Calcium-bound Indo-1
Calcium-bound Indo-1 - Related Videos
08:14
Assessing T-cell Receptor-Induced Calcium Influx Using a Calcium Indicator Dye
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2025
This video analyzes T-cell receptor-induced calcium influx in T-cells. Anti-CD3 antibodies activate the T-cell signaling pathways, leading to an increase in intracellular calcium ions that bind with Indo-1 dye. In flow cytometry, a spectrum shift from the calcium-free state to the calcium-bound state correlates with T-cell receptor-induced calcium influx.
10:42
Flow Cytometric Analysis of Particle-bound Bet v 1 Allergen in PM10
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Cited by 8 •
2016
Here, we present a protocol to quantify allergen-loaded particles by flow cytometry. Ambient particulate matter particles may act as carriers of adsorbed allergens. We show here that flow cytometry, a method widely used to characterize suspended solids >0.5 µm in diameter, can be used to measure these allergen-loaded particles.
03:54
Monitoring B Cell Activation with Antigenic Liposomes using a Calcium-Flux Assay
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2025
This video showcases the assessment of B cell activation through the utilization of antigenic liposomes. B cells treated with calcium-sensitive Indo-1 dye are distinguished using flow cytometry, and their violet-to-blue fluorescence ratio is analyzed. As the cells encounter antigenic liposomes, it triggers B cell activation through calcium influx, progressively enhancing the violet-to-blue fluorescence signal ratio over time.
07:15
Single-Cell Characterization of Calcium Influx and HIV-1 Infection using a Multiparameter Optofluidic Platform
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Cited by 1 •
2021
Here, we present a protocol in which single cells are monitored for acute events and productive HIV-1 infection on a nanofluidic device. Imaging data define virus-host receptor interactions and signaling pathway dynamics. This is the first method for nanofluidic high-throughput longitudinal single-cell culture and imaging to study signaling kinetics and molecular interactions.
06:07
Monitoring PD-1-Blocking Antibodies Bound to T Cells Derived from a Drop of Peripheral Blood
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Cited by 2 •
2020
We developed a simple flow cytometry assay for evaluating the binding of PD-1–blocking antibodies to T cells, requiring only a drop of peripheral blood from cancer patients.
FAQs
The measurement compares emission near 400 nm, associated with the calcium-bound form, with emission near 475 nm, associated with calcium-free Indo-1. As intracellular Ca2+ changes, the balance between these signals changes as well. This ratio provides an estimate of calcium levels while focusing interpretation on the spectral relationship between the two forms.
A ratiometric measurement emphasizes the relative change between two emission signals instead of relying on fluorescence intensity alone. That approach is useful for following rapid calcium fluxes after receptor stimulation or microbial exposure, because the resulting signal can be interpreted as a calcium-associated response rather than simply as a change in overall fluorescence brightness.
Calcium binding changes Indo-1's emission profile, increasing the signal near 400 nm relative to the calcium-free emission near 475 nm. This shift creates the basis for distinguishing calcium-associated and calcium-free indicator states. In immune signaling experiments, the altered emission relationship can therefore be followed over time as intracellular calcium responds to stimulation.
Researchers can monitor the fluorescence ratio during and after a defined receptor stimulus or microbial exposure, then relate its time-dependent changes to intracellular calcium signaling. Rapid increases or other temporal patterns can reveal when cellular activation begins and how signaling develops. The resulting measurements help characterize pathway behavior without treating fluorescence intensity alone as the endpoint.
The approach can be applied to lymphocytes, phagocytes, and other immune cells in which calcium flux contributes to activation or host-pathogen responses. Measuring the ratio after receptor stimulation can support analysis of immune signaling, while exposure to microbes can connect calcium behavior with cellular responses to infection-related challenges.
Indo-1-based calcium measurements can help distinguish cellular signaling responses associated with receptor engagement or microbial exposure. Interpreted alongside the responding cell type and experimental stimulus, the data can characterize activation pathways and functional immune responses. This makes the indicator useful for linking rapid intracellular calcium dynamics with broader immunological or host-pathogen processes.