The field of diagnostics is an advanced field with many identified tools for the detection and quantification of specific proteins. You can visit www.bosterbio.com/services/assay-services/ihc-histology-services to get the best Antibody IHC Optimization Service.
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Diagnosis in pathology or clinical structures presents many challenges due to time constraints, increased roughness, and complexity of detection methods, smaller samples, etc.
Pathological detection is most often carried out using antibody-antigen interactions (by precipitation of the visible product or detection based on fluorochromes).
The standard approach is to perform one or two protein tests simultaneously on serial sections, which is impractical in terms of low sample availability and large proteins to detect.
More often, fluorescent reporters are used for detection and quantification methods, such as flow cytometers, imaging cytometers, etc. using live tissue or lightly fixed tissue, whereas conventional immunohistochemistry involves highly fixed tissue sections.
After the development of user-friendly and numerous analyzes of formalin-fixed, paraffin-embedded (FFPE) samples, recently samples have been developed and pooled, known as multiplexing or multiplexing immunohistochemistry (mIHC), which allows the use of more than three spots.
The primary strategy for multiplexing IHC in the same region at the same time requires the use of numerous animal-specific secondary antibodies associated with dissimilar reporters against antibodies raised in dissimilar animals.
Although there are a limited number of antibody sources, it is certain that it is difficult to develop a common multiplexing strategy for staining antibodies developed on the same species with different colors. However, three approaches are possible:
First bleach the conjugated primary antibody directly, then add a set of antibodies.
The labeled antibody epitope blocks, followed by the second round of staining
Remove pre-labeled antibodies from sections after staining and imaging, followed by another set of labels.
Topography multiplexing can take new forms with the outline of the "upcoming Generation IHC" isotope signature and in-situ mass spectrometry detection.