Histones, well known as packing components of DNA, have been extensively studied because their modifications are closely related to epigenetic processes. Thus, it is very important to map the position and distribution of the histone modifications for elucidating the underlying mechanism behind. However, conventional histone imaging methods are performed by using organic fluorescent dyes combined with secondary antibodies, has limitations in multiplexed and high-spatial imaging. To overcome these, optically tunable nanoparticles, such as gold nanoparticles (GNPs) and quantum dots (QDs) have recently drawn attentions as excellent imaging probes owing to their high-spatial resolution and multiplexing modalities. In this study, we develop novel imaging probes using GNPs and QDs conjugated with primary antibodies of histone proteins to visualize the distribution of the modified histone in a single nucleus. As conceptual demonstration, different trimethylation level and distribution of histone 3 lysine 9 (H3K9) in normal and senescence induced cells is clearly visualized by using GNPs and QDs. Furthermore, we demonstrate multi-functional imaging probes consist of both GNPs and Raman active fluorescent probes, which make it possible to apply to multiple imaging apparatus including dark-field microscopy, fluorescence microscopy, and Raman microscopy for histone mapping. The proposed imaging probes provide a powerful way to study not only histone codes, also other cellular targets such as organelles and biomolecules.

High-spatial single cell histone mapping using optically tunable nanoparticles

2019 Fall National Meeting

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