Figure 2.

(A) Workflow for performing quantitative digital pathology on 2D tissue sections using image analysis and stereology. Blue shading designates preanalytical factors, which are procedures a tissue undergoes before quantitative analysis. Gray shading designates generation of whole slide digital images, and green shading designates quantitative analysis of digital images. (B) Workflow for image analysis, a 2D analysis technique used to obtain quantitative data from tissue sections stained with histochemical stains, IHC, or ISH. (C) Workflow for stereology, a 3D analysis technique that is considered the gold standard for quantitative analysis of tissue sections. A critical preanalytical step often required when performing stereology is the need to obtain a reference volume, which is the volume of tissue, organ, or biopsy before processing of the tissue. Abbreviations: 2D, two-dimensional; 3D, three-dimensional; IHC, immunohistochemistry; ISH, in situ hybridization.