For the proof of principle cell motility separation assay, U373 Control cells were loaded with 5 μM CMFDA Cell Tracker dye (Thermo Fisher Scientific) and U373 DN Rac1 cells were pre-loaded with 5 μM CMPTX Cell Tracker dye (Thermo Fisher Scientific) per the manufacturer’s instructions. The two cell lines were mixed in a 1:1 ratio and a 30 μL aliquot of the mixed cell suspension (1 million cells mL−1) was seeded into the device. After 2 hours, the device was imaged with a FITC and TRITC filter cubes, in order to determine the starting locations of the cells within the channels. Then 30 μL of U373 media and U373 media containing 20 μg mL−1 SDF-1 were placed in opposing inlet ports. The aliquots were refreshed every 2 hours for a 10 hour period. After 10 hours, the device was again imaged to determine the final location of the cells.
Cmfda cell tracker dye
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom
CMFDA Cell Tracker dye is a fluorescent labeling reagent used for tracking and identifying cells. It binds to intracellular thiols, allowing for the visualization and monitoring of cell populations.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using cmfda cell tracker dye
1
Cell Motility Separation Assay
2
Visualizing Monocyte Adhesion to TNF-α-Activated Endothelial Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
hCMEC/D3 cells were cultured to confluency on transwell inserts (0.4-μm pore size, 0.33-cm2 diameter, Greiner Bio-One Gmbh, Austria) prior to 16 h of treatment with 10 ng/ml TNFα. Monolayers were then incubated for 2 h with U937 monocytic cells pre-labelled according to the manufacturer’s instructions with CMFDA cell tracker dye (ThermoFisher Scientific, UK). Co-cultures were washed vigorously with ice-cold PBS three times and fixed by incubation for 10 min in 1% formaldehyde in 0.1 M PBS. Co-cultures were mounted and examined using an Axiovert 200M inverted microscope (Zeiss) equipped with a 20x objective lens. Images were captured with ZEN imaging software (Carl Zeiss Ltd, UK) and analysed using ImageJ 1.53c (National Institutes of Health, USA).
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!