Image analysis

Fiji plugin to segment oocyte and zona pellucida contours from transmitted light images and extract hundreds of morphological features to describe numerically the oocyte. Segmentation is based on trained neural networks (U-Net) that were trained on both mouse and human oocytes (in prophase and meiosis I) acquired in different conditions. They are freely avaialable on the github repository and can be retrained if necessary. Oocytor also have options to extract hundreds of morphological/intensity features to characterize manually the oocyte (eg perimeter, texture...). These features can also be used in machine learning pipeline for automatic phenotyping.

The BioVoxxel Toolbox is a suite which contains plugins and some macros dealing with image filtering, image segmentation and binary image processing and analysis. The following plugins are hosted here:

• Extended Particle Analyzer
• Binary Feature Extractor
• Speckle Inspector
• Watershed Irregular Features
• EDM Binary Operations
• Filter Check
• Pseudo flat-field correction
• Convoluted Background Subtraction
• 2D Particle Distribution (Distribution_Analysis)
• Cluster Indicator
• SSIDC Cluster Indicator
• Gaussian weighted Median filter
• Adaptive Filter
• Enhance True Color Contrast
• Mode and Differential Limited Mean Binarization
• Basic Recursive Filter

MATLAB app to characterize nanoparticles imaged with super-resolution microscopy. nanoFeatures will read text and csv files from the NIKON and ONI microscopes and from the ThunderSTORM Fiji plugin, then cluster the localizations and filter by size and sphericity and finally output nanoparticle features like size, aspect ratio, and number of localizations per cluster (total and for each channel).

Fractal is a framework to process high-content imaging data at scale and prepare it for interactive visualization. Fractal provides distributed workflows that convert TBs of image data into OME-Zarr files. The platform then processes the 3D image data by applying tasks like illumination correction, maximum intensity projection, 3D segmentation using cellpose and measurements using napari workflows. The pyramidal OME-Zarr files enable interactive visualization in the napari viewer.

MiNA is a simplified workflow for analyzing mitochondrial morphology using fluorescence images or 3D stacks in Fiji. The workflow makes use of ImageJ Ops, 3D Viewer, Skeletonize (2D/3D), Analyze Skeleton, and Ridge Detection. In short, the tool estimates mitochondrial footprint (or volume) from a binarized copy of the image as well as the lengths of mitochondrial structures using a topological skeleton. The values are reported in a table and overlays (or a 3D rendering) are generated to assess the accuracy of the analysis.

Relate is a correlative software package optimised to work with EM, EDS, EBSD, & AFM data and images.  It provides the tools you need to correlate data from different microscopes, visualise multi-layered data in 2D and 3D, and conduct correlative analyses.

• Combining data from different imaging modalities (e.g. AFM, EDS & EBSD)

• Interactive display of multi-layer correlated data

• Analytical tools for metadata interrogation

• Documented workflows and processes

Correlate

• Import data from AZtec using the H5oina file format
• Import AFM data
• Correlate both sets of data using intuitive image overlays and image matching tools
• Produce combined multimodal datasets

Visualise

• 2D display of multi-layered data
• 3D visualisation of topography combined with AFM material properties, EM images, and EDS & EBSD map overlays
• Customisation of colour palettes, data overlays, image rendering options, and document display
• Export images and animations

Analyse

• Generate profile (cross section) views of multimodal data
• Measure and quantify data across multiple layers
• Analyse areas via data thresholding using amount of x-ray counts, phase maps, height, or other material properties.
• Select an extensive range of measurement parameters
• Export analytical data to text or CSV files

ModularImageAnalysis (MIA) is an ImageJ plugin which provides a modular framework for assembling image and object analysis workflows. Detected objects can be transformed, filtered, measured and related. Analysis workflows are batch-enabled by default, allowing easy processing of high-content datasets.

MIA is designed for “out-of-the-box” compatibility with spatially-calibrated 5D images, yielding measurements in both pixel and physical units.  Functionality can be extended both internally, via integration with SciJava’s scripting interface, and externally, with Java modules that extend the MIA framework. Both have full access to all objects and images in the analysis workspace.

Workflows are, by default, compatible with batch processing multiple files within a single folder. Thanks to Bio-Formats, MIA has native support for multi-series image formats such as Leica .lif and Nikon .nd2.

Workflows can be automated from initial image loading through processing, object detection, measurement extraction, visualisation, and data exporting. MIA includes near 200 modules integrated with key ImageJ plugins such as Bio-Formats, TrackMate and Weka Trainable Segmentation.

