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

Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

2.0K
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...
2.0K

You might also read

Related Articles

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

Sort by
Same author

Clinical and pulmonary function-based identification of HRCT-confirmed rheumatoid arthritis-associated interstitial lung disease.

Clinical rheumatology·2026
Same author

Developmental and behavioral toxicity of citalopram in zebrafish early life stages: Neurochemical alterations, oxidative stress responses, and transcriptomic signatures.

Aquatic toxicology (Amsterdam, Netherlands)·2026
Same author

A Spin-5/2 Triangular-Lattice Antiferromagnet Exhibiting Field-Driven Competing Magnetic Phases.

Journal of the American Chemical Society·2026
Same author

Quantum criticality enhanced millikelvin magnetic refrigeration in a large-spin-7/2 triangular lattice antiferromagnet.

Innovation (Cambridge (Mass.))·2026
Same author

Adamts4 coordinates the transcriptomic profile of primary rat costal chondrocytes.

Stem cells translational medicine·2026
Same author

Simvastatin Restores Uteroplacental Hemodynamics and Trophoblast Function in Obstetric Antiphospholipid Syndrome in a Placenta-on-a-Chip Model.

Advanced healthcare materials·2026

Related Experiment Video

Updated: May 10, 2025

Author Spotlight: Evaluation of Protein-Condensate Dynamics in Live Human Cells
06:48

Author Spotlight: Evaluation of Protein-Condensate Dynamics in Live Human Cells

Published on: January 5, 2024

3.1K

Label-Free Quantification of Protein Density in Living Cells.

Robert J Clements1, Ruixin Guo2, Jonathan C Petruccelli3

  • 1Department of Biological Science, Kent State University, Kent, Ohio.

Current Protocols
|April 24, 2025
PubMed
Summary

This study introduces a method to measure intracellular protein concentration in living cells using transport-of-intensity equation (TIE) imaging and transmission-through-dye (TTD) for volume determination, enabling research into macromolecular crowding effects.

Keywords:
cell volumemacromolecular crowdingprotein concentrationrefractive indextransmission‐through‐dye microscopytransport‐of‐intensity equation

More Related Videos

Proteome-wide Quantification of Labeling Homogeneity at the Single Molecule Level
08:29

Proteome-wide Quantification of Labeling Homogeneity at the Single Molecule Level

Published on: April 19, 2019

6.1K
How to Quantify the Fraction of Photoactivated Fluorescent Proteins in Bulk and in Live Cells
11:03

How to Quantify the Fraction of Photoactivated Fluorescent Proteins in Bulk and in Live Cells

Published on: January 7, 2019

6.6K

Related Experiment Videos

Last Updated: May 10, 2025

Author Spotlight: Evaluation of Protein-Condensate Dynamics in Live Human Cells
06:48

Author Spotlight: Evaluation of Protein-Condensate Dynamics in Live Human Cells

Published on: January 5, 2024

3.1K
Proteome-wide Quantification of Labeling Homogeneity at the Single Molecule Level
08:29

Proteome-wide Quantification of Labeling Homogeneity at the Single Molecule Level

Published on: April 19, 2019

6.1K
How to Quantify the Fraction of Photoactivated Fluorescent Proteins in Bulk and in Live Cells
11:03

How to Quantify the Fraction of Photoactivated Fluorescent Proteins in Bulk and in Live Cells

Published on: January 7, 2019

6.6K

Area of Science:

  • Cell biology
  • Biophysics
  • Microscopy

Background:

  • Intracellular water content and protein concentration are critical for cell health.
  • Cellular stress can lead to dehydration and increased protein concentration, potentially causing apoptosis.
  • Macromolecular crowding (MC) effects, influenced by protein concentration, are crucial for cellular metabolism and signaling but remain poorly understood.

Purpose of the Study:

  • To develop and validate a method for measuring intracellular protein concentration in living cells.
  • To enable quantitative studies on the biological roles and regulation of macromolecular crowding.
  • To provide experimental guidelines and tools for implementing the measurement technique.

Main Methods:

  • Utilizing transport-of-intensity equation (TIE) imaging to derive refractive index from defocused brightfield images.
  • Employing transmission-through-dye (TTD) for accurate cell volume determination.
  • Developing an ImageJ plugin for TIE image processing and providing protocols for sample preparation, image acquisition, and calibration.

Main Results:

  • Demonstrated a method for simple and accurate measurement of protein concentration in adherent cells based on refractive index.
  • TIE and TTD methods are shown to be compatible with each other and with fluorescence imaging.
  • The approach is adaptable for measuring protein concentration in subcellular organelles with modified volume determination.

Conclusions:

  • The developed TIE and TTD imaging approach provides a robust method for quantifying intracellular protein concentration.
  • This technique facilitates the investigation of macromolecular crowding's impact on cellular functions.
  • The provided ImageJ plugin and protocols lower the barrier for researchers to study cellular protein dynamics.