Related Experiment Video
Updated: Jun 23, 2026
11:15
A Guide to Structured Illumination TIRF Microscopy at High Speed with Multiple Colors
Published on: May 30, 2016
Super-resolution video microscopy of live cells by structured illumination
Peter Kner1, Bryant B Chhun, Eric R Griffis
1Department of Biochemistry and Biophysics, San Francisco, California, USA.
Nature Methods
|May 1, 2009
Summary
High-speed structured-illumination microscopy achieves 100-nm resolution for live cell imaging. This breakthrough enables observing dynamic cellular processes at unprecedented speed and detail.
Area of Science:
- Cell biology
- Microscopy
- Biophysics
Background:
- Widefield fluorescence microscopy offers limited resolution.
- Structured-illumination microscopy (SIM) enhances resolution but is typically too slow for live imaging.
Purpose of the Study:
- To develop a high-speed structured-illumination microscope for dynamic live imaging.
- To achieve 100-nm resolution at high frame rates.
Main Methods:
- Development of a novel high-speed structured-illumination microscope.
- Utilizing total internal reflection mode for imaging.
- Video imaging of cellular dynamics in Drosophila melanogaster S2 cells.
Main Results:
- Achieved 100-nm resolution at frame rates up to 11 Hz.
- Successfully imaged cellular dynamics for several hundred time points.
- Demonstrated the capability for live imaging of tubulin and kinesin dynamics.
Conclusions:
- The developed high-speed SIM is suitable for dynamic live cell imaging.
- Enables visualization of subcellular processes with high spatial and temporal resolution.
- Opens new avenues for studying molecular motors and cytoskeletal dynamics in vivo.
Related Concept Videos
01:37Super-resolution Fluorescence Microscopy
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 developed.
01:28Three-Dimensional Microscopy in Microbiology
Three-dimensional imaging techniques are essential in cell biology, allowing researchers to visualize intricate cellular structures with high resolution. Two prominent methods, Differential Interference Contrast Microscopy (DIC) and Confocal Scanning Laser Microscopy (CSLM), provide distinct advantages for imaging live and thick specimens, respectively.Differential Interference Contrast MicroscopyDIC microscopy enhances contrast in transparent, unstained samples by converting phase...
01:16Confocal Fluorescence Microscopy
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,...