Summary
A bioRxiv preprint describes REZCUE, a method for imaging archived tissue in three dimensions while preserving DNA, RNA and proteins for later molecular analysis. The authors report its use across commercial antibodies and in a multi-omic analysis of one archived breast cancer specimen.
A bioRxiv preprint describes REZCUE, a method designed to image archived tissue in three dimensions before molecular tests that consume or alter the sample. The authors say it enables multiplexed immunofluorescence imaging of formalin-fixed, paraffin-embedded (FFPE) tissue while preserving DNA, RNA and proteins for later assays.
FFPE samples are widely used in pathology and can remain archived for decades. But preparing them for imaging can damage molecules needed for later analysis, while conventional section-based methods provide views of thin slices rather than the full tissue volume. REZCUE is intended to help researchers connect molecular measurements with the structure and location of cells in an intact specimen.
How REZCUE is intended to work
The method uses what the authors describe as depth-invariant antigen retrieval chemistry based on in situ protein refolding. Antigen retrieval helps antibodies recognise their targets in fixed tissue. REZCUE applies this step to support three-dimensional imaging, with the preprint reporting imaging to a depth of 4 mm.
The authors report restoring antigenicity in tissue archived for up to 30 years and using 160 commercial antibodies across tissue types. In multiplexed immunofluorescence, fluorescently labelled antibodies reveal selected proteins; using multiple antibodies allows several targets to be imaged in the same tissue. The preprint says imaging can be performed before destructive molecular profiling, providing a three-dimensional spatial reference for aligning later measurements with tissue architecture.
A multi-omic analysis of archived breast cancer
To demonstrate the approach, the researchers analysed a breast cancer specimen archived for 25 years. They combined REZCUE-guided three-dimensional imaging and clonal segmentation with spatial transcriptomics, mass spectrometry, miRNA sequencing and whole-genome sequencing. The authors report identifying a luminalB/HER2-positive clone that conventional section-based spatial-omics had missed in this specimen.
The result illustrates the method’s proposed value: archived tissue could be examined first as an intact structure, then profiled with several molecular techniques while retaining a spatial map for interpretation. This could make existing specimens more useful for studying how distinct cell populations are arranged within tissue.
The work is presented as a bioRxiv preprint. Its multi-omic demonstration centres on one archived breast cancer specimen, so the reported finding is an example of the method’s use rather than evidence of its clinical performance across patients.