H2b-mcherry Labeling
H2b-mcherry Labeling - 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).
09:19
Simultaneous Label-Free Autofluorescence Multi-Harmonic Microscopy
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Cited by 3 •
2025
This protocol presents a step-by-step guide for the Simultaneous Label-free Autofluorescence Multi-harmonic (SLAM) microscopic technique, including details on how to generate the laser light source, prepare a tissue sample, conduct imaging, and analyze the data. SLAM advances nonlinear microscopy by measuring four complementary label-free contrasts to investigate the tissue microenvironment.
08:28
Metabolic Labeling
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2023
Metabolic labeling is used to probe the biochemical transformations and modifications that occur in a cell. This is accomplished by using chemical analogs that mimic the structure of natural biomolecules. Cells utilize analogs in their endogenous biochemical processes, producing compounds that are labeled. The label allows for the incorporation of detection and affinity tags, which can then be used to elucidate metabolic pathways using other biochemical analytical techniques, such as SDS-PAGE...
10:13
Measuring G-protein-coupled Receptor Signaling via Radio-labeled GTP Binding
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Cited by 11 •
2017
Guanosine triphosphate (GTP) binding is one of the earliest events in G-Protein-Coupled Receptor (GPCR) activation. This protocol describes how to pharmacologically characterize specific GPCR-ligand interactions by monitoring the binding of the radio-labeled GTP analog, [35S]guanosine-5'-O-(3-thio)triphosphate ([35S]GTPγS), in response to a ligand of interest.
09:41
Visualization of Cell Cycle Variations and Determination of Nucleation in Postnatal Cardiomyocytes
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Cited by 5 •
2017
To distinguish cell division from cell cycle variations in cardiomyocytes, we present protocols using two transgenic mouse lines: Myh6-H2B-mCh transgenic mice, for the unequivocal identification of cardiomyocyte nuclei, and CAG-eGFP-anillin mice, for distinguishing cell division from cell cycle variations.
FAQs
Histone H2B is a core protein of nucleosomes, the structures that organize DNA. When H2B is genetically fused to mCherry, the fluorescent component follows H2B as it becomes incorporated into nucleosomes. This produces a chromatin-associated signal distributed through the nucleus, allowing nuclear organization to be observed without relying on a separate, nonlocalized fluorescent marker.
Chromatin incorporation links the fluorescent signal to the material that packages nuclear DNA. As a result, changes in the signal’s spatial pattern can be interpreted in relation to nuclear shape and organization rather than fluorescence alone. This relationship is especially useful when following nuclei through cellular behaviors such as division, migration, or changes in tissue organization.
The labeling provides a persistent nuclear reference that supports time-lapse observation of individual cells. Researchers can use changes in nuclear position, shape, and number to follow cell migration, identify cell division, and track lineage behavior over time. Because the signal preserves spatial information, these observations can be related to neighboring cells and the surrounding engineered tissue or culture.
In living cells, the fluorescent nuclear signal enables repeated imaging of the same cells across successive time points. This makes it possible to connect earlier positions or nuclear morphologies with later outcomes, rather than analyzing separate snapshots. Such longitudinal observation helps reveal dynamic responses and cell-to-cell behavior in engineered tissues or cell cultures.
A study first uses the genetic H2B-mCherry fusion so the fluorescent tag can associate with nucleosomes. Cells or engineered tissues are then observed under conditions suitable for fluorescence imaging, either during live-cell time-lapse experiments or after fixation. Image analysis can subsequently use nuclear signals to evaluate organization, division, migration, and lineage behavior.
This approach is useful when the experiment requires both cell tracking and a clear nuclear landmark. Because the signal is associated with chromatin, it can help identify individual nuclei while retaining spatial relationships within cell cultures or engineered tissues. It is therefore suited to studies of cellular organization, movement, division, and responses that unfold over time.