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

Actin Treadmilling01:18

Actin Treadmilling

9.7K
Actin filaments undergo polymerization and depolymerization from either end. The polymerization and depolymerization rates depend on the cytosolic concentration of free G-actins. The polymerization rate is generally higher at the plus or barbed end, while the depolymerization rate is higher at the minus or pointed end. At a steady state, critical concentration describes the concentration of free G-actin monomers at which the polymerization rate at the plus end is equal to that of the...
9.7K
Introduction to Actin01:26

Introduction to Actin

6.6K
Actin is a highly conserved cytoskeletal protein found abundantly in eukaryotic cells. It constitutes 10% weight of the total cellular protein in muscle cells, while in non-muscle cells, it is lower and makes up around 1–5 percent of the total cell protein. Actin found in the unicellular amoebae and complex multicellular animals is around 80% similar, demonstrating their conservation over a billion years of evolution.  Actin coding genes are conserved within species and across...
6.6K
Actin Polymerization01:42

Actin Polymerization

8.6K
Actin polymerization occurs through the head-to-tail association of binding sites on monomeric actin or G-actin to form filamentous or F-actin. The polymerization can be divided into three phases ̶  nucleation, elongation, and steady-state phase.
The nucleation phase involves forming a stable nucleus consisting of three actin monomers to form a new actin filament. Actin-binding proteins such as formins and Arp2/3 complex help filament growth post-nucleation. The Formins form straight...
8.6K
Actin Filament Depolymerization01:19

Actin Filament Depolymerization

3.9K
Actin filaments (F-actin) are composed of actin subunits. The dissociation of actin monomers can occur from either end of F-actin. The rate of dissociation is faster from the minus-end or the pointed end, where the actin subunits exist with a bound ADP, together known as ADP-actin. The depolymerization of F-actin is aided by proteins, including the actin-depolymerizing factor (ADF) and cofilin family of proteins, gelsolin, and glia maturation factor (GMF).
In F-actin, the ADF/cofilin proteins...
3.9K
Formation of Higher-order Actin Filaments01:11

Formation of Higher-order Actin Filaments

3.6K
The polymerization of G-actin monomers into filamentous F-actin is a multi-step process. Once the F-actins are formed, they can bundle together in different arrangements to form higher-order networks and regulate cellular functions. Common examples include the formation of lamellipodia and filopodia at the cell's leading edge by actin reorganization in a migrating cell. The microvilli on the brush border epithelial cells are also formed through the F-actin network.
The high-order actin...
3.6K
The Role of Actin and Myosin in Non-muscle Cells01:10

The Role of Actin and Myosin in Non-muscle Cells

4.8K
Actin and myosin or actomyosin filaments also play a significant role in cells other than those involved in muscle contraction (which occurs within the sarcomere of muscle cells). The mechanism of non-muscle cell contractile bundles was first observed in Dictyostelium and Acanthamoeba. In non-muscle cells, two bundles are commonly found: stress fibers and actomyosin adherence belts. These contractile bundles are smaller and less organized than the ones found in muscle cells. They  are held...
4.8K

You might also read

Related Articles

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

Sort by
Same author

Tie2 inhibition disrupts TMEM doorway function and reduces dissemination in pancreatic ductal adenocarcinoma.

Journal of experimental & clinical cancer research : CR·2026
Same author

Morphometric analysis of the thymic epithelial cell network using integrated and orthogonal digital pathology approaches.

Communications biology·2026
Same author

Aging disrupts sympathetic innervation of the thymus.

Cell reports·2026
Same author

Corrections to "Nonlinear Imaging Histopathology: A Pipeline to Correlate Gold-Standard Hematoxylin and Eosin Staining With Modern Nonlinear Microscopy".

IEEE journal of selected topics in quantum electronics : a publication of the IEEE Lasers and Electro-optics Society·2026
Same author

Metastatic dissemination of breast cancer stem cells requires MenaINV for lung extravasation but not survival.

bioRxiv : the preprint server for biology·2026
Same author

Racial disparity in pro-metastatic tumor microenvironment in treatment naïve breast cancer.

