Gcamp5 Mcherry
Gcamp5 Mcherry - Related Videos
12:02
Using Tg(Vtg1:mcherry) Zebrafish Embryos to Test the Estrogenic Effects of Endocrine Disrupting Compounds
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Cited by 1 •
2020
Present here is a detailed protocol for the use of zebrafish embryos Tg(vtg1: mCherry) for the detection of estrogenic effects. The protocol covers the propagation of the fish and treatment of embryos, and emphasizes the detection, documentation, and the evaluation of fluorescent signals induced by endocrine disrupting compounds (EDC).
07:04
Imaging Neural Activity in the Primary Somatosensory Cortex Using Thy1-GCaMP6s Transgenic Mice
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Cited by 2 •
2019
We describe an experimental procedure for measuring neuronal activity through dual optical windows above bilateral primary somatosensory corticies (S1) in Thy1-GCaMP6s transgenic mice using 2-photon (2P) microscopy in vivo.
02:31
Ex Vivo Calcium Imaging to Study Drosophila Brain Responses to Neuropeptides
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2025
Source: Ishimoto, H. et. al., Ex Vivo Calcium Imaging for Visualizing Brain Responses to Endocrine Signaling in Drosophila. J. Vis. Exp. (2018) This video demonstrates calcium imaging to study Drosophila brain responses to neuropeptides. Neurons expressing a fluorescent calcium indicator reveal intracellular calcium dynamics and functional responses to signaling molecules.
08:02
Live-cell Imaging of Endocytic Transport using Functionalized Nanobodies in Cultured Cells
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Cited by 1 •
2025
Endocytic and retrograde transport of proteins from the plasma membrane to the trans-Golgi network is essential to maintaining membrane homeostasis and regulating signaling. Here, we describe a method to image and quantify endocytic transport of transmembrane cargo proteins by live-cell microscopy using derivatized anti-GFP nanobodies in HeLa cells.
03:01
Calcium Imaging in Individual Neurons Regulating Egg-Laying Behavior in Caenorhabditis elegans
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2025
This video demonstrates a technique for calcium imaging in individual neurons of behaving Caenorhabditis elegans using genetically encoded fluorescent reporter proteins. It outlines the steps involved in imaging worm movement and measuring reporter fluorescence ratios from neurons to correlate intracellular calcium levels with neuronal activity and behavioral changes during egg-laying.
FAQs
GCaMP5 links calcium binding to a fluorescence change through a conformational mechanism. Its circularly permuted green fluorescent protein is coupled to calmodulin and the M13 peptide; when intracellular calcium binds, these components undergo a conformational change that increases green fluorescence. The signal therefore reports calcium dynamics associated with neuronal activity.
mCherry supplies information that the activity-sensitive green channel cannot provide by itself: a red signal for locating labeled cells or referencing expression. Comparing the two colors helps distinguish where the reporter is present from changes in green fluorescence. This separation supports interpretation of optical recordings across neuronal preparations.
The green signal is most informative when interpreted as an activity-associated calcium signal rather than as a direct label of cell position. Changes in fluorescence can reveal calcium dynamics, whereas the red channel helps identify the labeled population. Together, the channels connect cellular localization with activity-related optical measurements.
An optical recording follows green fluorescence changes while the red fluorescence marks the relevant cells or provides an expression reference. The resulting two-color observation can show which labeled neurons are present and how their calcium-associated signals change. This makes the system suitable for examining activity patterns rather than relying only on cell identification.
At the circuit level, the reporter can help map neural circuits and examine patterns of neuronal communication. In stimulus-response experiments, fluorescence changes provide a way to measure how labeled neurons respond to stimuli. These uses extend the system from single-cell localization to population and circuit-level analyses of activity-associated calcium dynamics.
The system supports optical recordings in cultured neurons, brain tissue, and living animals, allowing researchers to examine calcium dynamics across increasingly complex preparations. The same dual-color logic remains useful in each setting: green fluorescence reports activity-associated calcium changes, while mCherry helps identify labeled cells or provides an expression reference.