Building block	Specification/details
Images	The image module is used to compile the images for analysis.
Metadata	The Metadata module allows to extract information describing your images (i.e., metadata).
NamesAndTypes	The NamesAndTypes module allows to assign a meaningful name to each image by which other modules will refer to it.
IdentifyPrimaryObjects	The nuclei are identified based on Hoechst staining. Nuclei at the border of the image are removed as well as objects not matching the size and intensity criteria (threshold).
IdentifyPrimaryObjects	The TMRM staining is identified. Objects not matching the size and intensity criteria (threshold) are excluded.
IdentifySecondaryObjects	Cell edges are defined based on the previously selected nuclei, here defined as “Cell”.
RelateObjects	TMRM objects (child object) are associated with the previously defined Cell objects (parent object). This enables to calculate mean measurement values for TMRM associated to the Cell.
MeasureImageAreaOccupied	The area occupied by previously identified objects is measured (DNA, Cells and TMRM).
MeasureObjectIntensity	TMRM intensity is calculated in relation to the cell. Additional intensity parameters can be chosen (e.g., intensity of the nuclei).
MeasureObjectSizeShape	Additional output parameter for size and shape of the objects can be chosen.
ExportToSpreadsheet	Output results and storage location are defined.