Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

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,...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

Connectivity Logic of Dendritic Spines in Cortex: Increased Inputs and Ensemble Formation.

bioRxiv : the preprint server for biology·2026

Same author

VesiclePy: A machine learning vesicle analysis toolbox for volume electron microscopy.

PLoS computational biology·2026

Same author

Dense and distributed neuropeptide network in the nerve net of Hydra vulgaris.

PLoS computational biology·2026

Same author

Neuronal ensembles in cortical function and disease.

Physiological reviews·2026

Same author

Ultrastructural reconstruction of the endodermal nerve net of Hydra vulgaris.

Current biology : CB·2025

Same author

Protocol for synthesizing, implanting, and using polydimethylsiloxane as skull replacement in mice for imaging, electrophysiology, and optogenetics.

STAR protocols·2025

Search research articles

Related Experiment Video

Updated: May 10, 2026

Video-rate Scanning Confocal Microscopy and Microendoscopy

Video-rate Scanning Confocal Microscopy and Microendoscopy

Published on: October 20, 2011

How to build a two-photon microscope with a confocal scan head.

Volodymyr Nikolenko, Rafael Yuste

Cold Spring Harbor Protocols

|June 5, 2013

Summary

Convert a standard confocal microscope into a two-photon microscope for cost-effective deep-tissue and live-cell imaging. This DIY approach avoids expensive commercial systems, enabling advanced research without a large budget.

Area of Science:

Biophysics
Microscopy
Neuroscience

Background:

Confocal microscopy is limited in deep-tissue imaging due to scattering and photodamage.
Two-photon microscopy offers superior imaging in scattering tissues and long-term live imaging with reduced photodamage.
Commercial two-photon microscopes are expensive, limiting accessibility for many researchers.

Purpose of the Study:

To provide practical guidelines for converting a standard confocal microscope into a two-photon microscope.
To enable researchers to perform advanced two-photon imaging without significant financial investment.
To facilitate deep-tissue and in vivo imaging of biological samples.

Main Methods:

Detailed instructions for modifying a standard confocal microscope.

More Related Videos

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

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

Published on: February 24, 2026

A Guide to Build a Highly Inclined Swept Tile Microscope for Extended Field-of-view Single-molecule Imaging

A Guide to Build a Highly Inclined Swept Tile Microscope for Extended Field-of-view Single-molecule Imaging

Published on: April 8, 2019

Related Experiment Videos

Last Updated: May 10, 2026

Video-rate Scanning Confocal Microscopy and Microendoscopy

Video-rate Scanning Confocal Microscopy and Microendoscopy

Published on: October 20, 2011

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

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

Published on: February 24, 2026

A Guide to Build a Highly Inclined Swept Tile Microscope for Extended Field-of-view Single-molecule Imaging

A Guide to Build a Highly Inclined Swept Tile Microscope for Extended Field-of-view Single-molecule Imaging

Published on: April 8, 2019

Integration of a two-photon excitation source and associated optical components.

Testing and validation of the converted system for imaging capabilities.

Main Results:

Successful conversion of a confocal microscope to a functional two-photon microscope.
Demonstration of deep-tissue imaging capabilities in scattering preparations.
Validation of long-term live imaging with minimal photodamage.

Conclusions:

DIY two-photon microscope conversion is a feasible and cost-effective alternative to commercial systems.
This approach democratizes access to advanced imaging techniques for biological research.
Enables high-resolution imaging of cells under physiological conditions, including in vivo studies.