HCR RNA-FISH: Protocol Overview
The same 2-stage protocol is used independent of the number of target RNAs.
All major
automation platforms
Read-Out/Read-In
The read-out/read-in analysis framework enables bidirectional quantitative protein and RNA discovery in an anatomical context.
Read-Out/Read-In: How It Works
Quantitative Protein and RNA Imaging
By quantifying protein and RNA targets while preserving anatomical context, quantitative HCR™ IF + HCR™ RNA-FISH enables bidirectional quantitative discovery. ​​
Read-Out from anatomical space to expression space to discover quantitative co-expression relationships in selected regions of the specimen.
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Read-In from expression space to anatomical space to discover those anatomical locations in which selected quantitative co-expression relationships occur.
Read-Out from a Whole-Mount Zebrafish Embryo
The expression scatter plot below illustrates read-out from a whole-mount zebrafish embryo using subcellular quantitative HCR™ RNA-FISH, providing quantitative snapshots of gene co-expression during somite formation and maturation.
Amplitude and Slope
Quantitative subcellular voxel intensities shaded by anatomical region reveal changes in expression ratio (slope) and abundance (amplitude) for mRNAs tpm3 and myod1 during somite formation (PSM) and maturation (S10→S9→S8→S7).