Module(s) can be turned on/off dynamically in response to factors such as availability of images and objects, user inputs and measurement-based filters. Switches can also be added to “processing view” for easy workflow control.

MIA is developed in the Wolfson Bioimaging Facility at the University of Bristol.

ClearMap is a toolbox for the analysis and registration of volumetric data from cleared tissues.

It was initially developed to map brain activity at cellular resolution in whole mouse brains using immediate early gene expression. It has since then been extended as a tool for the qunatification of whole mouse brain vascualtur networks at capilary resolution.

It is composed of sevral specialized modules or scripts: tubemap, cellmap, WobblyStitcher.

ClearMap has been designed to analyze O(TB) 3d datasets obtained via light sheet microscopy from iDISCO+ cleared tissue samples immunolabeled for proteins. The ClearMap tools may also be useful for data obtained with other types of microscopes, types of markers, clearing techniques, as well as other species, organs, or samples.

The napari-pyclesperanto-assistant is a yet experimental napari plugin for building GPU-accelerated image processing workflows targeting life-sciences and bio-image analysis. It is part of the clEsperanto project. It uses pyclesperanto and pyopencl as backend for processing images.

Phindr3D is a comprehensive shallow-learning framework for automated quantitative phenotyping of three-dimensional (3D) high content screening image data using unsupervised data-driven voxel-based feature learning, which enables computationally facile classification, clustering and data visualization.

Please see our GitHub page and the original publication for details.

This tool allows to analyze morphological characteristics of complex roots. While for young roots the root system architecture can be analyzed automatically, this is often not possible for more developed roots. The tool is inspired by the Sholl analysis used in neuronal studies. The tool creates a binary mask and the Euclidean Distance Transform from the input image. It then allows to draw concentric circles around a base point and to extract measures on or within the circles. Instead of circles, which present the distance from the base point, horizontal lines can be used, which present the distance in the soil from the base-line. The following features are currently implemented:

• The area of the root per distance/depth.
• The number of border pixel per distance/depth, giving an idea of the surface in contact with the soil.
• The maximum radius per distance/depth of a root, measured at the crossing points with the circles or lines.
• The number of crossings of roots with the circles or lines.
• The maximum distance to the left and the right from the vertical axis at crossing points with the circles or lines.

Quote:

LaRoME = Label + Region Of Interest + Measure

Label image (aka Count Masks): An image in which pixels of an object have all the same value. Each object has a unique value.

Measurement image: An image in which pixels of an object have all the same value, corresponding to a measurement (Area, Angle, Mean...)

The ImageM application proposes an integrated user interface that facilitates the processing and the analysis of multi-dimensional images within the Matlab environment. It provides a user-friendly visualization of multi-dimensional images, a collection of image processing algorithms and methods for analysis of images, the management of spatial calibration, and facilities for the analysis of multi-variate images. Its graphical user interface is largely inspired from the open source software "ImageJ". ImageM can also be run on the open source alternative software to Matlab, Octave.

ImageM is freely distributed on GitHub: https://github.com/mattools/ImageM.

Epina ImageLab is a Microsoft Windows-based multisensor imaging tool for processing and analyzing hyperspectral images. It is a modular system consisting of a basic engine, a graphical user interface, a chemometrics toolbox and optional user-supplied modules. It supports the most important spectroscopic imaging techniques, such as UV/Vis, infrared, Raman, THz, optical emission/absorption, and mass spectrometry. On top of that Epina ImageLab enables the user to merge hyperspectral images with maps of physical properties and conventional high-resolution color photos. 

QuickFit 3 is a data evaluation software for FCS Fluorescence Correlation Spectroscopy and imagingFCS (imFCS) measurements, developed in the group B040 (Prof. Jörg Langowski) at the German Cancer Research Center (DKFZ). Actually QuickFit 3 itself is a project manager and all functionality is added as plugins. A set of tested plugins for FCS, imagingFCS and some microscopy-related image processing tasks is supplied together with the software.

OligoMacro Toolset, is an ImageJ macro-toolset aimed at isolating oligodendrocytes from wide-field images, tracking isolated cells, characterizing processes morphology along time, outputting numerical data and plotting them. It takes benefit of ImageJ built-in functions to process images and extract data, and relies on the R software in order to generate graphs.

