Carboxyfluorescein succinimidyl ester (cfse)
Manufactured by Olympus
Sourced in Japan
CFSE (Carboxyfluorescein Succinimidyl Ester) is a fluorescent dye used for cell labeling and tracking in various biological applications. It binds to intracellular proteins, allowing the visualization and monitoring of cell division and proliferation in cell culture experiments.
Automatically generated - may contain errors
Lab products found in correlation
Pkh26, by Olympus (1 mentions)
Pkh26, by Merck Group (1 mentions)
Carboxyfluorescein succinimidyl ester (cfse), by Merck Group (1 mentions)
Fv1200, by Olympus (1 mentions)
Hexidium iodide, by Thermo Fisher Scientific (1 mentions)
Fv3000, by Olympus (1 mentions)
Carboxyfluorescein succinimidyl ester (cfse), by Thermo Fisher Scientific (1 mentions)
Syto 11, by Thermo Fisher Scientific (1 mentions)
Bx 15 microscope, by Olympus (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Sybr green supermix, by Bio-Rad (1 mentions)
Facscalibur flow cytometer, by BD (1 mentions)
Carboxyfluorescein succinimidyl ester (cfse), by BD (1 mentions)
Fluorescence microscope, by Olympus (1 mentions)
5 protocols using carboxyfluorescein succinimidyl ester (cfse)
1
Fluorescent Labeling of sEVs
2
Visualizing Bacterial Coaggregation
Check if the same lab product or an alternative is used in the 5 most similar protocols
F. nucleatum subsp. polymorphum and S. gordonii were cultured to the late-exponential phase. Bacterial cells were washed three times and resuspended in sterile PBS. For visualization, F. nucleatum subsp. polymorphum was stained green with 5-(and-6)-carboxyfluorescein succinimidyl ester (CFSE) (Thermo Fisher, USA), while S. gordonii was stained red with hexidium iodide (Thermo Fisher, USA) according to the manufacturer’s instructions. Samples were incubated for 15 min in darkness at room temperature. Fluorescently stained bacteria were washed three times with sterile PBS and resuspended in CAB. The coaggregated F. nucleatum subsp. polymorphum and S. gordonii (Fnp-Sg) were obtained as described above. Coculture of the two species (Fnp+Sg) in PBS, where they did not coaggregate with each other but only mixed physically, were used as controls. After coaggregation reactions, 10 μL of coaggregated Fnp-Sg was transferred to a glass slide and covered with a cover glass. The coaggregation and coculture samples were visualized by an Olympus confocal microscope (FV3000, Olympus, Japan) using excitation (Ex) at 492 nm and emission (Em) at 517 nm for CFSE and Ex/Em = 518 nm/600 nm for hexidium iodide.
+ Expand
F. nucleatum subsp. polymorphum and S. gordonii were cultured to the late-exponential phase. Bacterial cells were washed three times and resuspended in sterile PBS. For visualization, F. nucleatum subsp. polymorphum was stained green with 5-(and-6)-carboxyfluorescein succinimidyl ester (CFSE) (Thermo Fisher, USA), while S. gordonii was stained red with hexidium iodide (Thermo Fisher, USA) according to the manufacturer’s instructions. Samples were incubated for 15 min in darkness at room temperature. Fluorescently stained bacteria were washed three times with sterile PBS and resuspended in CAB. The coaggregated F. nucleatum subsp. polymorphum and S. gordonii (Fnp-Sg) were obtained as described above. Coculture of the two species (Fnp+Sg) in PBS, where they did not coaggregate with each other but only mixed physically, were used as controls. After coaggregation reactions, 10 μL of coaggregated Fnp-Sg was transferred to a glass slide and covered with a cover glass. The coaggregation and coculture samples were visualized by an Olympus confocal microscope (FV3000, Olympus, Japan) using excitation (Ex) at 492 nm and emission (Em) at 517 nm for CFSE and Ex/Em = 518 nm/600 nm for hexidium iodide.
3
Quantifying Trypanosoma cruzi Infection
Check if the same lab product or an alternative is used in the 5 most similar protocols
T. cruzi trypomastigotes were labeled with 5 µM SYTO®11 (binds DNA, Molecular Probes-Invitrogen, Eugene, OR) or 5 µM carboxyfluorescein succinimidyl ester (CFSE, binds amines, Invitrogen) for 20 min at 37oC. THP-1- or BM- derived mφs were infected and incubated with labeled T. cruzi trypomastigotes, as above. Cells were washed, and SYTO®11 or CFSE fluorescence as an indicator of parasite uptake was determined by using an Olympus BX-15 microscope equipped with a digital camera (magnification 40X). Cells infected with CFSE-labeled parasites were also fixed with 2% paraformaldehyde and visualized on a FACSCalibur flow cytometer (BD Biosciences, San Jose, CA) acquiring 20,000 events. Further analysis was performed by using FlowJo software (ver. 7.6.5, Tree-Star, San Carlo, CA). Mean Fluorescence intensity (MFI) of CSFE positive cells was used as a relative marker of parasites per cell.
