Pathology Image Analysis (PIA)
Applications
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| Nuclear, cytoplasmic, membranous |
Tissue microarray | Microvessel density | Nuclear cell counting | Ploidy analysis |
Rare event detection | |
Ploidy analysis
- Feulgen stained tissue samples (7 microns thick) are scanned at 100x (Note that the amount of Feulgen stain is directly proportional to the amount of DNA present in the nucleus) (Huang Q et al, BMC Clin Pathol. 2005 and 2008)
- The ACIS uses an inbuilt system that sets the filter threshold controlling the inclusion of stained nuclei at predefined (1–5) levels. The operator selects the level that helps the user separate/cut in between adjacent cells and cannot be identified by the software as individual cells, ranging from the least (level 1) to the most (level 5) aggressive.
- The system “remembers” analysis regions to prevent the user from re-collecting nuclei from the same region.
- To avoid inclusion of excessively overlapped nuclei, each of the cell separation profiles was designed to recognize and exclude nuclei that exhibit excessive overlapping based on signature combinations of size, shape, color and morphometric filter descriptors (Watershed segmentation algorithm).
- Overlapping nuclei, nuclear debris and other artifacts that escaped auto-detection and removal by the system were deleted by the operator.
- Qualified nuclei of approximately 30 control stromal cells, such as endothelial cells, macrophages, and fibroblasts, in the same tissue and about 200-targeted epithelial cells are obtained.
- DNA content histograms are automatically plotted in another window using the ACIS DNA ploidy software.
- The user can easily navigate between individual nuclei and their exact position on the DI histogram. The mean integrated optical density (IOD) of control cells is assigned a DI of 1, which serves as an internal diploid (2N) standard and reference for DI calculation of the targeted cells. The IOD of control cells has a coefficient of variation (CV) of less than 10%. The histograms of the target cells showed a primary G0/G1 peak, and additional cells with or without well defined peaks with DI outside the G0/G1 peak (e.g., in S- or G2 phase of the cell cycle).
- Aneuploidy is diagnosed when the targeted epithelial nuclei showed no diploid G0/G1 peak and the DI of the G0/G1 peak was clearly positioned outside the diploid range.