Protein array is used to analyze protein expressions by screening simultaneously several protein-molecule interactions such as protein-protein and protein-DNA interactions. In most cases, the detection of interactions leads to an image containing numerous lines of spots that will be analyzed by comparing tables of intensity values. To describe the observed different patterns of expression, users generally show histograms with the original associated images [1]. The “Protein Array Analyzer” gives a friendly way to exploit this type of analysis, thus allowing quantification, image modeling and comparative analysis of patterns.

The Protein Array Analyzer, which was programmed in ImageJ’s macro language, is an extention of the Dot Blot Analyzer, [2], [3] a graphically interfaced tool that greatly simplifying analysis of dot arrays.

Fiji plugin for detecting, tracking and quantifying filopodia

CellProfiler Analyst (CPA) allows interactive exploration and analysis of data, particularly from high-throughput, image-based experiments. Included is a supervised machine learning system which can be trained to recognize complicated and subtle phenotypes, for automatic scoring of millions of cells. CPA provides tools for exploring and analyzing multidimensional data, particularly data from high-throughput, image-based experiments analyzed by its companion image analysis software, CellProfiler.

The Binary Pattern Dictionary Learning (BPDL) package is suitable for image analysis on a set/sequence of images to determine an atlas of a compact region. In particular, the application can be maping gene activation accross many samples, brain activations in a time domain, etc.

ROI measurement plug-in for Icy.

Set of Tools for super resolution microscopy

This workflow describes a semi-automatic image segmentation procedure for 3D reconstructions of the coronary arterial tree, after which how different morphometric features are automatically extracted, including vessel lumen diameter of the three main coronaries.

The macro generates orthogonal projections from bead images along the lateral and axial dimensions which are displayed using a customized look-up-table to color code intensities. A Gaussian curve is fit to the intensity profile of a fluorescent bead image and full-with-at-half-maximum (FWHM) values are extracted, and listed next to theoretical values for comparison. 

NanoJ-SQUIRREL (Super-resolution Quantitative Image Rating and Reporting of Error Locations) is a software package designed for assessing and mapping errors and artefacts within super-resolution images. This is achieved through quantitative comparison with a reference image of the same structure (typically a widefield, TIRF or confocal image). SQUIRREL produces quantitative maps of image quality and resolution as well as global image quality metrics.

InspectJ is a free ImageJ/FIJI tool to inspect digital image integrity.

InspectJ_v2 is a newer version for advanced users. It applies additional features like histogram equalization and gamma correction for improved image inspections.

AutoPilot is the open source project that hosts the general algorithm for fast and robust assessment of local image quality, an automated computational method for image-based mapping of the three-dimensional light-sheet geometry inside a fluorescently labeled biological specimen, and a general algorithm for data-driven optimization of the system state of light-sheet microscopes capable of multi-color imaging with multiple illumination and detection arms.

Maxima finding algorithm implemented in Python recreated from implementation in Fiji(ImageJ)

This is a re-implementation of the java plugin written by Michael Schmid and Wayne Rasband for ImageJ. The original java code source can be found in: https://imagej.nih.gov/ij/developer/source/ij/plugin/filter/MaximumFinder.java.html 

This implementation remains faithful to the original implementation but is not 100% optimised. The java version is faster but this could be alleviated by compiling c code for parts of the code. This script is simply to provide the functionality of the ImageJ find maxima algorithm to individuals writing pure python script.

The algorithm works as follows:

The first stage in the maxima finding algorithm is to find the local maxima. This involves processing the image with a 3x3 neighbourhood maximum filter. Once filtered this image is compared back to the original, where the pixels are the same value represents the locations of the local maxima. Typically there are far too many local maxima to be meaningful so the goal is then to merge and prune this maxima using some kind of measure of quality. In the case of algorithm a single parameter is used, the noise tolerance (Prominence). If a maxima is close to another then the maxima will be merged or removed based on the below criteria.

Starting with the brightest maxima and working down the intensities:

• Expand out (‘flood fill’) from each maxima location. Neighbouring pixels within a noise tolerance (notl) of the maxima are scanned until the region within tolerance is exhausted.
  • If the pixels are equal to the maxima, mark this as equal.
  • If a greater maxima is met, ignore the active maxima.
  • If the pixels are less than maxima, but greater than maxima minus the noise tolerance, mark as listed.
  • Mark all ‘listed’ pixels 'processed' if they are included within a valid peak region, otherwise reset them.
  • From the regions containing a peak, calculate the best pixel to be considered as maxima based on minimum distance calculation with all those maxima considered equal.
     

