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MULTIPLEX USE CASES

Immuno-Oncology

Get the full picture for your tumor microenvironment research

Immuno-oncology has been one of the primary drivers of the current multiplex technology development. Multiplex immunohistochemical (IHC) analysis of formalin-fixed, paraffin-embedded (FFPE) tissue samples allows researchers to study the spatial relationships between different cell phenotypes in situ. Tonsil is often the first step to check that the antibodies are identifying the correct immune cells. After that the actual tumor microenvironment can be employed with hematologic samples, staining often determines cell lineage/origin as well. This phased process of optimization is long but ensures fidelity of results.

(This is the battle field) Human tonsil, 4-plex assay
DAPI (Counterstain)
Spectrum Gold
CY7
CY5
Spectrum Green
2-plex FISH assay: green (CEP17) and orange (HER2 Gene)

Fluorescence In Situ Hybridization (FISH) assay

Nucleic Acids in Cancer research

FISH is a sensitive, accurate, and reliable technique widely applied in cancer research. The genetic defects uncovered by FISH represent early genetic triggers or events responsible for cancer at stem-cell level. FISH provides cell-based context for specific genomic aberrations and plays an important role in detecting specific biomarkers in solid and hematologic neoplasms

Quantitative Multicolor QM-FISH 

Pairs of probes have been conventionally used to detect a single genetic event like deletion or amplification of a locus or chromosomal translocation. However, with the discovery of multigenic diseases including cancer, simultaneous detection of such genes by using multiple probes on a single slide aids understanding of disease progression (quantitative multicolor FISH).

UroVysion Bladder cancer FISH kit (4 different chromosomes have been assayed: gold (9p21), aqua(CEP17), red(CEP3) & green(CEP7)
UroVysion Bladder cancer FISH kit analyzed with Aperio FISH A/D algorithm provides color visualization of the scoring classes of the FISH assay

Tissue Heterogeneity

It is easy to think of heterogeneity between tumor and non-tumor, but it is also well known that there is heterogeneity within the tumor itself. Not every cell will stain the same and not every marker will be present in the same cells. Multiplex IHC and ISH uncover this multi-faceted approach to tissue heterogeneity and provides context.

Example of breast cancer image using Aperio IA SW

Characterizing biomarkers within the tumor or the stroma" shall be placed below the images on the right side.