Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Ribosome Profiling02:24

Ribosome Profiling

3.7K
Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...
3.7K
FISH - Fluorescent In-situ Hybridization02:07

FISH - Fluorescent In-situ Hybridization

21.9K
Fluorescence in situ hybridization, or FISH, was developed in the early 1980s and has quickly become one of the most widely used techniques in cytogenetics. Labeled probes are used to bind complementary DNA or RNA sequences on a chromosome or in a region within a cell. Earlier, the probes could only be obtained by cloning or reverse transcription of a DNA template. Currently, the probe oligonucleotides can be synthesized synthetically. Additionally, with the advancement of optical techniques,...
21.9K
Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

2.3K
Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
2.3K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Identification of superstalsis, a motility program distinct from peristalsis in the Drosophila midgut.

Cell reports·2026
Same author

Editorial: Technologies for RNA Detection.

Bio-protocol·2026
Same author

SAMJ: fast image annotation on ImageJ/Fiji via segment anything model.

Nature communications·2026
Same author

Protocol to quantify glioblastoma cell invasion and nuclear deformations in 3D hydrogels.

STAR protocols·2026
Same author

Temporal dynamics of neuroplasticity and neurodegeneration in the central auditory system following noise-induced hearing loss: a multimodal imaging and histological study.

Acta neuropathologica communications·2026
Same author

Paired transcriptomics of glioblastoma peripheral and core areas reveals CNTN2 as a potential therapeutic target.

Cancer letters·2026

Related Experiment Video

Updated: Sep 25, 2025

Robust 3D DNA FISH Using Directly Labeled Probes
12:16

Robust 3D DNA FISH Using Directly Labeled Probes

Published on: August 15, 2013

34.9K

Interrogating RNA and protein spatial subcellular distribution in smFISH data with DypFISH.

Anca F Savulescu1, Robyn Brackin2, Emmanuel Bouilhol3,4

  • 1Division of Chemical, Systems & Synthetic Biology, Institute for Infectious Disease & Molecular Medicine, Faculty of Health Sciences, University of Cape Town, 7295 Cape Town, South Africa.

Cell Reports Methods
|April 27, 2022
PubMed
Summary

DypFISH quantitatively analyzes subcellular RNA and protein localization using single-molecule FISH and immunolabeling. This method reveals molecular spatial patterns and their biological significance in various cell types.

Keywords:
RNA subcellular localizationRipley's Kimage analysismicrofabricated patternssingle-molecule FISH

More Related Videos

Author Spotlight: Investigating mRNA Spatial Distribution in Drosophila Muscle Tissue
10:22

Author Spotlight: Investigating mRNA Spatial Distribution in Drosophila Muscle Tissue

Published on: September 8, 2023

1.7K
Use of Single Molecule Fluorescent In Situ Hybridization SM-FISH to Quantify and Localize mRNAs in Murine Oocytes
08:18

Use of Single Molecule Fluorescent In Situ Hybridization SM-FISH to Quantify and Localize mRNAs in Murine Oocytes

Published on: April 24, 2019

10.9K

Related Experiment Videos

Last Updated: Sep 25, 2025

Robust 3D DNA FISH Using Directly Labeled Probes
12:16

Robust 3D DNA FISH Using Directly Labeled Probes

Published on: August 15, 2013

34.9K
Author Spotlight: Investigating mRNA Spatial Distribution in Drosophila Muscle Tissue
10:22

Author Spotlight: Investigating mRNA Spatial Distribution in Drosophila Muscle Tissue

Published on: September 8, 2023

1.7K
Use of Single Molecule Fluorescent In Situ Hybridization SM-FISH to Quantify and Localize mRNAs in Murine Oocytes
08:18

Use of Single Molecule Fluorescent In Situ Hybridization SM-FISH to Quantify and Localize mRNAs in Murine Oocytes

Published on: April 24, 2019

10.9K

Area of Science:

  • Cellular and Molecular Biology
  • Biophysics
  • Genomics

Background:

  • Single-cell RNA sequencing identifies cellular subtypes by transcript counts.
  • Cellular differences extend to the spatial distribution of molecules, including RNA.
  • Understanding subcellular localization is crucial for cell function.

Purpose of the Study:

  • To present DypFISH, a novel approach for quantitative analysis of subcellular RNA and protein localization.
  • To introduce analytical techniques for interrogating single-molecule RNA fluorescence in situ hybridization (smFISH) and protein immunolabeling data.
  • To explore molecular clustering, mRNA-protein localization relative to cellular structures, and spatial distribution interdependencies.

Main Methods:

  • Development of DypFISH for quantitative subcellular localization analysis.
  • Integration of single-molecule RNA fluorescence in situ hybridization (smFISH) with protein immunolabeling.
  • Application of analytical tools to study molecular clustering and spatial relationships.
  • Utilizing cell micropatterning to control cellular architecture and reduce localization variation.

Main Results:

  • DypFISH enables quantitative investigation of RNA and protein subcellular localization.
  • Analytical tools characterize molecular clustering and mRNA-protein spatial distribution patterns.
  • Cell micropatterning facilitated the reduction of localization variation, aiding pattern characterization.
  • The method was successfully applied to physiological systems, including skeletal muscle fibers.

Conclusions:

  • DypFISH provides a powerful quantitative framework for studying subcellular molecular organization.
  • The analytical approach reveals insights into the spatial dynamics of RNA and protein within cells.
  • This method is applicable to diverse biological systems, advancing our understanding of cellular function.