rOMERO-gateway

Description

R wrapper around the OMERO Java Gateway, to enable access to OMERO via R using rJava

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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

Quantification of outer ring diameters of centriole or PCM proteins of cycling HeLa cells in interphase

Description

This workflow can be ran with data from 3D-SIM showing the centrosomes in order to compare the distribution of diameters of rings (or toroids) of different proteins from the centrioles or the peri centriolar material. It aims to reproduce the results of the Nature Cell Biology Paper Subdiffraction imaging of centrosomes reveals higher-order organizational features of pericentriolar material  from the same data set but with a different analysis method.

It is slightly different from the methods described in the paper itself, where the method was to work on a maximum intensity projection of a 3D-SIM stack, and then to fit circle to the centrioles to estimate the diameters of the toroids.

In this workflow, the images are read from the IDR , then process by thresholding (Maximum entropy auto thresholding with Image J), and processed by Analyze Particles  with different measurement sets, including the bouding box. Then the analysis of diameters and the statistical test are performed using R. All the code and data sets are available, and in the case of this paper have shown a layered organisation of the proteins.

Combined view from Figure 1 Lawo et al.

Detection of Molecules - DoM

Description

A collection of components for super resolution image data:

  • Detect Molecules
  • Reconstruct Image
  • Results table
  • Drift correction
  • Chromatic correction

Temporal Medial Filter

Description

This component can be used to find moving foreground features, which can be a powerful way to suppress false background detections in subsequent tracking steps.

set time window, and standard deviations above background for foreground time window should be more than 2x larger than time taken for a feature to traverse a pixel (NB. total window is 2x half-width +1) moving foreground identified by intensity increase relative to background average (i.e. median) for a pixel over a given time window "soft" segmentation, yielding foreground probability related to excess intensity (in standard deviations) over background level crude Anscombe transform applied to data to stabilize the variance

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Fourier Bandpass Filter

Description

This is a plugin bundled with native ImageJ.

See IJ reference for more details > Link

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Chainer

Description

Chainer is a Python-based deep learning framework aiming at flexibility. It provides automatic differentiation APIs based on the define-by-run approach (a.k.a. dynamic computational graphs) as well as object-oriented high-level APIs to build and train neural networks. It also supports CUDA/cuDNN using CuPy for high performance training and inference. For more details of Chainer, see the documents and resources listed above and join the community in Forum, Slack, and Twitter.

QCAnet

Description

Quantitative Criterion Acquisition Network (QCA Net) performs instance segmentation of 3D fluorescence microscopic images. QCA Net consists of Nuclear Segmentation Network (NSN) that learned nuclear segmentation task and Nuclear Detection Network (NDN) that learned nuclear identification task. QCA Net performs instance segmentation of the time-series 3D fluorescence microscopic images at each time point, and the quantitative criteria for mouse development are extracted from the acquired time-series segmentation image. The detailed information on this program is described in our manuscript posted on bioRxiv.

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Manual tracking with TrackMate

Description

Manual tracking using Trackmate plugin (comes with FIji, so no installation required if you are using Fiji). 

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