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

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

Beyond transport: Lysosomal solute carriers as orchestrators of immune signaling.

Molecular biology of the cell·2026

Same author

Coupling dead cell recognition to Fcγ receptors augments anticancer immunity.

Nature cancer·2026

Same author

Lipid scrambling via TMEM16F mediates the formation and release of extracellular vesicles.

Molecular biology of the cell·2026

Same author

Multimodal antigenic escape to GPRC5D-targeted T cell engagers in multiple myeloma.

Nature medicine·2026

Same author

Is the Parkinson's-associated protein TMEM175 a proton channel: Yay or nay?

The Journal of cell biology·2025

Same author

DNGR-1 signalling limits dendritic cell activation for optimal antigen cross-presentation.

The EMBO journal·2025

Same journal

Mapping the 3D Chromosome Organization of a Biosynthetic Gene Cluster by Capture Hi-C (CHi-C).

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

Mapping the 3D Chromosome Organization of Streptomyces by Hi-C.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

CUT&Tag Epigenomic Profiling of Biosynthetic Gene Clusters in Arabidopsis thaliana.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

Rhizobium rhizogenes-Mediated Hairy Root Transformation Protocol for Lotus japonicus and Other Legumes.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

Characterization of Bioactive Saponins from Sea Cucumbers.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

Methods for Functional Validation of Terpenoid Metabolic Clusters in Nicotiana benthamiana and Aspergillus oryzae.

Methods in molecular biology (Clifton, N.J.)·2026

See all related articles

Search research articles

Related Experiment Video

Updated: Mar 12, 2026

Measuring Phagosome pH by Ratiometric Fluorescence Microscopy

Measuring Phagosome pH by Ratiometric Fluorescence Microscopy

Published on: December 7, 2015

Measuring Phagosomal pH by Fluorescence Microscopy.

Johnathan Canton¹, Sergio Grinstein^2,3

¹Program in Cell Biology, Peter Gilgan Centre for Research and Learning, Hospital for Sick Children, 686 Bay Street, 19-9800, Toronto, ON, Canada, M5G 0A4.

Methods in Molecular Biology (Clifton, N.J.)

|November 6, 2016

Summary

This summary is machine-generated.

Dual wavelength ratiometric imaging offers dynamic pH monitoring in live cells, overcoming limitations of other methods. This technique allows for precise measurements within individual cells and organelles, enhancing intracellular studies.

Keywords:

Dual wavelength ratio imaging Fluorescein Fluorescence microscopy Oregon Green Phagosome SNARF1 pH

More Related Videos

Imaging the Neutrophil Phagosome and Cytoplasm Using a Ratiometric pH Indicator

Imaging the Neutrophil Phagosome and Cytoplasm Using a Ratiometric pH Indicator

Published on: April 5, 2017

Study of Phagolysosome Biogenesis in Live Macrophages

Study of Phagolysosome Biogenesis in Live Macrophages

Published on: March 10, 2014

Related Experiment Videos

Last Updated: Mar 12, 2026

Measuring Phagosome pH by Ratiometric Fluorescence Microscopy

Measuring Phagosome pH by Ratiometric Fluorescence Microscopy

Published on: December 7, 2015

Imaging the Neutrophil Phagosome and Cytoplasm Using a Ratiometric pH Indicator

Imaging the Neutrophil Phagosome and Cytoplasm Using a Ratiometric pH Indicator

Published on: April 5, 2017

Study of Phagolysosome Biogenesis in Live Macrophages

Study of Phagolysosome Biogenesis in Live Macrophages

Published on: March 10, 2014

Area of Science:

Cell biology
Microscopy
Biochemistry

Background:

Intracellular pH dynamics are crucial for cellular processes.
Traditional methods struggle with live-cell imaging and resolving individual organelles.
Dual wavelength ratiometric imaging provides a solution for dynamic pH studies.

Purpose of the Study:

To discuss the principles of dual wavelength ratiometric imaging.
To detail its application in measuring phagosomal pH.
To guide probe selection, instrumentation, and calibration.

Main Methods:

Utilizing dual wavelength ratiometric imaging for live-cell microscopy.
Accounting for focal plane variations, probe loading, and photobleaching.
Applying the technique to measure phagosomal pH.

Main Results:

Ratiometric imaging enables dynamic monitoring of intracellular pH.
It resolves individual cells and organelles, unlike population methods.
The method accounts for common imaging artifacts.

Conclusions:

Dual wavelength ratiometric imaging is a powerful tool for studying intracellular pH.
It offers advantages over whole population methods for high-resolution studies.
The chapter provides a comprehensive guide to its application in phagosomal pH measurement.