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.
Straightforward Multiplexing
HCR™ enables straightforward multiplexing using 1-step quantitative signal amplification for all protein and RNA targets simultaneously.
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
