HCR v3.0
A new era for in situ hybridization
✓ Simple
✓ Amplified
✓ Multiplexed
✓ Quantitative
✓ Penetrating
✓ Resolved
✓ Sensitive
✓ Portable
✓ Versatile
✓ Compatible
✓ Robust
2-stage enzyme-free protocol
Signal elevated above autofluorescence
Simultaneous 1-step amplification for up to 5 target RNAs
Signal scales linearly with target abundance
Whole-mount vertebrate embryos and thick tissue sections
Subcellular or single-molecule resolution as desired
Single molecules detected in thick autofluorescent samples
Suitable for use with diverse targets in diverse organisms
Multiple assay formats: RNA-FISH, RNA flow cytometry, northern blots
Combine with tissue clearing as desired
Automatic background suppression throughout the protocol
Multiplexed, quantitative, sensitive, versatile, robust

Thick Autofluorescent Samples
Third-generation in situ hybridization chain reaction (HCR) redefines the state-of-the-art for RNA fluorescence in situ hybridization (RNA-FISH) in thick autofluorescent samples including whole-mount vertebrate embryos.
Straightforward Multiplexing
Orthogonal HCR amplifiers enable 1-step signal amplification for up to 5 target RNAs simultaneously. Learn more
Automatic Background Suppression
HCR v3.0 reagents provide automatic background suppression throughout the protocol, ensuring that even if probes or amplifiers bind non-specifically in the sample they will not generate amplified background, dramatically enhancing performance and ease-of-use. Learn more
Deep Sample Penetration
HCR amplification hairpins are short predominantly double-stranded oligos that penetrate thick samples prior to executing an enzyme-free amplification cascade at the site of the target.
Two Quantitative Imaging Modes
qHCR Imaging
Analog RNA relative quantitation with subcellular resolution in an anatomical context. Learn more
dHCR Imaging
Digital RNA absolute quantitation with single-molecule resolution in an anatomical context. Learn more
Simple Robust Protocols
HCR v3.0 protocols are simple, robust, and enzyme-free, requiring only 2 stages independent of the number of target RNAs. Learn more
Any Organism Across the Tree of Life
No matter what your sample (whole-mount embryo, thick brain slice, FFPE tissue section, model or non-model organism, mammalian cells, bacterial cells, multi-species environmental sample), we are happy to help! View Gallery