COMET™ performs sequential immunofluorescence (seqIF™), which is a method that relies on repeated cycles of staining, imaging, and antibody elution, where elution is a gentle step to remove antibodies with a dedicated buffer and an optimized protocol.

The identification of the best staining conditions can be easily determined using an automated workflow on COMET™. A gentle but efficient elution step is performed after each staining cycle without affecting tissue morphology. Epitope stability can be assessed during panel optimization, enabling users to develop 40-plex panels with ease in a few automated steps.

This interactive dataset presents raw data to show that:

• An optimal elution efficiency (>95%) is achieved on COMET™: more than 40 antibodies have been successfully tested on COMET™ across multiple tissue types and species.
• Tissue integrity is well preserved throughout the cycles: seqIF™ assays consist of gentle, fast, and highly-controlled conditions that do not affect tissue morphology, even on delicate and fragile samples.
• Excellent epitope stability is reached across the whole assay from cycle 1 to cycle 20: on COMET™, 93% of tested markers show excellent stability independently of the tissue type, species, and preparation.

In this example, a human Formalin-Fixed Paraffin-Embedded (FFPE) tonsil section was used. Tonsil is a widely used control tissue to test immune cell markers. The tissue was stained on COMET™ for CD19, a membrane protein commonly used as a marker for B cells; DAPI was used to label the nuclei.

Users can navigate the file to explore the robustness of COMET™ on the following:

1. Elution efficiency
2. Tissue morphology preservation
3. Epitope stability

More information on this topic, additional data, and a description of the COMET™ workflow for the assessment of elution efficiency and epitope stability can be found in the detailed Technical Note here.