RNA imaging with smFISH enables quantitative and qualitative gene expression experiments within single cells. With smFISH, researchers can measure cellular mRNA and perform spatial localization and quantification in locations such as the cytoplasm and nucleus.

5 天之前 · The number of probes per RNA molecule significantly affects detection sensitivity. A typical smFISH experiment uses 20–50 oligonucleotides per target transcript, ensuring even low-abundance RNAs generate a detectable signal. While increasing probe numbers enhances fluorescence intensity, excessive probe density can cause steric hindrance ...

Single molecule fluorescence in situ hybridization (smFISH) is a powerful technique to study gene expression in single cells due to its ability to detect and count individual RNA molecules.

2022年10月24日 · Here, we present high-throughput smFISH (HT-smFISH), a simple and cost-efficient method for imaging hundreds to thousands of single endogenous RNA molecules in 96-well plates. HT-smFISH uses...

2018年5月25日 · Single molecule fluorescence in situ hybridization (smFISH) is a powerful technique to study gene expression in single cells due to its ability to detect and count individual RNA molecules.

2015年4月24日 · Single-molecule fluorescence in situ hybridization (smFISH) has emerged as a powerful tool for studying the copy number and spatial organization of RNAs in single cells either in isolation or in their native tissue context (2, 3).

2023年6月15日 · With our smFISH whole-mount protocol and image analysis pipeline, we can now quantitatively analyse mRNAs and proteins at the cellular and subcellular levels in plants.

As the power of smFISH method lies in the accurate detection of single mRNA molecules, it is important to perform proper controls for each new smFISH probe library generated. Here we outline a number of control experiments that address …

2024年3月20日 · FISH is a powerful technique which allows the visualization of mRNA molecules at the subcellular level in fixed samples, via probing with fluorescent oligos using widefield fluorescence microscopy.

2018年11月5日 · Here, we describe a single-molecule fluorescent in situ hybridization (smFISH) method (as performed in Haimovich et al., 2017 ) that detects single RNA molecules in individual cells.