Cleavable Fluorescent Probe (CFP™)

An innovative technology empowers spatial biology

Spatomics employs a multiplexing strategy centered around the Cleavable Fluorescent Probe (CFP™), which allows iterative cycles of staining, imaging, and fluorophore cleaving.

In each cycle, multiple protein targets are first recognized by primary antibodies from different species of varied immunoglobulin classes. Subsequently, the first target is stained by secondary antibodies and the cleavable fluorescent tyramide (CFT). HRP catalyzes the coupling reaction between CFT and the tyrosine residues on the endogenous proteins in close proximity. After imaging, the fluorophores are gently cleaved with nearly 100% efficiency and HRP is simultaneously deactivated. Through reiterative analysis cycles, many distinct proteins can be quantitatively profiled in single cells in-situ.

CFP cycling workflow

Antibody compatibility

The CFP™ assay allows users to use their preferred standard antibodies without labelling.

Significantly enhanced sensitivity

Leverage the efficient tyramide signal amplification (TSA) system, CFP™ offers consistent performance with the conventional TSA assays. CFP™ is compatible with FFPE tissues.

Efficient fluorophore cleavage without loss of protein antigeneity

Unlike conventional TSA assays, with CFP™ fluorescence signals can be efficiently removed using a gentle process, preserving the protein antigenicity for reliable cycling.

Intact tissue integrity throughout the cycles

CFP™ assays are mild and fast cycling process, ensuring minimal impact on tissue morphology.

Automation friendly

The CFP™ assay is automation friendly. There are both a manual version and a fully automated version, accessible through Spatomics’ proprietary system

Due to its efficient fluorophore cleavage and preservation of protein antigenicity throughout the staining cycles, CFP enables users to easily create high-multiplex panels with their preferred antibodies. This example shows a tonsil tissue stained through 28 cycles.

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