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Related Concept Videos

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,...
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.
Fluorescence and Phosphorescence: Instrumentation01:25

Fluorescence and Phosphorescence: Instrumentation

Fluorometers and spectrofluorometers are two types of instruments used for measuring molecular fluorescence. These instruments differ in how they select excitation and emission wavelengths and the type of light sources they utilize. Fluorometers use absorption interference filters to choose excitation and emission wavelengths. The excitation source in a fluorometer is typically a low-pressure mercury vapor lamp that emits intense lines distributed throughout the ultraviolet and visible regions.
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.
Phase Contrast and Differential Interference Contrast Microscopy01:26

Phase Contrast and Differential Interference Contrast Microscopy

Phase-Contrast Microscopes
In-phase-contrast microscopes, interference between light directly passing through a cell and light refracted by cellular components is used to create high-contrast, high-resolution images without staining. It is the oldest and simplest type of microscope that creates an image by altering the wavelengths of light rays passing through the specimen. Altered wavelength paths are created using an annular stop in the condenser. The annular stop produces a hollow cone of...
Imaging Biological Samples with Optical Microscopy01:18

Imaging Biological Samples with Optical Microscopy

Optical microscopy uses optic principles to provide detailed images of samples. Antonie van Leeuwenhoek designed the first compound optical microscope in the 17th century to visualize blood cells, bacteria, and yeast cells. In 1830, Joseph Jackson Lister created an essentially modern light microscope. The 20th century saw the development of microscopes with enhanced magnification and resolution.
In optical microscopy, the specimen to be viewed is placed on a glass slide and clipped on the stage...

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Updated: May 10, 2026

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy
12:51

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy

Published on: December 9, 2013

Coherent Control in Multiphoton Fluorescence Imaging.

Arijit Kumar De1, Debabrata Goswami

  • 1Deptartment of Chemistry, Indian Institute of Technology Kanpur, UP - 208016, India.

Proceedings of Spie--The International Society for Optical Engineering
|July 2, 2013
PubMed
Summary
This summary is machine-generated.

Ultrafast laser pulses enhance multiphoton fluorescence microscopy by controlling pulse properties. Optimizing pulse timing and shaping improves signal and allows selective fluorophore excitation for advanced imaging applications.

Keywords:
Coherent controlamplitude modulationmultiphoton imagingpulse shapingpump-probe spectroscopytime-domain controlultrafast imagingultrafast laser pulses

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Super-Resolution Imaging and Shared Management: A Protocol for Confocal Microscopy with Multiplex Detection
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Super-Resolution Imaging and Shared Management: A Protocol for Confocal Microscopy with Multiplex Detection

Published on: February 24, 2026

Related Experiment Videos

Last Updated: May 10, 2026

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy
12:51

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy

Published on: December 9, 2013

Super-Resolution Imaging and Shared Management: A Protocol for Confocal Microscopy with Multiplex Detection
07:42

Super-Resolution Imaging and Shared Management: A Protocol for Confocal Microscopy with Multiplex Detection

Published on: February 24, 2026

Area of Science:

  • Optics and Photonics
  • Biomedical Imaging
  • Laser Physics

Background:

  • Multiphoton fluorescence microscopy utilizes ultrafast laser pulses to overcome low fluorophore absorption cross-sections.
  • Standard techniques often face limitations in signal enhancement and specificity.

Purpose of the Study:

  • To explore methods for enhancing signal generation in multiphoton microscopy.
  • To investigate the impact of laser pulse manipulation on imaging fidelity and fluorophore selectivity.

Main Methods:

  • Amplitude modulation of ultrashort pulse trains to enhance two-photon fluorescence.
  • Control of time lag between phase-locked laser pulses for imaging optimization.
  • Laser pulse-shaping techniques for temporal pulse compression and frequency component manipulation.

Main Results:

  • Amplitude modulation provides insight into laser-induced photo-thermal damage.
  • Controlling time lag between pulses affects imaging outcomes.
  • Pulse-shaping offers potential for signal enhancement and selective fluorophore excitation.

Conclusions:

  • Advanced control over ultrafast laser pulse parameters significantly enhances multiphoton fluorescence microscopy.
  • Pulse-shaping techniques hold promise for future applications in selective bioimaging and diagnostics.