CSBDeep, a toolbox for Content-aware Image Restoration (CARE) in Knime

Description

Deep learning based restoration, with guidelines for training. See also the Fiji plugin.

CSBDeep, a toolbox for Content-aware Image Restoration (CARE) in Fiji

Description

Deep learning for fluorescence image restoration (denoising, deconvolution). Requires training on your data set but the procedure is described.

CARE

Real-time multi-view deconvolution

Description

In light-sheet microscopy, overall image content and resolution are improved by acquiring and fusing multiple views of the sample from different directions. State-of-the-art multi-view (MV) deconvolution simultaneously fuses and deconvolves the images in 3D, but processing takes a multiple of the acquisition time and constitutes the bottleneck in the imaging pipeline. Here, we show that MV deconvolution in 3D can finally be achieved in real-time by processing cross-sectional planes individually on the massively parallel architecture of a graphics processing unit (GPU). Our approximation is valid in the typical case where the rotation axis lies in the imaging plane.

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Multiview Reconstruction

Description

The Multiview Reconstruction software package enables users to register, fuse, deconvolve and view multiview microscopy images. The software is designed for lightsheet fluorescence microscopy (LSFM), but is applicable to any form of three or higher dimensional imaging modalities like confocal timeseries or multicolor stacks. 

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BigStitcher

Description

The BigStitcher is a software package that allows simple and efficient alignment of multi-tile and multi-angle image datasets, for example acquired by lightsheet, widefield or confocal microscopes. The software supports images of almost arbitrary size ranging from very small images up to volumes in the range of many terabytes, which are for example produced when acquiring cleared tissue samples with lightsheet microscopy.

ORION

Description

ORION: Online Reconstruction and functional Imaging Of Neurons: segmentation and tracing of neurons for reconstruction.

A project to develop tools that explore single neuron function via sophisticated image analysis. ORION software bridges advanced optical imaging and compartmental modeling of neuronal function by rapidly, accurately, and robustly generating, from structural image data, a cylindrical morphology model suitable for simulating neuronal function. The goal of this project is to develop a computational and experimental framework to allow real-time mapping of functional imaging data (e.g., spatio-temporal patterns of dendritic voltages or intracellularions) to neuronal structure, during the very limited duration of an acute experiment.

ORION_example_result

Super-Resolution Radial Fluctuations (SRRF)

Description

SRRF is a high-performance analytical approach for Live-cell Super-Resolution Microscopy, provided as a fast GPU-enabled ImageJ plugin. SRRF is capable of extracting high-fidelity super-resolution information from TIRF, widefield and confocals using conventional fluorophores such as GFP. SRRF is capable of live-cell imaging over timescales ranging from minutes to hours.

Authors: Ricardo Henriques' lab

Comparison TIRF - SRRF

ImmunoRatio

Description

Analyzing ER, PR, and Ki-67 immunohistochemistry

ImmunoRatio is an ImageJ plugin to quantify haematoxylin and DAB-stained tissue sections by measuring the percentage of positively stained nuclear area (labeling index), described in [bib]2452[/bib].

Notes for use:

  • It is important to read the URL instructions and original paper to understand what is being measured. In particular, the primary measurement made is percentage of the total nuclear area, not the percentage of detected nuclei (the latter being the more common method of assessing e.g. Ki67). This may be further modified by the Result correction equation.
  • Ultimately ImmunoRatio relies on thresholding (color deconvolved [bib]2451[/bib]) images to define 'nucleus' vs 'non-nucleus' regions according to staining intensity. Therefore dark artefacts, such as tissue folds, are likely to cause errors.
  • The pixel size is not read automatically from the image, but rather the source image scale should be entered into the dialog box - and the image rescaled accordingly prior to analysis. This scale value is the inverse of the value normally found for pixel width and pixel height under Image -> Properties... (i.e. pixel width & height are given in microns per pixel; the dialog box asks for pixels per micron).

Web application: ImmunoRatio

Example Image: Sample ImmunoRatio results

References

  1. [2452] Tuominen VJRuotoistenmäki SViitanen AJumppanen MIsola J.  2010.  ImmunoRatio: a publicly available web application for quantitative image analysis of estrogen receptor (ER), progesterone receptor (PR), and Ki-67.. Breast Cancer Res. 12(4):R56.
  2. [2451] Ruifrok ACJohnston DA.  2001.  Quantification of histochemical staining by color deconvolution.. Anal Quant Cytol Histol. 23(4):291-9.
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Quantifying staining in tissue sections

Description

The Color Deconvolution plugin for ImageJ can be used to digitally separate up to three stains from brightfield images, after which standard ImageJ commands can be used. The algorithm is described in Ruifork and Johnston (2001).

However, it is very important to take into consideration the caveats on the linked URL. In particular, note that:

  • Stain colors depend on numerous factors, such as the precise stains and scanner; therefore, the 'default' stain vectors (used to define the colors) are unlikely to be optimal and may be very inaccurate. See the URL instructions for how to create new stain vectors.
  • Pixel values should be interpreted with extreme caution; in particular, note the warning regarding 'brown' staining that attempting to quantify DAB intensity using this plugin is not a good idea.

Note, the pixel values provided by this plugin are 8-bit and not equivalent to 'optical densities' frequently presented in the literature.

Color deconvolution is particularly helpful in separating stains so that stained regions can be detected (e.g. by setting a threshold), and then the number or areas of stained structures may be quantified. Two potential approaches would be:

  1. If one measurement should be made for the entire image:
    • Image > Adjust > Threshold...
    • Edit > Selection > Create Selection
    • Analyze > Measure
  2. If distinct structures should be measured:
    • Image > Adjust > Threshold...
    • Analyze > Analyze Particles...
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AutoQuant

Description

Deconvolution software

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Image restoration with AutoQuant