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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 16, 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

Fast STED microscopy with continuous wave fiber lasers.

Gael Moneron¹, Rebecca Medda, Birka Hein

¹Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany.

|February 23, 2010

Summary

This summary is machine-generated.

We demonstrate fast, continuous-wave stimulated-emission-depletion (CW-STED) microscopy using affordable fiber lasers. This technique achieves high resolution for imaging cellular structures and dynamics in living cells.

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Last Updated: Jun 16, 2026

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

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Published on: November 22, 2019

High-Throughput Total Internal Reflection Fluorescence and Direct Stochastic Optical Reconstruction Microscopy Using a Photonic Chip

High-Throughput Total Internal Reflection Fluorescence and Direct Stochastic Optical Reconstruction Microscopy Using a Photonic Chip

Published on: November 16, 2019

Area of Science:

Optics and Photonics
Biomedical Imaging
Cell Biology

Background:

Stimulated-emission-depletion (STED) microscopy offers super-resolution imaging but often relies on complex and expensive laser systems.
Continuous-wave (CW) lasers are desirable for STED due to their stability and cost-effectiveness, but achieving high resolution with CW-STED can be challenging.

Purpose of the Study:

To develop and demonstrate a fast beam-scanning STED microscopy system in the visible range.
To achieve high spatial resolution using compact, low-cost, turn-key CW fiber lasers.
To visualize dynamic cellular processes in living cells with enhanced resolution.

Main Methods:

Utilized a compact, turn-key continuous wave (CW) fiber laser emitting at 592 nm for STED excitation and depletion.
Implemented a fast beam-scanning approach for rapid image acquisition.
Applied the system to image various samples, including fluorescent nanoparticles, immuno-stained cells with organic fluorescent markers, and living cells expressing yellow fluorescent protein (Citrine).

Main Results:

Achieved spatial resolutions ranging from 35 to 65 nm in the focal plane across different samples.
Demonstrated the imaging of fluorescent nanoparticles, fixed cells with common organic markers, and living cells.
Acquired a movie of the endoplasmic reticulum (ER) in a living cell (6 microm x 12 microm) with 100 frames, each captured in less than 0.2 seconds.

Conclusions:

The combination of CW-STED microscopy and fast beam scanning provides a powerful and accessible method for super-resolution imaging.
This approach enables high-resolution visualization of cellular structures and dynamics in living systems with improved speed and reduced cost.
The system shows significant potential for routine biological research requiring fast, high-resolution live-cell imaging.