HCR RNA-FISH: Protocol Overview
The same 2-stage protocol is used independent of the number of target RNAs.
All major
automation platforms
HCR™ IF + HCR™ RNA-FISH Technology
Simultaneous multiplexed, quantitative, high-resolution protein and RNA imaging
HCR™ IF probes, amplifiers, and buffers and HCR™ RNA-FISH probe sets, amplifiers, and buffers enable a unified approach to multiplexed, quantitative, high-resolution immunofluorescence (IF) and RNA fluorescence in situ hybridization (RNA-FISH).
HCR™ IF + HCR™ RNA-FISH: How It Works
1. Protein Detection Stage
2. RNA Detection Stage
3. Amplification Stage
1. Protein Detection Stage
Protein targets are detected with unlabeled primary antibody probes and initiator-labeled secondary antibody probes.
2. RNA Detection Stage
RNA targets are detected with split-initiator DNA probes that colocalize full initiator i1 only when they bind specifically to the target.
3. Amplification Stage
Initiators trigger self-assembly of tethered fluorescent amplification polymers for all protein and RNA targets simultaneously.
Quantitative 1-step enzyme-free signal amplification for all protein and RNA targets simultaneously
Simple Robust Protocols
HCR™ IF + HCR™ RNA-FISH protocols are simple, robust, and enzyme-free, requiring only 3 stages independent of the number of protein and RNA targets.
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Straightforward Multiplexing
HCR™ enables straightforward multiplexing using 1-step quantitative signal amplification for all protein and RNA targets simultaneously.
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Any organism across the tree of life
Free custom probe design for any target mRNA in any organism. For protein targets, plug-and-play using your own 1º antibodies of choice (without modification) in combination with validated HCR™ 2º antibody probes.
Simultaneous Quantitative High-Resolution Protein and RNA Imaging
Subcellular protein and RNA imaging enables protein and RNA relative quantitation with subcellular resolution in the anatomical context of highly autofluorescent samples.
The same 3-stage enzyme-free protocol is used independent of the number of protein and RNA targets
Protein and RNA relative quantitation with subcellular resolution in the anatomical context of highly autofluorescent samples
