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Related Concept Videos

Super-resolution Fluorescence Microscopy01:37

Super-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.
Two-Dimensional Microscopy in Microbiology01:29

Two-Dimensional Microscopy in Microbiology

Two-dimensional (2D) microscopy encompasses a range of optical techniques that capture images within a single focal plane, offering detailed representations of microscopic structures. These techniques are essential in biological and medical research, enabling the visualization of cellular and subcellular structures with different levels of contrast and specificity.There are several major types of 2D microscopy, each with strengths and applications.Bright-Field MicroscopyBright-field microscopy...
Confocal Fluorescence Microscopy01:16

Confocal 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,...
Immunofluorescence Microscopy01:12

Immunofluorescence Microscopy

A fluorescence microscope uses fluorescent chromophores called fluorochromes, which can absorb energy from a light source and then emit this energy as visible light. Fluorochromes include naturally fluorescent substances (such as chlorophylls) and fluorescent stains that are added to the specimen to create contrast. Dyes such as Texas red and FITC are examples of fluorochromes. Other examples include the nucleic acid dyes 4’,6’-diamidino-2-phenylindole (DAPI), and acridine orange.
The...
Total Internal Reflection Fluorescence Microscopy01:05

Total Internal Reflection Fluorescence Microscopy

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.
Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

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...

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Conducting Multiple Imaging Modes with One Fluorescence Microscope
08:32

Conducting Multiple Imaging Modes with One Fluorescence Microscope

Published on: October 28, 2018

Fluorescence microscopy.

Sébastien Peter1, Klaus Harter, Frank Schleifenbaum

  • 1Center for Plant Molecular Biology, University of Tuebingen, Tuebingen, Germany.

Methods in Molecular Biology (Clifton, N.J.)
|September 24, 2013
PubMed
Summary
This summary is machine-generated.

Optical microscopy is crucial for Arabidopsis cell biology, offering high sensitivity and resolution. Spectroscopic techniques provide functional insights beyond simple intensity measurements for cellular processes.

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Last Updated: May 7, 2026

Conducting Multiple Imaging Modes with One Fluorescence Microscope
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Area of Science:

  • Plant cell biology
  • Biophysics
  • Molecular imaging

Background:

  • Optical microscopy is essential for studying plant cells.
  • Autofluorescent proteins enable specific protein tracking.
  • Quantitative and functional analysis requires advanced techniques.

Purpose of the Study:

  • To explain the theory of fluorescence emission for optical microscopy.
  • To detail spectroscopic techniques for functional cell biology.
  • To integrate intensity-based and spectroscopic methods.

Main Methods:

  • Theoretical explanation of fluorescence emission.
  • Focus on spectroscopic techniques.
  • Application to quantitative cell biology.

Main Results:

  • Provides theoretical foundation for fluorescence microscopy.
  • Highlights spectroscopic methods for functional insights.
  • Enables quantitative and functional analysis of cellular processes.

Conclusions:

  • Optical microscopy, especially with spectroscopy, is vital for Arabidopsis cell biology.
  • Advanced techniques enhance understanding of cellular functions.
  • This chapter provides a theoretical basis for practical applications.