Total DNA from normal and infected cells was isolated by using TRIzol reagent (Life Technologies, Grand Island, NY). Total DNA (100 ng) was used as a template in a quantitative PCR (qPCR) on an iCycler thermal cycler with SYBR Green Supermix (Bio-Rad) and oligonucleotide pairs specific for Tc18S ribosomal DNA (Table S1 ). Data were normalized to murine or human GAPDH, and fold change calculated as 2−ΔΔCt, where ΔΔCt represents the Ct (sample) - Ct (control).
+ Expand
T. cruzi trypomastigotes were labeled with 5 µM SYTO®11 (binds DNA, Molecular Probes-Invitrogen, Eugene, OR) or 5 µM carboxyfluorescein succinimidyl ester (CFSE, binds amines, Invitrogen) for 20 min at 37oC. THP-1- or BM- derived mφs were infected and incubated with labeled T. cruzi trypomastigotes, as above. Cells were washed, and SYTO®11 or CFSE fluorescence as an indicator of parasite uptake was determined by using an Olympus BX-15 microscope equipped with a digital camera (magnification 40X). Cells infected with CFSE-labeled parasites were also fixed with 2% paraformaldehyde and visualized on a FACSCalibur flow cytometer (BD Biosciences, San Jose, CA) acquiring 20,000 events. Further analysis was performed by using FlowJo software (ver. 7.6.5, Tree-Star, San Carlo, CA). Mean Fluorescence intensity (MFI) of CSFE positive cells was used as a relative marker of parasites per cell.
Total DNA from normal and infected cells was isolated by using TRIzol reagent (Life Technologies, Grand Island, NY). Total DNA (100 ng) was used as a template in a quantitative PCR (qPCR) on an iCycler thermal cycler with SYBR Green Supermix (Bio-Rad) and oligonucleotide pairs specific for Tc18S ribosomal DNA (
4
Cell Proliferation on Electrode Surface
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
In order to assess the proliferation of cells on the electrode surface, MPC were stained with 5(6)-carboxyfluorescein N-hydroxysuccinimidylester (CFSE; Abcam, Cambridge, UK) according to the protocol provided by the manufacturer. Briefly, cells were cultured at standard cell conditions (37 °C, 5 % CO2) until they reached confluence (cell number approximately 1 × 106). Cells were then trypsinised and re-suspended several times in fresh culture medium. Thereafter, cells were centrifuged at 800 rpm for 4 min and the supernatant was discarded. Cells were re-suspended with 1 ml PBS containing 10 μM CFSE and were allowed to rest in the solution for 10 min at room temperature. To stop the staining procedure, 1 ml of serum containing MNC medium was added to the cell suspension and pipetted several times prior to centrifugation for the removal of unbound CFSE. Finally, cells were re-suspended with culture medium and the success of the CFSE staining was evaluated using a fluorescence microscope (Olympus, Shinjuku, Japan).
Coating of the electrodes was performed as described above using the stained cells and cultivated for 10 days. Fluorescence microscopy was applied on the 3rd, 7th, and 10th day after coating. Therefore, the medium was replaced with pre-heated PBS and the dye was excited with a light and was detected using the standard FITC filter of the microscope.
Coating of the electrodes was performed as described above using the stained cells and cultivated for 10 days. Fluorescence microscopy was applied on the 3rd, 7th, and 10th day after coating. Therefore, the medium was replaced with pre-heated PBS and the dye was excited with a light and was detected using the standard FITC filter of the microscope.
5
Evaluating DPSC Viability on ZIF-8
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Carboxyfluorescein diacetate succinimidyl ester (CFSE; Sigma, Munich, Germany) was used to evaluate the viability of cultured cells on the ZIF-8 substrate. After one day, the attached cells were incubated with serum-reduced (1% FBS) medium contained five µM CFSE for 30 minutes. Next, the esterase reaction was quenched by adding serum-supplemented (10%) medium, and dead cells were stained by five µg/mL propidium iodide (PI) for one minute. Imaging by fluorescence microscopy (Olympus, BX51, Japan) showed live DPSCs as green CFSE labeled cells, while dead cells were counterstained with PI.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!