For a video detailing how this algorithm works please see:

https://youtu.be/f9vXOMKOlaY

Or for examples of it being used in practise, please see:

https://youtu.be/9wvPsEzRWzI

This script includes a rough feature detection and then fine 2D Gaussian algorithm to fit Gaussians within detected regions. This macro is unique because the ImageJ/Fiji curve fitting API only supports 1-D curve. I get around this by linearising the equation. This implementation is for isotropic (spherical) or anistropic (longer in x/y) diagonally covariant Gaussians but not fully covariant Gaussians (anisotropic and rotated). 

SimpleITK provides a simplified interface to ITK in a variety of languages. A user can either download pre-built binaries, if they are available for the desired platform and language, or SimpleITK can be built from the source code. Currently, Python binaries are available on Microsoft Windows, GNU Linux and Mac OS X. C# and Java binaries are available for Windows. We are also working towards supporting R packaging.

Bisque (Bio-Image Semantic Query User Environment) : Store, visualize, organize and analyze images in the cloud. It also allow to run workflows using a set of deployed tools, such as CellProfiler, RootTipMultin Nuclear Tracker, Microtubule tracker etc...

Bisque was developed for the exchange and exploration of biological images.

The Bisque system supports several areas useful for imaging researchers from image capture to image analsysis and querying. The bisque system is centered around a database of images and metadata. Search and comparison of datasets by image data and content is supported. Novel semantic analyses are integrated into the system allowing high level semantic queries and comparison of image content.

• Bisque is free and open-source
• Flexible textual and graphical annotations
• Cloud scalability: PBs of images, millions of annotations
• Distributed storage: local, iRODS, S3
• Integrated image analysis, high-throughput with Condor
• Analysis in MATLAB, Python, Java+ImageJ
• 100+ biological image formats
• Very large 5D images (100+ GB)

CaPTk is a software platform for analysis of radiographic cancer images, currently focusing on brain, breast, and lung cancer. CaPTk integrates advanced, validated tools performing various aspects of medical image analysis, that have been developed in the context of active clinical research studies and collaborations toward addressing real clinical needs. With emphasis given in its use as a very lightweight and efficient viewer, and with no prerequisites for substantial computational background, CaPTk aims to facilitate the swift translation of advanced computational algorithms into routine clinical quantification, analysis, decision making, and reporting workflow. Its long-term goal is providing widely used technology that leverages the value of advanced imaging analytics in cancer prediction, diagnosis and prognosis, as well as in better understanding the biological mechanisms of cancer development.

Image analysis tools to be used within Galaxy

HyphaTracker propose a workflow for time-resolved analysis of conidia germination. Each part of this workflow can also be used independnatly , as a toolbox. It has been tested on bright-field microscopic images of conidial germination. Its purpose is mainly to identify the germlings and to remove crossing hyphae, and measure the dynamics of their growth.

Kymograph generation under ImageJ:

one simple solution, plot a line (ROI line) on the first frame, where you want to generate the kymograph.

Use

Image  / Stacks  / Reslice

It will generate a new image were Y dimension is the time, and X the position on the line you have drawn.

The MIPAV (Medical Image Processing, Analysis, and Visualization) application enables quantitative analysis and visualization of medical images of numerous modalities such as PET, MRI, CT, or microscopy. Using MIPAV's standard user-interface and analysis tools, researchers at remote sites (via the internet) can easily share research data and analyses, thereby enhancing their ability to research, diagnose, monitor, and treat medical disorders.

A software toolkit for computational morphometry of biomedical images, CMTK comprises a set of command line tools and a back-end general-purpose library for processing and I/O.

The command line tools primarily provide the following functionality: registration (affine and nonrigid; single and multi-channel; pairwise and groupwise), image correction (MR bias field estimation; interleaved image artifact correction; EPI unwarping), processing (filters; combination of segmentations via voting and STAPLE; shape-based averaging), statistics (t-tests; general linear model).

Software for analysis, visualization, simulation, and acquisition  of data from spectroscopy and fluorescence microscopy.

• Fluorescence Correlation Spectroscopy (FCS)
• Fluorescence Lifetime Imaging (FLIM) and Phasor plots
• Förster Resonance Energy Transfer (FRET)
• Generalized Polarization (GP) and Spectral Phasors
• Number and Brightness (N&B)
• Photon Counting Histogram (PCH)
• Raster and Spatio-temporal Image Correlation Spectroscopy (RICS and STICS)
• Single Particle and Modulation Tracking (SPT, MT)
• Image Mean Square Displacement (iMSD)
• Pair correlation function (pCF)