Hybridization chain reaction enables a unified approach to multiplexed, quantitative, high-resolution immunohistochemistry and <i>in situ</i> hybridization
Maayan Schwarzkopf, Mike C. Liu, Samuel J. Schulte, Rachel Ives, N. Husain, Harry M. T. Choi, Niles A. Pierce
Abstract
RNA in situ hybridization based on the mechanism of the hybridization chain reaction (HCR) enables multiplexed, quantitative, high-resolution RNA imaging in highly autofluorescent samples, including whole-mount vertebrate embryos, thick brain slices and formalin-fixed paraffin-embedded tissue sections. Here, we extend the benefits of one-step, multiplexed, quantitative, isothermal, enzyme-free HCR signal amplification to immunohistochemistry, enabling accurate and precise protein relative quantitation with subcellular resolution in an anatomical context. Moreover, we provide a unified framework for simultaneous quantitative protein and RNA imaging with one-step HCR signal amplification performed for all target proteins and RNAs simultaneously.