核糖核酸
多路复用
计算生物学
原位
杠杆(统计)
滚动圆复制
计算机科学
转录组
生物
原位杂交
信使核糖核酸
基因表达
DNA
化学
基因
遗传学
人工智能
聚合酶
电信
有机化学
作者
Ian Dardani,Benjamin Emert,Yogesh Goyal,Connie Jiang,Amanpreet Kaur,Jasmine Lee,Sara H. Rouhanifard,Gretchen M. Alicea,Mitchell Fane,Min Xiao,Meenhard Herlyn,Ashani T. Weeraratna,Arjun Raj
出处
期刊:Nature Methods
[Springer Nature]
日期:2022-10-24
卷期号:19 (11): 1403-1410
被引量:14
标识
DOI:10.1038/s41592-022-01653-6
摘要
RNA labeling in situ has enormous potential to visualize transcripts and quantify their levels in single cells, but it remains challenging to produce high levels of signal while also enabling multiplexed detection of multiple RNA species simultaneously. Here, we describe clampFISH 2.0, a method that uses an inverted padlock design to efficiently detect many RNA species and exponentially amplify their signals at once, while also reducing the time and cost compared with the prior clampFISH method. We leverage the increased throughput afforded by multiplexed signal amplification and sequential detection to detect 10 different RNA species in more than 1 million cells. We also show that clampFISH 2.0 works in tissue sections. We expect that the advantages offered by clampFISH 2.0 will enable many applications in spatial transcriptomics. ClampFISH 2.0 enables highly specific multiplexed signal amplification in RNA FISH. The approach was used to detect 10 RNA species that ranged in abundance in more than 1 million cells and is also applicable to tissue sections.
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