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Related Experiment Video

Updated: Nov 3, 2025

High Resolution Fluorescent In Situ Hybridization in Drosophila Embryos and Tissues Using Tyramide Signal Amplification
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Highly Sensitive and Multiplexed In Situ RNA Profiling with Cleavable Fluorescent Tyramide.

Lu Xiao1, Joshua Labaer1, Jia Guo1

  • 1Biodesign Institute & School of Molecular Sciences, Arizona State University, Tempe, AZ 85287, USA.

Cells
|June 2, 2021
PubMed
Summary
This summary is machine-generated.

Researchers developed a sensitive and specific multiplexed RNA imaging method for analyzing RNA in various tissues, including challenging formalin-fixed paraffin-embedded (FFPE) samples. This new technique enhances in situ RNA profiling capabilities for complex biological systems.

Keywords:
FISHfluorescence in situ hybridizationgenomicstranscriptomicstranscripts

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Area of Science:

  • Molecular Biology
  • Genomics
  • Biotechnology

Background:

  • Understanding biological systems necessitates tools for quantifying multiple transcripts in their native cellular locations.
  • Current multiplexed RNA imaging technologies face limitations in sensitivity and specificity, particularly in autofluorescent tissues like FFPE.

Purpose of the Study:

  • To develop a highly sensitive and specific multiplexed in situ RNA profiling approach.
  • To overcome limitations of existing methods for analyzing RNA in challenging tissue types.

Main Methods:

  • Utilizes target-specific oligonucleotide probes in pairs for initial hybridization.
  • Employs signal amplification via oligonucleotide hybridization and horseradish peroxidase (HRP) with cleavable fluorescent tyramide (CFT).
  • Incorporates cyclic RNA staining, imaging, signal cleavage, and probe stripping for multiplexed analysis.

Main Results:

  • Demonstrates high sensitivity and specificity in multiplexed in situ RNA profiling.
  • Successfully applied the approach to fixed, frozen, and formalin-fixed paraffin-embedded (FFPE) tissues.
  • Enables profiling of diverse RNA species at optical resolution.

Conclusions:

  • The developed method offers a significant advancement for in situ RNA analysis.
  • It is effective across various tissue preservation states, including FFPE.
  • This technique expands the possibilities for studying complex biological systems at the transcriptomic level.