Automated

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Perfect for the beginner, this demo illustrates simple object detection (segmentation, feature extraction), measurement, and filtering. Requires the Image Processing Toolbox (IPT) because it demonstrates some functions supplied by that toolbox, plus it uses the "coins" demo image supplied with that toolbox. If you have the IPT (you can check by typing ver on the command line), you should be able to run this demo code simply by copying and pasting this code into a new editor window, and then clicking the green "run" triangle on the toolbar. First finds all the objects, then filters results to pick out objects of certain sizes. The basic concepts of thresholding, labeling, and regionprops are demonstrated with a simple example.

It's a good tutorial for those users new to MATLAB's image processing capabilities to learn on, before they go on to more sophisticated algorithms.

idTracker is a videotracking software that keeps the correct identity of each individual during the whole video. It works for many animal species including mice, insects (Drosophila, ants) and fish (zebrafish, medaka, stickleback). idTracker distinguishes animals even when humans cannot, such as for size-matched siblings, and reidentifies animals after they temporarily disappear from view or across different videos. It is robust, easy to use and general. Technique details and analyses of several applications are described in Pérez-Escudero et al (2014).

Video protocol: https://www.youtube.com/watch?v=oC9tp5TKAyw

Example image: Example video of 5 zebrafish

Using a text file containing 3D point coordinates as reference pairs, 3D image stack is transformed.

DBSCAN (Density-based spatial clustering of applications with noise) performs multi-dimensional clustering based on the local density of points. This plugin is implemented for 2-3 dimensions.

Two workflows are proposed here:

one based on fiducials, the other one on cross-correlation.

A clear tutorial on how to write a MATLAB script to segment clustered cells.

The full script is downloadable near the bottom of the article. 

Matlab toolbox to analyze single molecule mRNA FISH data. Allows counting the number of mature and nascent transcripts in 3D images. See 2513. Following toolboxes are required: - Optimization toolbox - Statistics toolbox - Image processing toolbox - (Optional) Parallel processing toolbox

Input data type: 3D image

Output data type: CSV

A Jython script using the plugin : Register Virtual Stack Slices It takes a sequence of image slices stored in a folder, and delivers a list of registered image slices (with enlarged canvas). One of the images in the sequence can be selected by the user as reference and it will remain intact. The plugin can perform 6 types of image registration techniques: - Translation - Rigid (translation + rotation) - Similarity (translation + rotation + isotropic scaling) - Affine - Elastic (via bUnwarpJ with cubic B-splines) - Moving least squares All models are aided by automatically extracted SIFT features.

The rapidSTORM project is an open source evaluation tool that provides fast and highly configurable data processing for single-molecule localization microscopy such as dSTORM. It provides both two-dimensional and three-dimensional, multi-color data analysis as well as a wide range of filtering and image generation capabilities. The general operation of rapidSTORM is described in Wolter et al (2012).

(from the webpage) >This usage example shows how to produce two-color images from spectrally unmixed data sets. It was written for an Alexa647/Alexa700 measurement on the Würzburg 1 biplane setup as documented in [Aufmkolk2012]. The first two tasks in this example produce prerequisite knowledge for the image generation, the alignment information (Produce linear alignment matrix) and the F2 ratios, i.e. the relative intensity of fluorophores between the channels. [Aufmkolk2012] Hochauflösende Mehrfarben-Fluoeszenzmikroskopie. Sarah Aufmkolk. Julius-Maximilians-Universität Würzburg. 2012-mar.

The ImageJ pligin, called PixFRET, allows a simple and rapid determination of channel bleed-through parameters and the display of normalized FRET images. see 2521

Input data type: 

Stacks with 2 channels (controls for bleed-through calculation) or 3 channels (FRET calculation)

Output data type: 

One Image with FRET values and One image with normalized FRET values

This macro is a plugin macro to the "Intelligent Imaging" workflow. It detects the Cytoo patterns (specific fluorsecence channel) and computes the occupancy (number of cells) of each pattern by analyzing the images of the DAPI channel. The analysis function can be easily extended to, for instance, only select the cells that are well spread on the patterns (by analyzing a third channel with a properly chosen marker of the cytoplasm).

as of 20180529, links are not working due to web defacement.

The Measure Rosette Area Tool allows to measure the area of the rosettes of arabidopsis plants.

See: http://dev.mri.cnrs.fr/projects/imagej-macros/wiki/Measure_Rosette_Area…

Example data: http://biii.eu/node/1146, http://biii.eu/node/1145

An example macro introduced in the documentation page of the ImageJ plugin Trainable Weka Segmentation (in Fiji, it's bundled). A segmentation protocol based on machine learning. Full macro is available in the "Download" Link. 

This plugin can be trained to learn from the user input and perform later the same task in unknown (test) data. Weka: it makes use of all the powerful tools and classifiers from the latest version of Weka. Segmentation: it provides a labeled result based on the training of a chosen classifier. Trainable Weka Segmentation Complete macro example is at the end of the page.

A clear tutorial for splitting connected particles (cells) in a binary mask.

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Fluorescence in situ hybridization (FISH) is used to study the organization and the positioning of specific DNA sequences within the cell nucleus. Analyzing the data from FISH images is a tedious process that invokes an element of subjectivity. Automated FISH image analysis offers savings in time as well as gaining the benefit of objective data analysis. While several FISH image analysis software tools have been developed, they often use a threshold-based segmentation algorithm for nucleus extraction. As fluorescence signal intensities can vary significantly from experiment to experiment, from cell to cell, and within a cell, threshold based segmentation is inflexible and often insufficient for automatic image analysis, leading to additional manual extraction and potential subjective bias. To overcome these problems, we developed a graphical software tool called FISH Finder to automatically analyze FISH images that vary significantly. By posing the nucleus extraction as a classification problem, compound Bayesian Classifier is employed so that contextual information is utilized, resulting in reliable classification and boundary extraction. This makes it possible to analyze FISH images efficiently and objectively without adjustment of input parameters.

This plugin will return on a full 256-grey level image (limitation in this version) or on a ROI several texture features such as described in Haralick publication. Can be run in batch mode.

This Javascript works in ImageJ to measure 3D intensity profile along cylindrical space with variable radius.

The website implements a set of computer vision algorithms designed to automatically process time-lapse images of fluorescently labeled focal adhesion proteins in motile cells.

The methods associated with the processing have been published in PLOS One and Cell. The publication describes a quantitative analysis of focal adhesion dynamics that have been imaged using TIRF. All image processing steps are well explained or referenced.

To better understand the dynamic regulation of focal adhesions, we have developed an analysis system for the automated detection, tracking, and data extraction of these structures in living cells. This analysis system was used to quantify the dynamics of fluorescently tagged Paxillin and FAK in NIH 3T3 fibroblasts followed via Total Internal Reflection Fluorescence Microscopy (TIRF). High content time series included the size, shape, intensity, and position of every adhesion present in a living cell. These properties were followed over time, revealing adhesion lifetime and turnover rates, and segregation of properties into distinct zones.

The workflow computes cell-based colocalisation of two stainings in 2-D images. Both pixel- and object-based readouts are provided and some pros and cons are discussed. Please read here for more information:

https://github.com/tischi/ImageAnalysisWorkflows/blob/master/CellProfil…

Input data type: 

images

Output data type: 

processed images, numbers, text file, csv files