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

Photoreceptors and Visual Pathways

Photoreceptors and Visual Pathways

At the molecular level, visual signals trigger transformations in photopigment molecules, resulting in changes in the photoreceptor cell's membrane potential. The photon's energy level is denoted by its wavelength, with each specific wavelength of visible light associated with a distinct color. The spectral range of visible light, classified as electromagnetic radiation, spans from 380 to 720 nm. Electromagnetic radiation wavelengths exceeding 720 nm fall under the infrared category,...

Fluorescence and Phosphorescence: Instrumentation

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.

You might also read

Related Articles

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

Sort by

Same author

Voltage dynamics of cortical dendrites in vivo.

Nature neuroscience·2026

Same author

Correction to "Optical Single-Channel Recording via Diffusional Confinement in Membrane Tethers".

ACS nano·2026

Same author

<math><mrow><mi>Micro</mi> <mi>S</mi> <mi>plit</mi></mrow></math> : semantic unmixing of fluorescent microscopy data.

Nature methods·2026

Same author

A dendrite-resolved, <i>in vivo</i> transfer function from spike patterns to dendritic Ca<sup>2</sup>.

bioRxiv : the preprint server for biology·2026

Same author

Ether lipids influence cancer cell fate by modulating iron uptake.

Nature communications·2026

Same author

Fast dendritic excitations primarily mediate back-propagation in CA1 pyramidal neurons during behavior.

bioRxiv : the preprint server for biology·2026

Same journal

Taphonomic analysis at Liang Bua reveals the behavioral and technological capabilities of <i>Homo floresiensis</i>.

Science advances·2026

Same journal

Targeting granule initiation and amyloplast structure to create giant starch granules in wheat.

Science advances·2026

Same journal

A meta-analysis of carbon losses and gains from tropical moist forest degradation and regeneration.

Science advances·2026

Same journal

Ancient DNA reveals elite dynastic rule among Iron Age Eurasian Steppe nomads.

Science advances·2026

Same journal

Targeting astrocytic Dp71 attenuates BBB disruption after traumatic brain injury through WTAP-associated m<sup>6</sup>A regulation of MMP2.

Science advances·2026

Same journal

Pancreatic α cells are required for nutrient homeostasis by regulating dynamic β cell networks in islets.

Science advances·2026

See all related articles

Search research articles

Related Experiment Video

Updated: Jun 3, 2025

Imaging Membrane Potential with Two Types of Genetically Encoded Fluorescent Voltage Sensors

Imaging Membrane Potential with Two Types of Genetically Encoded Fluorescent Voltage Sensors

Published on: February 4, 2016

Photophysics-informed two-photon voltage imaging using FRET-opsin voltage indicators.

F Phil Brooks¹, Daozheng Gong¹, Hunter C Davis¹

¹Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, USA.

Science Advances

|January 8, 2025

Summary

This summary is machine-generated.

Genetically encoded voltage indicators (GEVIs) show voltage sensitivity issues under two-photon excitation. Optimized protocols enable high-speed in vivo voltage imaging using FRET-opsin GEVIs like Voltron2.

More Related Videos

Single-Cell Optical Action Potential Measurement in Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes

Single-Cell Optical Action Potential Measurement in Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes

Published on: December 22, 2020

In vivo Quantification of G Protein Coupled Receptor Interactions using Spectrally Resolved Two-photon Microscopy

In vivo Quantification of G Protein Coupled Receptor Interactions using Spectrally Resolved Two-photon Microscopy

Published on: January 19, 2011

Related Experiment Videos

Last Updated: Jun 3, 2025

Imaging Membrane Potential with Two Types of Genetically Encoded Fluorescent Voltage Sensors

Imaging Membrane Potential with Two Types of Genetically Encoded Fluorescent Voltage Sensors

Published on: February 4, 2016

Single-Cell Optical Action Potential Measurement in Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes

Single-Cell Optical Action Potential Measurement in Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes

Published on: December 22, 2020

In vivo Quantification of G Protein Coupled Receptor Interactions using Spectrally Resolved Two-photon Microscopy

In vivo Quantification of G Protein Coupled Receptor Interactions using Spectrally Resolved Two-photon Microscopy

Published on: January 19, 2011

Area of Science:

Neuroscience
Biophysics
Molecular Biology

Background:

Microbial rhodopsin-derived genetically encoded voltage indicators (GEVIs) are crucial for mapping bioelectrical activity.
Förster resonance energy transfer (FRET)-opsin GEVIs offer high brightness, speed, and voltage sensitivity.
A significant limitation is the reduced voltage sensitivity of FRET-opsin GEVIs under two-photon (2P) excitation.

Purpose of the Study:

To investigate the photophysics of FRET-opsin GEVIs Voltron1 and Voltron2.
To understand the reasons behind decreased voltage sensitivity under 2P excitation.
To develop optimized protocols for effective 2P voltage imaging in vivo.

Main Methods:

Photophysical characterization of Voltron1 and Voltron2.
Analysis of voltage sensitivity as a function of illumination intensity.
Development and application of photocycle-optimized 2P illumination protocols.
In vivo 2P voltage imaging in a live mouse model.

Main Results:

The observed voltage sensitivity in FRET-opsin GEVIs originates from photocycle intermediates, not ground states.
Voltage sensitivity is a nonlinear function of illumination intensity, with sign reversal possible under low intensity.
Photocycle-optimized 2P illumination protocols were successfully developed.
High-speed 2P voltage imaging was demonstrated in vivo using Voltron2 in mouse brain.

Conclusions:

The photophysics of FRET-opsin GEVIs under 2P excitation are complex and influenced by photocycle intermediates and illumination intensity.
Optimized 2P illumination protocols can overcome previous limitations, enabling effective in vivo voltage imaging.
These findings pave the way for advanced high-speed 2P voltage imaging applications in neuroscience.