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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: Jun 24, 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

Highly versatile confocal microscopy system based on a tunable femtosecond Er:fiber source.

D Träutlein¹, F Adler, K Moutzouris

¹Department of Biology and Center for Applied Photonics, University of Konstanz, 78457 Konstanz, Germany.

Journal of Biophotonics

|April 4, 2009

Summary

This study demonstrates a tunable femtosecond fiber laser for advanced microscopy. It enables efficient linear and nonlinear imaging, including two-photon microscopy for deep-tissue applications and three-photon absorption for DNA research.

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Video-rate Scanning Confocal Microscopy and Microendoscopy

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Published on: October 20, 2011

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Last Updated: Jun 24, 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

Video-rate Scanning Confocal Microscopy and Microendoscopy

Video-rate Scanning Confocal Microscopy and Microendoscopy

Published on: October 20, 2011

Area of Science:

Optics and Photonics
Biomedical Imaging
Laser Physics

Background:

Confocal microscopy requires optimized light sources for high-resolution imaging.
Linear and nonlinear imaging techniques offer complementary information but demand versatile laser systems.

Purpose of the Study:

To evaluate a single femtosecond fiber laser system for both linear and nonlinear microscopy.
To explore the system's performance across a wide range of pump wavelengths.
To demonstrate its capabilities for advanced imaging applications.

Main Methods:

Utilized a tunable femtosecond fiber laser system.
Employed linear fluorescence excitation and nonlinear imaging techniques (two-photon and three-photon absorption).
Optimized pulse duration using a prism compressor for dispersion control.

Main Results:

Demonstrated benefits of a tunable visible spectrum laser for fluorescence excitation.
Achieved sub-33 fs pulse durations in the confocal region.
Enabled two-photon microscopy of mouse brain tissue with 2 mW average power at 1100 nm.
Induced localized photodamage in DNA using three-photon absorption.

Conclusions:

The compact femtosecond fiber laser system is highly versatile for diverse microscopy applications.
The system shows significant potential for deep-tissue imaging in the near-infrared window.
Higher-order nonlinearities can be effectively exploited for precise biological sample manipulation.