Image scanning microscopy: a vectorial physical optics analysis

Image scanning microscopy (ISM) achieves resolution beyond the diffraction limit by a factor of √2. However, prior ISM research predominantly employs scalar diffraction theory, neglecting critical physical effects such as polarization, aberrations, and Stokes shift. This paper presents a comprehensive vectorial ISM point spread function (PSF)...

Quantifying the minimum localization uncertainty of image scanning localization microscopy

Modulation enhanced single-molecule localization microscopy (meSMLM), where emitters are sparsely activated with sequentially applied patterned illumination, increases the localization precision over single-molecule localization microscopy (SMLM). The precision improvement of modulation enhanced SMLM is derived from retrieving the position of an...

Bayesian posterior density estimation reveals degeneracy in three-dimensional multiple emitter localization

Single-molecule localization microscopy requires sparse activation of emitters to circumvent the diffraction limit. In densely labeled or thick samples, overlap of emitter images is inevitable. Single-molecule localization of these samples results in a biased parameter estimate with a wrong model of the number of emitters. On the other hand,...

Retrieving Pulsatility in Ultrasound Localization Microscopy

Ultrasound localization microscopy (ULM) is a vascular imaging method that provides a 10-fold improvement in resolution compared to ultrasound Doppler imaging. Because typical ULM acquisitions accumulate large numbers of synthetic microbubble (MB) trajectories over hundreds of cardiac cycles, transient hemodynamic variations such as pulsatility...

Precision in iterative modulation enhanced single-molecule localization microscopy

Modulation enhanced single-molecule localization microscopy (meSMLM) methods improve the localization precision by using patterned illumination to encode additional position information. Iterative meSMLM (imeSMLM) methods iteratively generate prior information on emitter positions, used to locally improve the localization precision during...

Theoretical minimum uncertainty of single-molecule localizations using a single-photon avalanche diode array

Single-photon avalanche diode (SPAD) arrays can be used for single-molecule localization microscopy (SMLM) because of their high frame rate and lack of readout noise. SPAD arrays have a binary frame output, which means photon arrivals should be described as a binomial process rather than a Poissonian process. Consequentially, the theoretical...