Related Experiment Video
Updated: May 3, 2026
12:58
Fluorescence Recovery after Photobleaching of Yellow Fluorescent Protein Tagged p62 in Aggresome-like Induced Structures
Published on: March 26, 2019
16.2K
Nonlinear light-sheet fluorescence microscopy by photobleaching imprinting.
Liang Gao1, Liren Zhu, Chiye Li
1Optical Imaging Laboratory, Department of Biomedical Engineering, Washington University, , St Louis, MO 63130, USA.
Journal of the Royal Society, Interface
|January 31, 2014
Summary
We developed a new nonlinear light-sheet fluorescence microscopy (LSFM) technique using photobleaching imprinting. This method improves imaging contrast and optical sectioning for clearer visualization of biological samples.
Area of Science:
- Biophotonics and advanced microscopy techniques.
Background:
- Light-sheet fluorescence microscopy (LSFM) offers advantages for biological imaging.
- Existing LSFM methods face challenges with limited field of view and optical sectioning capabilities.
- Scattered light can degrade image quality and contrast in LSFM.
Purpose of the Study:
- To introduce a novel nonlinear light-sheet fluorescence microscopy (LSFM) scheme.
- To enhance imaging performance by improving field of view and optical sectioning.
- To reduce background noise and increase image contrast for better visualization.
Main Methods:
- Implementation of a nonlinear LSFM scheme utilizing photobleaching imprinting.
- Measurement of photobleaching-induced fluorescence decay for image analysis.
- Development of a method to simultaneously achieve a large imaging field of view and a thin optical section.
Main Results:
- Achieved simultaneous large field of view and thin optical sectioning.
- Significantly reduced scattered-light-induced background.
- Considerably improved image contrast.
Conclusions:
- The developed photobleaching imprinting LSFM scheme offers superior imaging performance.
- This technique is expected to broaden LSFM applications into the optical quasi-ballistic regime.
- Enables advanced studies on non-transparent biological samples.
Related Concept Videos
01:37Super-resolution Fluorescence Microscopy
12.3K
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...
12.3K
01:16Confocal Fluorescence Microscopy
16.0K
Confocal microscopy is an advanced microscopic technique. The prime advantage of the confocal microscope over other microscopy techniques is its ability to block the out-of-focus light from the illuminated samples using pinholes. It is widely used with fluorescence optics to obtain high-resolution, sharp contrast images. Unlike optical microscopes, confocal microscopes use a focused beam of light laser to scan the entire sample surface at different z-planes. These microscopes are, therefore,...
16.0K
01:19Protein Dynamics in Living Cells
1.9K
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...
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...
1.9K