From LipidomicsWiki

Contents 1 P01 - Regensburg 1.1 Analysis of fluorescence high-content imaging data with the granularity application module of MetaXpress 2.0 1.1.1 Output parameters 2 P10 - MPI Dresden

P01 - Regensburg

Analysis of fluorescence high-content imaging data with the granularity application module of MetaXpress 2.0

Backgrounds of images analyzed with the granularity application module of MetaXpress 2.0 are processed using the Adaptive Background CorrectionTM system. This system automatically corrects uneven image background throughout the image by adapting to local content. This allows for more robust segmentation and analysis repeatability and enables to specify the detection sensitivity as intensity above local background. A gray level value is selected that represents the least bright granule minus the background intensity near the granule. The software estimates the background for each lipid droplet locally to correct for cases of images with uneven background. The same procedure was applied for the nuclei and the value of the background is subtracted from the value of the nucleus. To differentiate nuclei from non-nuclear material the appropriate minimum and maximum lipid droplet width in microns (µm) is selected. Lipid droplets less than the specified minimum width will be considered as noise patterns. In addition also the appropriate minimum and maximum nuclei width were selected to select the nuclei for segmentation.

 

Lipid droplet and nuclei segmentation to get cell-based data

Mean values for well-based data are generated

 

Output parameters

This application module detects, counts and measures granularity and nuclei characteristics and provides well and cell based data.

The following parameters are detected:

Lipid droplets per cell (average) – represents the number of all lipid droplets in one well related to the number of nuclei in this well

Mean lipid droplet area - demonstrates the mean lipid droplet area of all lipid droplets in one well related to the number of nuclei in this well

Average lipid droplet intensity - adds together the average intensity of pixels within the stained area in all cells of one well (4 fields per well) and averages them

For graphing the mean values of these parameters from each well were further calculated into mean values of those wells with the same treatment.

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P10 - MPI Dresden

Thiele_Report_dataprocess_12months.pdf