic questions in single-molecule microscopy concerns the accuracy with which the location of a single molecule can be determined. Using the Fisher information matrix it is shown that the limit of the localization accuracy for a single molecule is given by, lambda(em)/2pi n(a) square root of gammaAt, where lambda(em), n(a), gamma, A, and t denote the emission wavelength of the ...