Phase imaging and synthetic aperture super-resolution via total internal reflection microscopy
Optics Letters
|May 2, 2018
Summary
Total internal reflection microscopy can be adapted into a phase microscope by detecting interference signals. This label-free technique offers improved resolution through synthetic aperture imaging, enhancing cell membrane visualization.
Area of Science:
- Biophysics
- Optical Microscopy
- Cell Biology
Background:
- Total internal reflection microscopy (TIRFM) is the standard for imaging fluorescent proteins near cell membranes.
- Current TIRFM primarily operates in fluorescence mode, limiting label-free imaging capabilities.
Purpose of the Study:
- To demonstrate TIRFM's potential as a label-free phase microscope.
- To enhance resolution in label-free microscopy techniques.
Main Methods:
- Detection of the coherent signal from interference between scattered and evanescent fields in TIRFM.
- Utilizing multiple illumination angles for synthetic aperture reconstruction.
Main Results:
- Total internal reflection microscopy can function as a phase microscope without fluorescence labeling.
- Synthetic aperture reconstructions yield higher resolutions than conventional label-free methods.
Conclusions:
- TIRFM offers a versatile, label-free phase contrast imaging modality.
- The developed technique significantly improves resolution for imaging cellular structures.
More Related Videos
10:19Visualizing Adhesion Formation in Cells by Means of Advanced Spinning Disk-Total Internal Reflection Fluorescence Microscopy
Published on: January 21, 2019
7.0K
14:09High-Throughput Total Internal Reflection Fluorescence and Direct Stochastic Optical Reconstruction Microscopy Using a Photonic Chip
Published on: November 16, 2019
7.4K
Related Concept Videos
01:05Total Internal Reflection Fluorescence Microscopy
13.5K
Total internal reflection fluorescence microscopy or TIRF is an advanced microscopic technique used to visualize fluorophores in samples close to a solid surface with a higher refractive index, such as a glass coverslip. TIRF only allows fluorophores in proximity to the solid surface to be excited. When light from a medium with a lower refractive index (such as air) hits the glass coverslip at a critical angle, the light undergoes total internal reflection stead of passing through the glass.
13.5K
01:37Super-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
01:13Attenuated Total Reflectance (ATR) Infrared Spectroscopy: Overview
1.3K
Attenuated total reflectance (ATR) infrared spectroscopy is a powerful analytical technique used to study the composition of materials. It is widely employed in chemistry, materials science, forensic science, and other fields where sample characterization is required. ATR has several advantages over traditional transmission IR spectroscopy, including the requirement of little to no sample preparation and the ability to analyze a wide range of samples.
The ATR process begins by directing a beam...
The ATR process begins by directing a beam...
1.3K
02:31Phase Transitions
23.3K
Whether solid, liquid, or gas, a substance's state depends on the order and arrangement of its particles (atoms, molecules, or ions). Particles in the solid pack closely together, generally in a pattern. The particles vibrate about their fixed positions but do not move or squeeze past their neighbors. In liquids, although the particles are closely spaced, they are randomly arranged. The position of the particles are not fixed—that is, they are free to move past their neighbors to...
23.3K
02:00Internal Energy
36.8K
The total of all possible kinds of energy present in a substance is called the internal energy (U), sometimes symbolized as E. Suppose a system with initial internal energy, Uinitial, undergoes a change in energy (transfer of work or heat), and the final internal energy of the system is Ufinal. Change in internal energy equals the difference between Ufinal and Uinitial.
36.8K
01:31Internal Receptors
74.7K
Many cellular signals are hydrophilic and therefore cannot pass through the plasma membrane. However, small or hydrophobic signaling molecules can cross the hydrophobic core of the plasma membrane and bind to internal, or intracellular, receptors that reside within the cell. Many mammalian steroid hormones use this mechanism of cell signaling, as does nitric oxide (NO) gas.
74.7K