Multiplex Quantitative Histologic Analysis of Human Breast Cancer Cell Signaling and Cell Fate
Abstract
Many molecular events and cellular processes are preserved in fixed human tumor specimens, and access to this information awaits a method for them to be quantified and analyzed. Hormone receptors, HER2, cell signaling and proliferation events are prognostically and therapeutically important in human breast cancer and can be revealed by immunohistological staining. A novel platform for study of immunostained breast cancer specimens is being developed that will quantify analyte antigens on a cellular basis, i.e. cytometrically. The platform uses multispectral microscopy to examine breast cancer specimens that have been immunostained for multiple cell type and analyte antigens using different chromogens and fluorophores. Multispectral microscopy and spectral separation permits staining for individual antigens to be distinguished and separated from staining for other antigens in multiplex-stained slides. Stains for structural and cell type antigens (e.g. nuclei, epithelial cytokeratins, E-cadherin) are used by FARSIGHT software to segment individual nuclei and cells in images and to identify those that are breast cancer cells. Stains for biomarkers and cell signaling antigens (e.g. ER, PR, HER2. Ki67, p-ERK, p-AKT) are then associated with the segmented cells to quantify expression of these analytes in breast cancer cells on a per-cell basis. Progress in the past year has developed the cell segmentation function of FARSIGHT software so that nuclear (ER, PR, Ki67, p-ERK, p-AKT) and cytosolic/cell membrane analytes (HER2, p-ERK, p-AKT, p-S6) can be cytometrically quantified and their subcellular distribution determined in human breast cancer specimens.
Document Details
- Document Type
- Technical Report
- Publication Date
- May 01, 2010
- Accession Number
- ADA534962
Entities
People
- Badrinath Roysam
- William M. Lee
Organizations
- University of Pennsylvania