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

Phosphoinositides and PIPs01:42

Phosphoinositides and PIPs

Phosphoinositides are a group of phospholipids containing a glycerol backbone with two fatty acid chains and a phosphate attached to a myoinositol sugar ring. The inositol head group extends into the cytoplasm, where it is modified by adding phosphate groups to form phosphatidylinositol phosphates or PIPs.
Different phosphoinositides are synthesized and recruited on the cytosolic face of the plasma membrane. The localization of specific phosphoinositides concentrated in separate membrane...
Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

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...

You might also read

Related Articles

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

Sort by
Same author

RAB4A acts as a negative feedback regulator of extracellular vesicle secretion during TGF-β signaling.

The FEBS journal·2026
Same author

The Welander TIA1 mutation dedifferentiates insulin-producing cells: Reversal by a GLP-1 receptor agonist.

The Journal of biological chemistry·2026
Same author

Loss of ErbB3 redirects Integrin β1 from early endosomal recycling to secretion in extracellular vesicles.

The Journal of cell biology·2025
Same author

Somatostatin triggers local cAMP and Ca<sup>2+</sup> signaling in primary cilia to modulate pancreatic β-cell function.

The EMBO journal·2025
Same author

Mechanistic insights and approaches for beta cell regeneration.

Nature chemical biology·2025
Same author

Real-time detection of somatostatin release from single islets reveals hypersecretion in type 2 diabetes.

Acta physiologica (Oxford, England)·2025

Related Experiment Video

Updated: Jun 10, 2026

Single-molecule Super-resolution Imaging of Phosphatidylinositol 4,5-bisphosphate in the Plasma Membrane with Novel Fluorescent Probes
07:26

Single-molecule Super-resolution Imaging of Phosphatidylinositol 4,5-bisphosphate in the Plasma Membrane with Novel Fluorescent Probes

Published on: October 15, 2016

Imaging phosphoinositide dynamics in living cells.

Anne Wuttke1, Olof Idevall-Hagren, Anders Tengholm

  • 1Department of Medical Cell Biology, Uppsala University, Uppsala, Sweden.

Methods in Molecular Biology (Clifton, N.J.)
|July 21, 2010
PubMed
Summary

This study presents methods for visualizing phosphoinositide dynamics in single cells using fluorescent protein probes. These techniques enable high-resolution imaging of inositol lipid changes crucial for cellular signaling.

More Related Videos

Radiolabeling and Quantification of Cellular Levels of Phosphoinositides by High Performance Liquid Chromatography-coupled Flow Scintillation
10:52

Radiolabeling and Quantification of Cellular Levels of Phosphoinositides by High Performance Liquid Chromatography-coupled Flow Scintillation

Published on: January 6, 2016

PIP-on-a-chip: A Label-free Study of Protein-phosphoinositide Interactions
10:58

PIP-on-a-chip: A Label-free Study of Protein-phosphoinositide Interactions

Published on: July 27, 2017

Related Experiment Videos

Last Updated: Jun 10, 2026

Single-molecule Super-resolution Imaging of Phosphatidylinositol 4,5-bisphosphate in the Plasma Membrane with Novel Fluorescent Probes
07:26

Single-molecule Super-resolution Imaging of Phosphatidylinositol 4,5-bisphosphate in the Plasma Membrane with Novel Fluorescent Probes

Published on: October 15, 2016

Radiolabeling and Quantification of Cellular Levels of Phosphoinositides by High Performance Liquid Chromatography-coupled Flow Scintillation
10:52

Radiolabeling and Quantification of Cellular Levels of Phosphoinositides by High Performance Liquid Chromatography-coupled Flow Scintillation

Published on: January 6, 2016

PIP-on-a-chip: A Label-free Study of Protein-phosphoinositide Interactions
10:58

PIP-on-a-chip: A Label-free Study of Protein-phosphoinositide Interactions

Published on: July 27, 2017

Area of Science:

  • Cell Biology
  • Biochemistry
  • Microscopy

Background:

  • Inositol lipid derivatives are vital for cellular signaling and processes.
  • Measuring phosphoinositide changes in real-time within single cells is essential for understanding their roles.
  • Specific protein domains interacting with inositol lipids can be engineered as cellular probes.

Purpose of the Study:

  • To develop and present protocols for fluorescence imaging of phosphoinositide dynamics in living cells.
  • To enable high spatial and temporal resolution of phosphoinositide concentration changes.
  • To visualize various phosphoinositide species in different subcellular compartments.

Main Methods:

  • Utilizing protein domains tagged with green fluorescent protein variants as phosphoinositide probes.
  • Employing total internal reflection fluorescence microscopy for time-lapse imaging of the plasma membrane.
  • Applying these methods to study phospholipase C- and PI3-kinase-induced inositol lipid changes in insulin-secreting cells.

Main Results:

  • Demonstrated successful visualization of phosphoinositide dynamics in single living cells.
  • Showcased the utility of total internal reflection fluorescence microscopy for capturing rapid inositol lipid fluctuations.
  • Quantified changes in inositol lipids induced by specific signaling enzymes.

Conclusions:

  • The presented fluorescence imaging protocols offer high spatial and temporal resolution for studying phosphoinositide metabolism.
  • These techniques are adaptable for investigating spatio-temporal regulation of phosphoinositides across various cell types.
  • This work enhances the understanding of inositol lipid roles in cellular signaling pathways.