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MedicineMedicine and Medical Research

Multiplex Quantitative Histologic Analysis of Human Breast Cancer Cell Signaling and Cell Fate

Authors: William M. Lee; Badrinath Roysam; PENNSYLVANIA UNIV PHILADELPHIA
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.

Limitations: APPROVED FOR PUBLIC RELEASE
Description: Final rept. 1 May 2007-30 Apr 2010
Pages: 51
Report Date: MAY 2010
Contract Number: W81XWH-07-1-0325
Report Number: A269435
Keywords relating to this report:
ANTIGENS
BIOLOGICAL STAINS
BREAST CANCER
CELLS(BIOLOGY)
COMPUTER PROGRAMS
CYTOLOGY
DISCOLORATION
DISTRIBUTION
HISTOLOGICAL TECHNIQUES
HISTOLOGY
HORMONES
HUMANS
IMMUNOLOGY
MARKERS
MICROSCOPY
MOLECULES
MULTIPLEXING
MULTISPECTRAL
NEOPLASMS
NUCLEI
QUANTITATIVE ANALYSIS
RECEPTOR SITES(PHYSIOLOGY)
SEGMENTED
SEPARATION
SIGNALS
SPECTRA
STRUCTURAL PROPERTIES
TRACER STUDIES
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