Description:
Executive Summary
A new method for immunohistochemical (IHC) staining allowing colocalization studies of six or more proteins within a single tissue section.
Background
Visual colocalization of molecular species within sectioned tissue can provide powerful insights into cellular biochemistry and often provides the basis for further study of protein-protein interactions. However, current methods are imperfect and are limited to at most three simultaneous probes. Multi-color fluorescence is limited by few non-overlapping combinations of fluorescent tags, antibody cross-reactivity, photobleaching and autofluorescence of paraffin-embedded tissue. Neither immunoenzyme multi-staining nor multiplex immunostaining chips can be used to view colocalization in the same cellular compartment.
Invention Description
An immunohistochemical method for multiple antigen colocalization accomplished by rounds of staining, imaging and antibody-stripping. This new method has successfully been employed for staining of up to six unique proteins on a single tissue section without loss of tissue antigenicity.
Advantages
- Colocalization of up to six antigens with IHC staining methods &bull
- Uses traditional IHC staining methods to which pathologists are accustomed and do not suffer from signal-to-noise limiting autoflourence
- Method verified on rat, mouse and human tissue samples
- Protocol can be implemented as a commercialized kit or within automated slide staining and scanning devices
Potential Applications
SIMPLE has a wide range of applications, including basic research in cell and molecular biology, translational research and diagnostics. The technique can be carried out manually, using automated slide stainers or high-throughput immunochemical assays. Image acquisition is greatly facilitated by digital microscope slide scanning systems.
