What is CFSE labeling?
What is CFSE labeling?
CFSE is a versatile tool for the fluorescent intracellular labeling of live cells. Labeled cells can be assayed using flow cytometry and fluorescent microscopy. The dye is long lasting and well retained within labeled cells. The result is live cells with an intracellular fluorescent label.
How does CFSE staining work?
Unlike stains that label the lipid membrane of cells, CellTrace™ CFSE dye easily crosses the plasma membrane and covalently binds inside cells where the stable, well-retained fluorescent dye is designed to provide a consistent signal, even after several days in a cell culture environment.
What CFSE shows?
Carboxyfluorescein succinimidyl ester (CFSE) is an effective and popular means to monitor lymphocyte division1-3. The capacity of CFSE to label lymphocyte populations with a high fluorescent intensity of exceptionally low variance, coupled with its low cell toxicity, make it an ideal dye to measure cell division.
Why is CFSE used?
Carboxyfluorescein succinimidyl ester (CFSE) is a fluorescent cell staining dye. Subsequent studies revealed that the dye can be used to monitor lymphocyte proliferation, both in vitro and in vivo, due to the progressive halving of CFSE fluorescence within daughter cells following each cell division.
Is CFSE toxic to cells?
CFSE can be toxic to cells and non-optimal CFSE labeling conditions can thus hamper proliferation of cells and obscure interpretation of results. While CFSE is commonly used to assess lymphocyte proliferation, CFSE can be toxic and impair cell division.
What does CFSE stand for?
Carboxyfluorescein succinimidyl ester
Carboxyfluorescein succinimidyl ester (CFSE) is a fluorescent cell staining dye. CFSE is cell permeable and covalently couples, via its succinimidyl group, to intracellular molecules, notably, to intracellular lysine residues and other amine sources.
What is meant by Cfse?
The crystal field stabilization energy (CFSE) is the stability that results from placing a transition metal ion in the crystal field generated by a set of ligands. It arises due to the fact that when the d orbitals are split in a ligand field, some of them become lower in energy than before.
Can CFSE readily enter cells?
In our hands, we’ve found that lipophilic dyes such as PKH26 and PKH67 are particularly prone to transferring to adjacent cells in culture conditions; this is because they integrate into the cell membrane and cell membrane exchange occurs quite readily between cells.
How is CFSE used in the labeling of cells?
CFSE is a versatile tool for the fluorescent intracellular labeling of live cells. Labeled cells can be assayed using flow cytometry and fluorescent microscopy. The dye is long lasting and well retained within labeled cells. The provided CFSE is sufficient for ~1000 assays. CFSE is a cell permeant, non-fluorescent pro-dye.
How is CFSE divided in a flow cytometric analysis?
Covalently bound CFSE is divided equally between daughter cells, allowing discrimination of successive rounds of cell division. The techn … Flow cytometric analysis of cell division by dilution of CFSE and related dyes Curr Protoc Cytom. 2013;Chapter 9:Unit9.11.doi: 10.1002/0471142956.cy0911s64. Authors
Can a CFSE labeled cell be detected with FITC?
CFSE labeled cells can be detected with any instrument or filter set compatible with FITC detection: Excitation (max)=492nm, Emission (max)=517nm. Store at -20°C. Please refer to protocols.
How are HeLa cells labeled with CFSE ab113853?
One plate of HeLa cells was labeled with ab113853 (CFSE) and a second plate was treated with the hypoxia mimetic DFO but not labeled with CFSE. After harvesting, the cells were combined in a single tube, stained with a HIF1A antibody and subjected to flow cytometry. Expected results are shown in the left panel cartoon.