Lck-rcamp2
Lck-rcamp2 - Related Videos
02:33
Spontaneous Calcium Imaging for Compartment-Specific Dynamics in Astrocytes
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2025
Source: Bannai, H., et. al., Dissection of Local Ca2+ Signals in Cultured Cells by Membrane-targeted Ca2+ Indicators. J. Vis. Exp. (2019)The video demonstrates fluorescence imaging to observe compartment-specific spontaneous calcium dynamics in astrocytes by sequentially monitoring calcium-sensitive proteins localized on the plasma membrane and endoplasmic reticulum.
11:45
Imaging of HIV-1 Envelope-induced Virological Synapse and Signaling on Synthetic Lipid Bilayers
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Cited by 4 •
2012
This article describes a method to visualize formation of an HIV-1 envelope-induced virological synapse on glass supported planar bilayers by total internal reflection fluorescence (TIRF) microscopy. The method can also be combined with immunofluorescence staining to detect activation and redistribution of signaling molecules that occur during HIV-1 envelope-induced virological synapse formation.
07:42
In-vivo Detection of Protein-protein Interactions on Micro-patterned Surfaces
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Cited by 7 •
2010
This video shows experiments with subsequent analysis of protein-protein interactions by the use of micro-patterned surfaces. The approach offers the possibility to detect protein interactions in living cells and combines high throughput capabilities with the possibility to extract quantitative information.
09:24
Generation of Microtumors Using 3D Human Biogel Culture System and Patient-derived Glioblastoma Cells for Kinomic Profiling and Drug Response Testing
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Cited by 20 •
2016
Patient-derived xenografts of glioblastoma multiforme can be miniaturized into living microtumors using 3D human biogel culture system. This in vivo-like 3D tumor assay is suitable for drug response testing and molecular profiling, including kinomic analysis.
16:10
A TIRF Microscopy Technique for Real-time, Simultaneous Imaging of the TCR and its Associated Signaling Proteins
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Cited by 13 •
2012
The compartmentalization of proteins either within the plasma membrane or into intracellular locations is one regulatory mechanism that can greatly influence signaling outcomes; hence, to understand signaling it is important to study the spatial and temporal behavior of the proteins involved. We describe here a TIRF microscopy based system to study signal transduction in T cells, but is broadly applicable.
FAQs
The Lck-derived membrane-targeting sequence positions the calcium sensor close to the plasma membrane rather than distributing the signal uniformly throughout the cell. This placement increases sensitivity to calcium events occurring at the cell surface and helps distinguish localized activity from broader intracellular changes. In neurons, that spatial bias is valuable for examining signaling in dendrites, axons, and synaptic regions.
Calcium binding changes the conformation of the rCaMP2 sensor, which increases its red fluorescence intensity. The fluorescence response therefore serves as an optical indicator of local calcium elevation rather than a direct measurement of electrical activity. Because the sensor is membrane-positioned, changes in brightness can be interpreted in relation to calcium entry near the neuronal plasma membrane.
Membrane localization links the optical signal to calcium dynamics near sites where neuronal signaling is organized. This can help separate spatially restricted influx from calcium changes occurring elsewhere in the cell. The resulting resolution supports investigations of how localized signals are arranged within dendrites, axons, and synaptic regions, where the location of calcium activity may be as informative as its presence.
A study can use neurons expressing the genetically encoded indicator and record red fluorescence while neuronal activity occurs. Increases in fluorescence indicate calcium-dependent sensor responses near the plasma membrane. Imaging selected cellular regions then allows researchers to examine where activity-associated calcium signals arise, including dendritic, axonal, or synaptic compartments in living cells or neural circuits.
Lck-rCaMP2 is suited to measurements in dendrites, axons, and synaptic regions because its fluorescence is concentrated near the cell surface. Comparing signals across these compartments can reveal whether calcium activity is spatially restricted or distributed across a neuron. This makes the indicator relevant to studies of compartment-specific signaling rather than only whole-cell calcium changes.
The indicator can support research on neuronal excitability, synaptic transmission, and the subcellular organization of calcium signaling. Its red fluorescence provides a way to follow activity-related calcium dynamics in living cells and neural circuits, while membrane targeting preserves information about signal location. These features help connect calcium influx with the specific neuronal structures where signaling occurs.