New Approaches in Quantitative Digital Pathology for Cancer IHC measurement Research and Companion Diagnostic Development

Flagship Biosciences has recently launched a tumor immunology review portal that leverages the growing field of whole-slide imaging (WSI). With the application of digital analysis algorithms to quantitate image analysis data, enhanced accuracy and reproducibility are achieved with WSI. Furthermore, quantitative digital pathology and WSI allow for the objective quantitation of inflammatory cells in the tumor microenvironment providing an accurate determination of an IHC measurement.

Rather than manually assessing cell populations on a limited, subjective scale, WSI can be performed and statistically powerful data generated. In our CLIA-certified histology laboratory, Flagship has identified and validated key pairs of antibodies. This allows for the ability to stain and quantify multiple cell populations in a single section as is typically necessary with limited clinical biopsies. This technique can be utilized to evaluate patient populations as well as to evaluate treatment-derived effects on inflammatory cells.

Flagship Biosciences has been using this technology to quantify a variety of inflammatory cells (e.g., CD3⁺, CD4⁺,CD8⁺, FoxP3⁺, CD68⁺, CD163⁺, etc.) as well as quantify cells with specific inflammatory cell products (e.g., IL-1, IL-17, TNF, IFNγ, etc.) using our proprietary CellMap^TM algorithm. For example, CD8⁺ (Figure 1) and CD3⁺ (Figure 2) cells can be identified in colorectal cancer which can further be quantified using CellMap^TM (Figure 3).

Immunohistochemistry provides evidence of the distribution of CD8⁺ and CD3⁺ in colorectal cancer biopsies, but quantitative digital pathology allows one to quantitate these cells relative to the pathology and can provide a path to accurately determining the IHC measurement. Our extensive quantitative image analysis team can assist you at all stages of your research ranging from early discovery to the development of a tissue-based companion diagnostic.

Figure 1 (A, B, C). Labeling of CD8⁺ T cells in colorectal cancer (CRC).

In Figure 1A, CD8⁺ T cells (brown) are clearly shown (high powered field) in both the lamina propria as well as in the epithelial cell layer. In Figure 1B, using CellMap^TM CD8⁺ cells are selectively identified and quantified in the lamina propria. In Figure 1C, using CellMap^TM the region of interest is switched to the epithelial layer and CD8⁺ cells have been identified and quantified

A
B
C

Figure 2 (A and B). Quantifying CD3⁺ T cells in colorectal cancer (CRC).

In both Figure 2A and 2B (low power fields containing highlighted region of interest boxes), the CellMap^TM algorithm was employed to identify and quantify the CD3⁺ T cells (red) in either the epithelium (Figure 2A) or the connective tissue compartments (Figure 2B, lamina propria and submucosa).

A
B