Quantum dot-based in situ hybridization and immunohistochemistry to detect mRNA and protein at subcellular levels, comparison with studies using electron microscopy

Abstract

Semiconductor nanocrystals (Quantum dots, Qdots) have advantages of narrow, symmetric emission spectra with multiple resolvable colors that can be excited simultaneously using a single excitation wavelength. Therefore, different sized Qdots can serve the simultaneous detection of mRNA and protein. The procedure of in situ hybridization (ISH) and immunohistochemistry (IHC) using confocal laser scanning microscopy (CLSM) and Qdots is simpler than combined ISH and IHC using electron microscopy. This method is applicable to the three-dimensional detection of several molecules including mRNA and proteins.