Fluorescent In Situ Detection of Small RNAs in Plants Using sRNA-FISH

Kun Huang, Blake C. Meyers, Jeffrey L. Caplan

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

Abstract

Plant small RNAs are 21–24 nucleotide, noncoding RNAs that function as regulators in plant growth and development. Colorimetric detection of plant small RNAs was made possible with the introduction of locked nucleic acid probes. However, fluorescent detection of plant small RNAs has been challenging due to the high autofluorescence from plant tissue. Here we report a fluorescent in situ detection method for plant small RNAs. This method can be applied to most plant samples and tissue types and also can be adapted for single-molecule detection of small RNAs with super-resolution microscopy.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages101-111
Number of pages11
DOIs
StatePublished - 2024

Publication series

NameMethods in Molecular Biology
Volume2784
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Autofluorescence
  • Confocal
  • FISH
  • In situ hybridization
  • Linear spectral unmixing
  • Plant
  • Quantification
  • RNAs
  • SIM
  • STORM
  • Small RNAs
  • Spectra

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