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Confocal Fluorescence Microscopy

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,...

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Related Experiment Video

Updated: May 21, 2026

Low-cost Custom Fabrication and Mode-locked Operation of an All-normal-dispersion Femtosecond Fiber Laser for Multiphoton Microscopy

Low-cost Custom Fabrication and Mode-locked Operation of an All-normal-dispersion Femtosecond Fiber Laser for Multiphoton Microscopy

Published on: November 22, 2019

Two-photon microscopy using an Yb(3+)-doped fiber laser with variable pulse widths.

Dong Uk Kim¹, Hoseong Song, Woosub Song

¹School of Information and Mechatronics, Gwangju Institute of Science and Technology, 261 Cheomdan-gwagiro, Buk-gu, Gwangju 500-712, South Korea.

|June 21, 2012

Summary

This study presents a novel confocal two-photon microscope system utilizing a ytterbium-doped fiber laser, optimizing pulse width for enhanced two-photon excitation. The system successfully imaged neural cells, demonstrating its effectiveness for biological research.

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20 mJ, 1 ps Yb:YAG Thin-disk Regenerative Amplifier

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Last Updated: May 21, 2026

Low-cost Custom Fabrication and Mode-locked Operation of an All-normal-dispersion Femtosecond Fiber Laser for Multiphoton Microscopy

Low-cost Custom Fabrication and Mode-locked Operation of an All-normal-dispersion Femtosecond Fiber Laser for Multiphoton Microscopy

Published on: November 22, 2019

20 mJ, 1 ps Yb:YAG Thin-disk Regenerative Amplifier

20 mJ, 1 ps Yb:YAG Thin-disk Regenerative Amplifier

Published on: July 12, 2017

Area of Science:

Biophysics
Optical Microscopy
Laser Technology

Background:

Two-photon fluorescence microscopy commonly employs femtosecond Ti:Sapphire lasers near 800 nm.
Limitations exist in current laser sources for optimal two-photon excitation.

Purpose of the Study:

Introduce a new confocal two-photon microscope system.
Utilize a mode-locked ytterbium (Yb3+)-doped fiber laser for improved performance.
Optimize laser pulse characteristics for enhanced two-photon excitation.

Main Methods:

Employed a mode-locked Yb3+-doped fiber laser generating 13 ps pulses.
Used an external grating pair pulse compressor to control pulse width and frequency chirping.
Optimized two-photon excitation by monitoring photocurrent in a GaAsP photodiode.

Main Results:

Achieved controlled pulse width and frequency chirping for optimal two-photon excitation.
Demonstrated effective imaging of Vybrant DiI-stained dorsal root ganglion cells.
Successfully performed 2D and 3D imaging with the optimized system.

Conclusions:

The developed confocal two-photon microscope system offers a viable alternative to traditional Ti:Sapphire lasers.
Pulse width optimization is crucial for efficient two-photon excitation and high-quality imaging.
The system shows promise for advanced biological imaging applications.