Intracellular Ph Imaging
Intracellular Ph Imaging - Related Videos
07:55
Imaging the Intracellular Trafficking of APP with Photoactivatable GFP
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Cited by 6 •
2015
While the transport of cell surface proteins is relatively easily studied, visualizing the trafficking of intracellular proteins is much more difficult. Here, we use constructs incorporating photoactivatable GFP and demonstrate a method to accurately follow the amyloid precursor protein from the Golgi apparatus to down-stream compartments and follow its clearance.
03:43
High-Throughput Imaging of Intracellular Brucella abortus Infection in Host Epithelial Cells
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2026
Source: Casanova, A., et al. Microscopy-based Assays for High-throughput Screening of Host Factors Involved in Brucella Infection of Hela Cells. J. Vis. Exp. (2016)This video demonstrates high-throughput imaging of Brucella abortus infection in epithelial cells with siRNA-mediated STING knockdown to investigate the role of cytosolic DNA sensing in intracellular bacterial replication and host innate immune response.
12:58
Visualisation and Quantification of Intracellular Interactions of Neisseria meningitidis and Human α-actinin by Confocal Imaging
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2010
Neisseria meningitidis (Nm), a gram negative human-specific respiratory pathogen, can bind to human α-actinin. Here we present a protocol for visualisation of colocalisation of the bacterium with intracellular α-actinin after bacterial entry into human brain microvascular endothelial cells (HBMECs).
07:18
Intracellular Refolding Assay
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Cited by 9 •
2012
In this protocol a method to measure intracellular protein refolding after heat shock is described. This method can be used to study foldases like molecular chaperones and their co-factors or compounds able to influence their activity. Firefly luciferase activity is used as reporter to measure chaperone refolding activity.
03:40
Imaging Intracellular ATP Dynamics Using FRET-Based Sensors in an Organotypic Mouse Hippocampal Slice
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2025
Source: Lerchundi, R., et. al. Imaging of Intracellular ATP in Organotypic Tissue Slices of the Mouse Brain using the FRET-based Sensor ATeam1.03YEMK. J. Vis. Exp. (2019)This video demonstrates the use of FRET(Fluorescence Resonance Energy Transfer)-based sensors to measure intracellular ATP levels in organotypic mouse hippocampal slices, enabling real-time visualization of ATP dynamics in neurons and astrocytes under physiological and experimental conditions.
FAQs
These probes alter their fluorescence intensity or emission properties when proton concentration changes. The resulting optical signal provides a readout that can be tracked within individual cells and across subcellular regions. Because the signal responds to acidity or alkalinity, changes over time can be related to cellular processes that modify intracellular chemical conditions.
Calibration links a probe’s optical response to intracellular pH values rather than leaving results as unscaled fluorescence changes. Ratiometric measurements use changes in optical properties to improve interpretation of proton concentration differences. Together, these approaches help researchers compare measurements between cells, conditions, or experiments and distinguish pH changes from simple signal variation.
Imaging can distinguish pH conditions in the cytosol from those in cellular organelles, allowing researchers to examine how acidity varies across compartments. This spatial information complements measurements of overall cell behavior. Comparing compartments can show whether a treatment or cellular event produces a localized pH disturbance or a broader change throughout the cell.
A typical workflow applies a pH-sensitive fluorescent probe to living cells, records its intensity or emission response, and relates the optical data to pH through calibration or ratiometric analysis. Researchers then examine the measurements across cellular locations or time points. This sequence converts fluorescence observations into interpretable intracellular pH information.
The technique is useful when researchers need to follow pH changes during cellular processes rather than measure only a final endpoint. It can monitor dynamic shifts while connecting molecular events with whole-cell physiology. Spatial and temporal measurements also support comparisons between cytosolic and organelle environments, helping identify where and when pH regulation changes.
Researchers can compare intracellular pH patterns in untreated and treated cells or examine differences associated with disease-related disruption. Changes in fluorescence-derived pH values may indicate altered metabolism, signaling, transport, or other cell functions influenced by acidity and alkalinity. The measurements therefore provide a way to connect experimental conditions with physiological consequences at cellular and subcellular scales.