time-series

The workflow measures the growth of cells in 3D, combining an ImageJ macro for preprocessing and successive tracking using Imaris.  

The sample dataset (available in the github repository) contains 2-Photon images of neurons. The neurons were imaged in 3D at two time frames.To allow measuring significant differences in cell volume, the time gap between the frames is large (ca. 30 min) and the animal was removed in the waiting phase. For this reason, there is a considerable shift in sample position between the frames that has to be corrected before cell detection and tracking.

The workflow consists of following steps:

1. Import of single tiff slices [imageJ macro]

2. Organizing the data in a 4D time series with 2 time frames [imageJ macro]

3. Correction of shift between the time frames by rigid registration [imagJ macro]

4. Bleaching correction [imageJ macro]

5. Export of preprocessed image data in ics/ids format [imageJ macro]

6. Import of ics/ids data to Imaris [Imaris]

7. Cell object detection as "Imaris Surface Object" [Imaris]

8. Tracking cell objects over time [Imaris]

9. Split Tracks (use Imaris XT extension "Split Tracks") to generate single cell objects [Imaris]

10. Export the statistics: Select the complete folder, go to the statistics tab and use ‚Full Export’ [Imaris]

The preprocessing macro can be referenced here.

The sample images were acquired by Cordula Ulbrich (Petzold Group at German Center of Neurodegenerative Disesases (DZNE), Bonn, Germany).

Input data type: tiff

Output data type: data table

u-Track is a client-side OMERO MATLAB application implementing the sophisticated multiple-particle tracking algorithm of Jaqaman et al. . It works on data previously imported into an OMERO server, and produces results in the form of MATLAB data structures as well as providing functionality to visualise these results.

This simple KNIME workflow solution tracks 2D objects/cells in time series. After a few intensity based preprocessing steps, objects/cells are segmented first, then it uses Fiji TrackMate LAP method for the tracking task.

Documentation starts from p23 of the linked PDF. 

Example Image: mitocheck_small.tif (2.9M)

The article describes how a FRAP experiment can be conducted and subsequently analyzed. This includes steps in ImageJ and subsequent normalization of the intensity data.

This is a qualitative analysis, and curve fitting is done using Excel. 

Requires "Template matching and Slice alignment plugin"

OMERO.figure is an OMERO web application that makes generating figures from images in an OMERO image database very quick and easy. The images in the figure link back to the original data, greatly simplifying the process of adjusting the view and keeping track of original data. PDF documents are generated, which can be opened using e.g. Adobe Illustrator or Inkscape in order to produce the final finished figure.