Cfse Assay
Cfse Assay - Related Videos
09:49
Antigen Specific In Vivo Killing Assay using CFSE Labeled Target Cells
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Cited by 39 •
2010
Many infections elicit a strong CTL response, but occasionally, the quantity of responding cells does not correlate to control of the pathogen1. One measure of CTL quality is their ability to kill specifically2. CFSE labeling of target cells can be used to investigate this CTL response quality in vivo3,4.
04:10
The Use of Carboxyfluorescein Diacetate Succinimidyl Ester (CFSE) to Monitor Lymphocyte Proliferation
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Cited by 214 •
2010
CFSE covalently labels long-lived intracellular molecules with the fluorescent dye, carboxyfluorescein. As such, when a CFSE-labeled cell divides, its progeny have half the amount of fluorescence, which can thereby be used to assess cell division. This article describes the procedures typically used for labeling mouse lymphocytes with CFSE.
04:34
A CFSE-Based Assay to Monitor Skin Dendritic Cell Migration to Draining Lymph Nodes in Infected Mice
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2025
This video demonstrates an assay for monitoring dendritic cell migration following footpad infection with Mycobacterium bovis Bacille Calmette-Guérin BCG in mice. Upon bacterial infection, activated skin DC cells are labeled with a cell-permeable carboxyfluorescein diacetate succinimidyl ester (CFSE) dye. After a suitable interval for DC migration to the popliteal draining lymph node, the node cells are isolated and subjected to immunostaining for DC-specific surface markers. Identification of...
07:14
A CFSE-based Assay to Study the Migration of Murine Skin Dendritic Cells into Draining Lymph Nodes During Infection with Mycobacterium bovis Bacille Calmette-Guérin
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Cited by 10 •
2016
An assay in mice to track cell migration from skin to draining lymph node is described which enables the characterization of skin Dendritic cells mobilized to the lymph node after footpad infection with Bacille Calmette-Guérin.
11:54
Identification and Isolation of Slow-Dividing Cells in Human Glioblastoma Using Carboxy Fluorescein Succinimidyl Ester (CFSE)
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Cited by 18 •
2012
This video protocol demonstrates the application of the fluorescent dye carboxyfluorescein succinimidyl ester (CFSE) for the identification and separation of different sub-populations of cells in human glioblastoma based on frequency of cell division.
FAQs
CFSE first enters living cells, where intracellular esterases activate the dye. Its succinimidyl group then forms stable covalent bonds with cellular proteins, helping retain the fluorescent label within the cells. This chemical retention allows fluorescence to be followed through subsequent cell divisions and supports longitudinal measurement of proliferation in immune-cell populations.
When a labeled cell divides, its CFSE-associated fluorescence is distributed between the two daughter cells. Each successive division therefore leaves daughter cells with progressively lower fluorescence than their parent population. Flow cytometry detects these differences as sequential peaks, providing a pattern that can be used to quantify how many cells entered and progressed through division.
Progressive fluorescence loss indicates that labeled cells have undergone successive rounds of division rather than simply remaining in their original state. The degree and pattern of dilution therefore provide evidence of cellular proliferation. In immunology experiments, this readout helps determine whether immune cells responded after exposure to an antigen, vaccine, infection-related stimulus, or therapeutic compound.
Flow cytometry measures the fluorescence associated with individual cells and resolves the sequential peaks produced by dye dilution. This makes it possible to distinguish cells that retained stronger labeling from populations that divided repeatedly. The resulting profiles support quantitative assessment of proliferation and help identify which immune-cell populations responded under the tested experimental condition.
A typical workflow begins with living immune cells that take up CFSE. Intracellular esterases activate the dye, and the succinimidyl group binds cellular proteins. The labeled cells are then exposed to the experimental condition, such as an antigen, vaccine, infection-related setting, or therapeutic compound, before flow cytometry measures fluorescence patterns and division-associated dilution.
The assay is useful when researchers need to examine immune-cell proliferation after a defined biological exposure. In infection research, it can assess responses associated with infection or infection-related antigens. In broader immunology studies, the same approach supports evaluation of antigen responses, vaccine effects, and the influence of therapeutic compounds on immune activation.
CFSE profiles provide information about division history, not only the presence or number of cells. Sequential fluorescence peaks reveal progressively diluted labeling, allowing researchers to quantify proliferation across responding populations. Combined with flow-cytometric identification of those populations, the assay can show whether an experimental exposure produced measurable immune activation and which cells contributed to the response.