Thursday, 3 August 2023

XFP to RNAscope Imaging Fiduciary Marker

DRAQ7™ has been demonstrated as a fiduciary/registration marker to allow co-alignment of sequential images using nuclei staining from (native) fluorescence and then RNAscope™ procedures on the same re-processed slice.

This workflow allowed for the successful protein detection prior to the RNA detection since the sample processing in the latter includes a protein degradation step that would be otherwise incompatible with detection of (in this case, fluorescent) protein.  DRAQ7 was applied to the fixed, permeabilized slice to reveal nuclei alongside the virally-transduced mBFP signals.  After recovery and re-processing for in situ hybridization, DRAQ7 was re-applied to the slice for nuclei detection with the RNAscope signals, again by fluorescence microscopy.

This was part of a work published in Nature from the lab of Chris Harvey (Professor of Neurobiology, Harvard Medical School) in the pursuit of the coordination and synchronisation of a specialised set of neurons involved in error-correction, specifically in navigation in the mouse model.

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Reference:

Green, J., Bruno, C.A., Traunmüller, L. et al. A cell-type-specific error-correction signal in the posterior parietal cortex. Nature (2023). https://doi.org/10.1038/s41586-023-06357-1

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