Applications Across Biology

Subcellular Resolution, Multiple Assays

Spatial Context Brings Biological Insight

A holistic understanding of cells in their native tissues is key to unlocking new discoveries in neuroscience, cancer, infectious disease, immunology and developmental biology.

The Rebus Esper is a single-instrument solution designed to deliver high-throughput spatial data at subcellular resolution from a multitude of assays.

The first assay from Rebus Biosystems is the Esper High Fidelity assay for RNA analysis in fresh frozen mouse and human tissues.

Additional assays will be released soon, all taking advantage of the resolution, scale and speed of the Rebus Esper.

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Esper High Fidelity Assay

Robust, reliable gene expression analysis:

  • Single molecule resolution
  • High sensitivity and specificity
  • No barcoding or enzymatic amplification
  • Access cytoplasm and nucleus
  • Up to 30 custom genes

Multiplexed, quantitative data for:

  • Cell mapping
  • Validating scRNA-seq results
  • Detecting rare cells
  • Exploring genes with low expression
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Every Transcript, Every Cell

The Esper High Fidelity assay enables multiplex analysis of gene expression with no compromise in sensitivity or specificity.  Detect every transcript in every cell with confidence.

We’ve taken an assay long-recognized as the gold-standard for gene expression analysis, single-molecule in situ fluorescence hybridization (smFISH), and made it plug and play for all researchers.

 

Custom Assays

With the Esper High Fidelity assay, you can analyze up to 30 custom genes. Kits from Rebus Biosystems contain all necessary probes and reagents.

An optimized on-instrument tissue pretreatment protocol allows high-quality data to be acquired from samples, regardless of endogenous background fluorescence or lipofuscin content.

 

High sensitivity and Specificity

A high level of multiplexing is achieved without the use of barcoding or enzymatic amplification. A probe tiling strategy ensures signal is seen only from true targets.

The result is the ability to resolve and map individual RNA molecules in their native locations in tissues across a wide range of expression levels with the highest possible detection efficiency and specificity.

 

Target Accessibility

Whether the target is an mRNA molecule in the cytoplasm or a long non-coding RNA or nascent RNA within cell nucleus, the Esper High Fidelity assay shows consistent detection capability.

Applications

Cell Type Mapping

To arrive at the single cell tissue map shown in Figure 1, a set of 24 genes – 7 reference and just 17 cell type specific — were probed in a coronal section of mouse brain.

After clustering and manual annotation based on marker gene expression, cell types were mapped back to X,Y space.

The identified clusters clearly displayed the expected spatial organization and faithfully recapitulated the tissue architecture. Furthermore, the relative abundances of the different cell types precisely match previously published data.

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High-quality spatial data for cell type identification and mapping

Figure 1
figure1

Detecting Rare Cells

In Figure 2, a low expressing gene (CRYM) found in less than 1% of cells in a section of adult human brain is visualized.

The high throughput of the Rebus Esper enabled imaging of a large enough area to find these rare cells.

The high specificity of the Esper High Fidelity assay prevented false positives and ambiguous results, while the assay’s high sensitivity allowed detection of such low expressing targets.

Figure 2
figure2

scRNA-Seq Validation

In Figure 3, cell mapping data for a section of human fetal brain is shown.

This tissue was evaluated using the Esper High Fidelity assay for 30 genes first identified through scRNA-seq, which suggested a diversity of cell types across regions of the developing human cortex.

These data together enabled direct observation of area specific molecular diversity and allowed inference of functional roles of regionally diverse clusters based on known differentiation trajectories and migratory gradients.

Figure 3
Figure3_cellmap