A workflow is a set of components assembled in some specific order to

  1. Measure and estimate some numerical parameters of the biological system or
  2. Visualization

for addressing a biological question. Workflows can be a combination of components from the same or different software packages using several scripts and manual steps.

SpotDetectionIJ

Description

This is a classical workflow for spot detection or blob like structures (vesicules, melanosomes,...)

Step 1 Laplacian of Gaussian to enhance spots . Paraeters= radius, about the average spot radius

Step 2 Detect minima (using Find Maxima with light background option to get minima). Parameter : Tolerance to Noise: to be tested, hard to predict. About the height of the enhanced feautures peaks

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spot detection

Creating an ImageJ plugin / command

Description

The best way to start writing an ImageJ2 plugin (ImageJ2 developers call it command and not plugin) is to download the example command from github and modify it. There is a video tutorial on the whole workflow on how to do this on youtube.

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3-D Density Kernel Estimation

Description

3-D density kernel estimation (DKE-3-D) method, utilises an ensemble of random decision trees for counting objects in 3D images. DKE-3-D avoids the problem of discrete object identification and segmentation, common to many existing 3-D counting techniques, and outperforms other methods when quantification of densely packed and heterogeneous objects is desired. 

Find Maxima (Python)

Description

 

Maxima finding algorithm 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.

find maxima comparison.

Introduction to ImageJ macro language

Biologist
Microscopist

In this session, we will cover the basics of ImageJ macro programming using a simple example: how to quantify signal enrichment at the nuclear rim? Trainees will (re)discover how to record actions, plan a workflow and organise their code. This session will alternate presentation of technical points, to be directly applied during practical exercises. The macro will progressively complexify as new notions are taught.

2D Gaussian fitting macro (Fiji/ImageJ) for multiple signals.

Description

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). 

ZEN Intellesis Trainable Segmentation

Description

Perform Advanced Image Segmentation and Processing across Microscopy Methods
 

Overcome the bottleneck of segmenting your Materials Science images and use ZEISS ZEN Intellesis, a module of the digital imaging software ZEISS ZEN.
Independent of the microscope you used to acquire your image data, the algorithm of ZEN Intellesis will provide you with a model for automated segmentation after training. Reuse the model on the same kind of data and beneft from consistent and repeatable segmentation, not influenced by the operator. 
ZEN Intellesis offers a straightforward, ease-to-use workflow that enables every microscope user to perform advanced segmentation tasks rapidly.

Highlights

  • Simple User Interface for Labelling and Training
  • Integration into ZEN Measurement Framework
  • Support for Multi-dimensional Datasets
  • Use powerful machine learning algorithms for pixel-based classifcation
  • Real Multi-Channel Feature Extraction
  • Engineered Feature Set and Deep Feature Extraction on GPU
  • IP-Function for creating masks an OAD-enabled for advanced automation
  • Powered by ZEN and Python3 using Anaconda Python Distribution
  • Just label objects, train your model and segment your images – there is no need for expert image analysis skills
  • Segment any kind of image data in 2D or 3D. Use data from light, electron, ion or x-ray microscopy, or your mobile phone
  • Speed up your segmentation task by built-in parallelization and GPU (graphics processing unit) acceleration
  • Increase tolerance to low signal-to-noise and artifact-ridden data
  • Seamless integration in ZEN framework and image analysis wizard
  • Data agnostic
  • Compatibility with 2D, 3D and up to 6D datasets
  • Export of multi-channel or labeled images
  • Exchange and sharing of models
  • GPU computing
  • Large data handling
  • Common and well-established machine learning algorithms
  • SW Trial License available

Fit a model for the growth of yeast cells

Description

This notebook uses the rOMERO-gateway and EBImage to process an Image associated to the paper 'Timing of gene expression in a cell-fate decision system'.

The Image "Pos22" is taken from the dataset idr0040-aymoz-singlecell/experimentA/YDA306_AGA1y_PRM1r_Mating. It is a timelapse Image with 42 timepoints separated by 5 minutes. This Image is used to fit a model for the growth of the yeast cells. The notebook does not replicate any of the analysis of the above mentioned paper.

Its purpose is mainly to demonstrate the use of Jupyter, rOMERO-gateway and EBimage.

 

What it does:

  • For each time point of one movie:
    • Read the image for this time point  from the IDR
    • Threshold the images and count the cells using EBimage functions
  • Fit an exponential model to the count of cells against time to get a coefficient of grow (exponential factor)

 

 

 

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Cell or particle counting and scoring the percentage of stained objects

Description

This one example workflow from the Cell Profiler(CP)  Examples . CP is commonly used to count cells or other objects as well as percent-positives, by measuring the per-cell staining intensity. This pipeline shows how to do both of these tasks, and demonstrates how various modules may be used to accomplish the same result. 

In a few words, it used the IdentifyPrimaryObject module of CellProfiler to detect nuclei from a channel (e.g DAPI), then again the same module on another channel to detect another probe (e.g some particular histone)  .

Then objects (nuclei) are related to the second object (Histone), to create a parent child-relation ship: where nuclei can have histone has child. Nuclei are then filtered according to the property of having histone (positive) or not having histone (negtiveobject) related to them.  If needed, nuclei can be expanded in order to include touching object rather than object inside only.

The percentage of positive nuclei vs total number of nuclei can then be computed using the CalculateMath Module.

Positivepercentcell