Single-molecule localization microscopy analysis with ImageJ

ImageJ is a versatile and powerful tool for quantitative image analysis in microscopy. It is open-source software, platform-independent and enables students and researchers to obtain an easy but thorough introduction into image analysis. Especially the image processing package Fiji is a valuable and powerful extension of ImageJ. Several plugins and macros for single-molecule localization microscopy (SMLM) have been developed during the last decade. These novel tools cover the steps from single-molecule localization and image reconstruction to SMLM data postprocessing such as density analysis, image registration or resolution estimation. This article describes how ImageJ/Fiji can be used for image analysis, reviews existing extensions for SMLM, and aims at introducing and motivating novices and advanced SMLM users alike to explore the possibilities of ImageJ/Fiji for automated and quantitative data analysis

DeepCEL0 for 2D Single Molecule Localization in Fluorescence Microscopy

In fluorescence microscopy, Single Molecule Localization Microscopy (SMLM) techniques aim at localizing with high precision high density fluorescent molecules by stochastically activating and imaging small subsets of blinking emitters. Super Resolution (SR) plays an important role in this field since it allows to go beyond the intrinsic light diffraction limit. In this work, we propose a deep learning-based algorithm for precise molecule localization of high density frames acquired by SMLM techniques whose $\ell_{2}$-based loss function is regularized by positivity and $\ell_{0}$-based constraints. The $\ell_{0}$ is relaxed through its Continuous Exact $\ell_{0}$ (CEL0) counterpart. The arising approach, named DeepCEL0, is parameter-free, more flexible, faster and provides more precise molecule localization maps if compared to the other state-of-the-art methods. We validate our approach on both simulated and real fluorescence microscopy data