Protein expression at the cellular level!

Understanding tissue heterogeneity is critical to answering key biological questions and a key concept in identifying clinically relevant association.

Our Different Techniques

GeoMx DSP uses fluorescently-labeled antibodies to define a region of interest (ROI). Antibodies labeled with unique photocleavable oligonucleotide are also applied on a 5 micron tissue section. Every selected region of interest is sequentially exposed to UV light to decouple the oligonucleotides from the profiling reagents. Decoupled oligonucleotides deposited into wells are  hybridized to Nanostring® barcodes and quantitated on the nCounter platform.

This non invasive approach allows the preservation of the tissue integrity, captures the tissue morphology and permits the high multiplexing of protein quantification –  up to 80 proteins in a run from a single tissue section.

Hyperion™ Imaging System (Imaging Mass Cytometry technique) performed on 5 micron tissue sections uses antibodies coupled to stable metal isotopes and then inserted into a chamber, where a pulsed UV laser disintegrates the tissue. The system allows the analysis of tissue sections with a resolution of 1 μm2, at a laser repeat rate of 100 Hz, creating individual plumes of particles for each point which will be transferred to the Helios mass cytometer. Individual metal isotopes are detected and indexed relative to the location of the source, providing information on the intensity and spatial distribution of antibodies in tissues using multiparametric visualization software.

The core advantage of Imaging Mass Cytometry compared to fluorescent microscopy is that there is almost no noise in the data as each metal isotopes has its on distinct detection peak and does not overlap with other metal isotopes. There is almost zero background noise and thus the contrast between markers of interest and the background is ideal for image analysis. Because of this lack of noise, a single tissue sample can be imaged for 40+ markers simultaneously from a single slide.

Applications

  • Spatially differentiated profiling of tumor driver and immune-oncology genes
  • Biomarker discovery related to location and signaling of specific cells
  • Pathways analysis across tissue structures and complexity
  • High dimensional phenotypic and functional analysis of single cells

Resources

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