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Related Concept Videos

Super-resolution Fluorescence Microscopy01:37

Super-resolution Fluorescence Microscopy

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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...
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Confocal Fluorescence Microscopy01:16

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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,...
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Two-Dimensional Microscopy in Microbiology01:29

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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...
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Photoluminescence offers a wide range of applications due to its inherent sensitivity and selectivity. This technique allows for both direct and indirect analyses of the analyte. Direct quantitative analysis is possible when the analyte exhibits a favorable quantum yield for fluorescence or phosphorescence. However, an indirect analysis may be feasible if the analyte is not fluorescent or phosphorescent, or if the quantum yield is unfavorable. Indirect methods include reacting the analyte with...
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Total Internal Reflection Fluorescence Microscopy01:05

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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.
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Immunofluorescence Microscopy01:12

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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.
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Related Experiment Video

Updated: May 4, 2026

Conducting Multiple Imaging Modes with One Fluorescence Microscope
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Fluorescence nanoscopy. Methods and applications.

Jose Requejo-Isidro1

  • 1Unidad de Biofísica, CSIC-UPV/EHU, Barrio de Sarriena, s/n., 48940 Leioa, Spain.

Journal of Chemical Biology
|January 17, 2014
PubMed
Summary
This summary is machine-generated.

This review covers fluorescence nanoscopy, a technique achieving super-resolution imaging beyond conventional limits. It explores advanced acquisition, labeling, and quantitative methods for detailed biological insights.

Keywords:
Fluorescence nanoscopyPALMSIMSTEDSTORMSubdiffraction imagingSuper-resolution

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Area of Science:

  • Biophysics
  • Optical Microscopy
  • Nanotechnology

Background:

  • Conventional fluorescence microscopy is limited by diffraction, hindering detailed visualization of cellular structures.
  • Fluorescence nanoscopy overcomes these limitations, enabling imaging at the nanoscale.

Purpose of the Study:

  • To explain the fundamental concepts of fluorescence nanoscopy.
  • To highlight recent advancements in acquisition techniques and labeling strategies.
  • To discuss quantitative methods for analyzing super-resolved images.

Main Methods:

  • Review of established and emerging fluorescence nanoscopy techniques.
  • Analysis of novel fluorescent probes and labeling approaches.
  • Examination of computational and statistical methods for data analysis.

Main Results:

  • Detailed explanation of various super-resolution microscopy principles.
  • Overview of optimized labeling strategies for enhanced contrast and specificity.
  • Introduction to quantitative analysis tools for extracting biological information.

Conclusions:

  • Fluorescence nanoscopy provides unprecedented resolution for biological imaging.
  • Continued innovation in techniques and analysis promises deeper understanding of cellular processes.