寡核苷酸
化学
核酸
荧光显微镜
DNA
原位
背景(考古学)
显微镜
荧光
核糖核酸
原位杂交
核酸热力学
生物物理学
共轭体系
组合化学
生物化学
计算生物学
基因表达
基因
基序列
有机化学
古生物学
聚合物
物理
光学
生物
量子力学
作者
Gang Wen,Marisa Vanheusden,Volker Leen,Taoufik Rohand,Katy Vandereyken,Thierry Voet,Johan Hofkens
摘要
Expansion microscopy (ExM) enables the nanoscale imaging of ribonucleic acids (RNAs) on a conventional fluorescence microscope, providing information on the intricate patterns of gene expression at (sub)cellular resolution and within spatial context. To extend the use of such strategies, we examined a series of multivalent reagents that allow the labeling and grafting of deoxyribonucleic acid (DNA) oligonucleotide probes in a unified approach. We show that the reagents are directly compatible with third-generation in situ hybridization chain reaction RNA FISH (fluorescence in situ hybridization) techniques while displaying complete retention of the targeted transcripts. Furthermore, we validate and demonstrate that our labeling method is compatible with multicolor staining. Through oligonucleotide-conjugated antibodies, we demonstrate excellent performance in ×4 ExM and ×10 ExM, achieving a resolution of ∼50 nm in ×10 ExM for both pre- and postexpansion labeling strategies. Our results indicate that our multivalent molecules enable the rapid functionalization of DNA oligonucleotides for ExM.
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