HCR™ Amplification Technology
HCR™ Amplifiers enable 1-step, multiplex, quantitative, isothermal, room-temperature, high-gain, enzyme-free signal amplification
HCR™ Amplifier: How It Works
HCR™ Amplifier
HCR™ Amplifiers leverage dynamic nanotechnology to enable small amplification components to first penetrate a biological sample without interacting and then autonomously grow bright amplification polymers at the site of targets within the sample, enabling deep sample penetration and unmatched performance, versatility, and robustness.
An HCR™ Amplifier comprises two kinetically trapped reporter-labeled hairpins, h1 and h2, that co-exist metastably on lab time scales. Hairpins h1 and h2 store the energy to drive a conditional self-assembly cascade upon exposure to initiator i1. Probes that bind the target use initiator i1 to trigger a chain reaction of alternating h1 and h2 polymerization steps leading to growth of a reporter-decorated HCR™ amplification polymer tethered to the target.
HCR™ introduced the concept of conditional nucleic acid self-assembly, demonstrating that nucleic acid molecules can be engineered so as not to interact when mixed, instead self-assembling only upon exposure to a cognate initiator sequence. Conditional nucleic acid self-assembly now provides a conceptual cornerstone for the emerging field of dynamic nucleic acid nanotechnology.
Straightforward Multiplexing
HCR™ Amplifiers enable straightforward multiplexing using 1-step quantitative signal amplification for all targets simultaneously.
Automatic Background Suppression
Because HCR™ hairpins are kinetically trapped, they will not generate amplified background even if they bind non-specifically in the sample, providing automatic background suppression to dramatically enhance performance and ease-of-use.
Deep Sample Penetration
HCR™ Amplifiers are small hairpins that penetrate thick samples to execute an enzyme-free amplification cascade at the site of the target.
Orthogonal Amplifiers
The gel above demonstrates multiplex signal amplification by 4 orthogonal amplifiers (488, 546, 647, 700). In the absence of initiators, there is negligible leakage out of the metastable hairpin state. Introduction of any single initiator into the amplifier mixture triggers the corresponding amplifier.
Quantitative Amplified Signal
HCR™ provides quantitative signal amplification in diverse products:
A new era for in situ hybridization:
multiplex, quantitative, high-resolution, sensitive, versatile, robust
Experience multiplexed, quantitative, high-resolution immunfluorescence: amplified, enzyme-free, FFPE, simple
Quantitative 1-step enzyme-free HCR™ Gold signal amplification for all RNA and protein targets simultaneously