Research Article

Automated and Reproducible Detection of Vascular Endothelial Growth Factor (VEGF) in Renal Tissue Sections

Table 1

Pipeline programmed for image analysis with CP.

ModuleOperation

(1) LoadImagesIdentify and load images in .tiff
(2) ColorToGrayConversion method: split
(3) MorphOperation: invert
(4) IdentifyPrimaryObjects(a) Identify an object of interest: core
(b) Maximum and minimum area: 13-40
(c) Threshold strategy: adaptive
(d) Threshold method: MCT
(5) IdentifySecondaryObjects(a) Object name: cell
(b) Method to identify the secondary objects: propagation
(c) Threshold strategy: adaptive
(d) Threshold method: kapur
(6) IdentifyTertiaryObjectsObject name: cytoplasm
(7) MeasureObjectSizeShapeMeasurement object: cytoplasm
(8) MeasureObjectIntensityMeasurement object: cytoplasm

(1) Load user-defined images. (2) Convert the original images to grayscale images. (3) Invert intensities to have bright nuclei. (4) Identify the primary object of interest (in this case, the nucleus). (5) Identify the secondary object (in this case, the entire cell). (6) Create the tertiary object (cytoplasm) by subtracting the primary object from the secondary object, i.e., subtracting the nucleus from the cell. (7) Analyze morphologic parameters in the object called cytoplasm. (8) Calculate intensity parameters in the object called cytoplasm.