NPJ breast cancer·2026

Related Experiment Video

Updated: Jan 27, 2026

Labeling F-actin Barbed Ends with Rhodamine-actin in Permeabilized Neuronal Growth Cones
09:14

Labeling F-actin Barbed Ends with Rhodamine-actin in Permeabilized Neuronal Growth Cones

Published on: March 17, 2011

15.2K

Optimizing leading edge F-actin labeling using multiple actin probes, fixation methods and imaging modalities.

Vera DesMarais1,2,3, Robert J Eddy1, Ved P Sharma1,3

  • 1Department of Anatomy & Structural Biology, Albert Einstein College of Medicine, 1300 Morris Park Ave, Bronx, NY 10461, USA.

Biotechniques
|March 15, 2019
PubMed
Summary

Alexa Fluor-488 Phalloidin offers superior actin cytoskeleton labeling and signal stability for visualizing dynamic lamellipodia in breast cancer cells. Other probes showed limitations in performance and specificity.

Keywords:
F-tractinLifeactactincell motilitylamellipodiaphalloidin

More Related Videos

Quantification of Filamentous Actin F-actin Puncta in Rat Cortical Neurons
10:13

Quantification of Filamentous Actin F-actin Puncta in Rat Cortical Neurons

Published on: February 10, 2016

12.8K
Study of the Actin Cytoskeleton in Live Endothelial Cells Expressing GFP-Actin
08:37

Study of the Actin Cytoskeleton in Live Endothelial Cells Expressing GFP-Actin

Published on: November 18, 2011

27.7K

Related Experiment Videos

Last Updated: Jan 27, 2026

Labeling F-actin Barbed Ends with Rhodamine-actin in Permeabilized Neuronal Growth Cones
09:14

Labeling F-actin Barbed Ends with Rhodamine-actin in Permeabilized Neuronal Growth Cones

Published on: March 17, 2011

15.2K
Quantification of Filamentous Actin F-actin Puncta in Rat Cortical Neurons
10:13

Quantification of Filamentous Actin F-actin Puncta in Rat Cortical Neurons

Published on: February 10, 2016

12.8K
Study of the Actin Cytoskeleton in Live Endothelial Cells Expressing GFP-Actin
08:37

Study of the Actin Cytoskeleton in Live Endothelial Cells Expressing GFP-Actin

Published on: November 18, 2011

27.7K

Area of Science:

  • Cell Biology
  • Biochemistry
  • Molecular Biology

Background:

  • Dynamic lamellipodia are crucial for cell migration in breast adenocarcinoma.
  • Accurate visualization of the actin cytoskeleton is essential for studying cell motility.
  • Existing actin probes vary in performance and reliability.

Purpose of the Study:

  • To systematically evaluate and compare commercial actin-filament probes for visualizing F-actin-rich lamellipodia.
  • To optimize protocols for actin cytoskeleton imaging in highly motile cancer cells.
  • To identify the most reliable probes for studying dynamic cellular structures.

Main Methods:

  • Evaluation of four Phalloidin-fluorophores, two anti-actin antibodies, and three live-cell actin probes.
  • Testing across five different fixation conditions and three imaging platforms.
  • Assessment of labeling efficiency, signal stability, and probe specificity.

Main Results:

  • Alexa Fluor-488 Phalloidin demonstrated the best overall actin cytoskeleton labeling and signal maintenance.
  • Anti-actin antibodies exhibited significant limitations across various fixation and permeabilization conditions.
  • Live-cell probes showed actin filament bias; TagRFP-Lifeact was excluded from lamellipodia, unlike mEGFP-Lifeact and F-tractin-EGFP.

Conclusions:

  • Alexa Fluor-488 Phalloidin is recommended for robust F-actin visualization in dynamic lamellipodia.
  • Standard antibody-based methods and some live-cell probes present challenges for accurate actin imaging.
  • Optimized probe selection is critical for reliable studies of cell migration and cytoskeleton dynamics.