The Morphometry Fluo algorithm enables cells to be quantified using luminosity thresholds and morphomathematical operators. This algorithm is frequently used for IF quantifications and analysis of the colocalization of fluorescent staining.
The protocol :
The different stages of image analysis using the Morphometry Fluo algorithm are as follows :
- Image segmentation
Luminance segmentation is done on each of the fluorochromes.
- Application of morphomathematical operators
Analysis is refined by applying morphomathematical criteria such as smoothing, erosion and dilation, watershed, size, etc.
- Analysis of co-staining
This analysis is carried via the intersection of masks.
Results present the number of segmented objects, the surface area occupied by the objects and include a description of each object.
It is also possible to construct a learning model based on the RFT tool. The IOBL imaging analysis algorithm is then used for the fluorescence-based quantification of multiplexed slides.