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

2.7K
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...
2.7K
Super-resolution Fluorescence Microscopy01:37

Super-resolution Fluorescence Microscopy

14.6K
Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been...
14.6K

You might also read

Related Articles

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

Sort by
Same author

Call For Papers: Molecular Understanding and Formulation Design for Peptide Delivery.

Molecular pharmaceutics·2026
Same author

Analyzing Spatial Variations in Molecular Mobility in Hydrated Amorphous Drug-Polymer Blends Using Fourier Transform Fluorescence Recovery After Photobleaching and Image Segmentation.

The journal of physical chemistry. B·2026
Same author

The expanding role of formulations to enable oral delivery of poorly water-soluble drugs.

Nature reviews. Drug discovery·2026
Same author

Addressing the Release and Permeation Challenges of High-<i>T</i><sub><i>g</i></sub> Drugs in Amorphous Solid Dispersions.

Molecular pharmaceutics·2026
Same author

Investigating the Relationship between In Vitro and In Vivo Performance: The Role of Drug Loading, Release Rate, and Surface Area.

Molecular pharmaceutics·2026
Same author

Role of Dissolution Medium pH in the Release Behavior of Basic Drug-Copovidone Amorphous Solid Dispersions.

Molecular pharmaceutics·2026

Related Experiment Video

Updated: Feb 17, 2026

A Guide to Structured Illumination TIRF Microscopy at High Speed with Multiple Colors
11:15

A Guide to Structured Illumination TIRF Microscopy at High Speed with Multiple Colors

Published on: May 30, 2016

26.2K

Pixel-Wise Diffusion Imaging by Structured Illumination Microscopy Fluorescence Recovery after Photobleaching

Dustin M Harmon1, Maura M Wimsatt1, Michelle L Cousineau2

  • 1Department of Chemistry, Purdue University, West Lafayette, Indiana 47907, United States.

Analytical Chemistry
|February 16, 2026
PubMed
Summary
This summary is machine-generated.

Structured illumination microscopy- fluorescence recovery after photobleaching (SIM-FRAP) enables pixel-wise diffusion mapping in amorphous solid dispersions. This technique quantifies molecular mobility across dissolution fronts, revealing insights into formulation design.

More Related Videos

Internalization and Observation of Fluorescent Biomolecules in Living Microorganisms via Electroporation
15:27

Internalization and Observation of Fluorescent Biomolecules in Living Microorganisms via Electroporation

Published on: February 8, 2015

17.5K
Fluorescence Recovery after Photobleaching of Yellow Fluorescent Protein Tagged p62 in Aggresome-like Induced Structures
12:58

Fluorescence Recovery after Photobleaching of Yellow Fluorescent Protein Tagged p62 in Aggresome-like Induced Structures

Published on: March 26, 2019

16.0K

Related Experiment Videos

Last Updated: Feb 17, 2026

A Guide to Structured Illumination TIRF Microscopy at High Speed with Multiple Colors
11:15

A Guide to Structured Illumination TIRF Microscopy at High Speed with Multiple Colors

Published on: May 30, 2016

26.2K
Internalization and Observation of Fluorescent Biomolecules in Living Microorganisms via Electroporation
15:27

Internalization and Observation of Fluorescent Biomolecules in Living Microorganisms via Electroporation

Published on: February 8, 2015

17.5K
Fluorescence Recovery after Photobleaching of Yellow Fluorescent Protein Tagged p62 in Aggresome-like Induced Structures
12:58

Fluorescence Recovery after Photobleaching of Yellow Fluorescent Protein Tagged p62 in Aggresome-like Induced Structures

Published on: March 26, 2019

16.0K

Area of Science:

  • Pharmaceutical Sciences
  • Materials Science
  • Microscopy

Background:

  • Quantifying molecular diffusivity in amorphous solid dispersions (ASDs) is crucial for designing effective drug formulations.
  • High drug loadings in ASDs can reduce dissolution rates due to phase separation, forming drug-rich barrier layers.
  • Molecular mobility is key to understanding dissolution trends across different phases within a compact.

Purpose of the Study:

  • To develop and apply a novel SIM-FRAP technique for pixel-wise diffusion mapping in pharmaceutical compacts.
  • To investigate molecular mobility across the dissolution front of ASDs, including glassy, hydrated, gel, and phase-separated domains.
  • To demonstrate the capability of SIM-FRAP for segmentation-free, quantitative analysis of molecular diffusion.

Main Methods:

  • Integration of structured illumination microscopy (SIM) reconstruction algorithms with spatial Fourier transform fluorescence recovery after photobleaching (FT-FRAP).
  • Utilizing SIM for photobleaching followed by spatial Fourier transformation and inverse Fourier transformation to generate modulation-dependent amplitude maps.
  • Performing pixel-wise fits to exponential decay functions to determine diffusion coefficients across the entire field of view.

Main Results:

  • SIM-FRAP successfully generated pixel-wise diffusion maps of molecular mobility across the dissolution front of model pharmaceutical compacts.
  • The technique revealed smoothly varying diffusivity in compact and gel phases, alongside discontinuous changes in phase-separated domains.
  • A large number of diffusion coefficients (up to ~1 million) were recovered per experiment, providing detailed spatial information.

Conclusions:

  • SIM-FRAP offers a powerful, segmentation-free method for quantifying molecular diffusivity in ASDs.
  • The technique directly visualizes spatial variations in molecular mobility, consistent with hypothesized phase separation phenomena.
  • The experimental simplicity of SIM-FRAP facilitates its broader application in formulation development and materials characterization.