Linux

Super-resolution optical fluctuation imaging (SOFI) achieves 3D super-resolution by computing temporal cumulants or spatio-temporal cross-cumulants of stochastically blinking fluorophores. In contrast to localization microscopy, SOFI is compatible with weakly emitting fluorophores and a wider range of blinking conditions. Balanced SOFI analyses several cumulant orders for extracting molecular parameter maps, such as the bright and dark state lifetimes, the concentration and the brightness distributions of fluorophores within biological samples. In combination with a deconvolution of the cumulant images, the estimated parameter maps proved useful to balance the image contrast and to linearize the brightness and blinking response. Thereby, the image quality and the resolution were improved significantly.

The MorphoLeaf application allows you to extract the contour of multiple leaf images and identify their biologically-relevant landmarks. These landmarks are then used to quantify morphological parameters of individual leaves and to reconstruct average leaf shapes. MorphoLeaf is developed by the Modeling and Digital Imaging and the Transcription Factors and Architecture teams of the Institut Jean-Pierre Bourgin, INRA Versailles, France, and the Biophyscis and Development group at RDP, Lyon.

Free-D is a three-dimensional (3D) reconstruction and modeling software. It allows to generate, process and analyze 3D point and surface models from stacks of 2D images. Free-D is an integrated software, offering in a single graphical user interface all the functionalities required for 3D modeling. It runs on Linux, Windows, and MacOS. Free-D is developed by the Modeling and Digital Imaging team of the Institut Jean-Pierre Bourgin, INRA Versailles, France.

SuReSim (Super Resolution Simulation) is an open-source simulation software for Single Molecule Localization Microscopy (SMLM). The workflow of the SuReSim algorithm starts from a ground truth structure and lets the user choose to either directly simulate 3D localizations or to create simulated *.tiff-stacks that the user can analyze with any given SMLM reconstruction software. A 3D structure of any geometry, either taken from electron microscopy, designed de-novo from assumptions or known structural facts, is fluorophore-labeled in silico. A defined set of parameters is used to calculate and visualize the 3D localizations of the corresponding labels. The software package is accompanied with a library of model structures that can be imported and simulated. Users manual with tutorial provided.

Free-D (http://free-d.versailles.inra.fr/) is a 3D reconstruction and modeling software. It is multiplatform, free (but not open source) tool for academic research and teaching.

Here is how to proceed, using Free-D:

1. Segmentation:

* load (a collection of) individual 3d stacks

* (optional for serial sections) perform a 2D registration to align image slices

* segment/reconstruct 3D contours using snakes

* segment 3D spots

2. Construct average cell:

* normalize the contours to compute a average cell, by registering/warping 3D contours/surfaces

3. Quantification:

* project each individual cell to the average one

* build density maps to analyze (cartography)

A few notes for current software version (till 10/2016):

* input file format: tiff (not able to import bioformats)

* currently results are saved in customized format, but there is an exportor to convert this format into fiji readable one

* import already generated contours is on the software's TODO list