DCs were isolated as described above and rested overnight in RPMI media. The cells were then loaded with membrane-permeable Ca2+-sensitive fluorescent indicator Fluo-4 AM. Cells were incubated in Tyrode’s solution (in mM: NaCl 140, KCl 5.4, MgCl2 1, glucose 10, HEPES 10), containing 1 mM Ca2+, 6.6 µM fluo-4 AM, and 0.16% Pluronic F127 for 20 min at room temperature to load indicator in the cytosol. Then the supernatant was removed, and cells were washed in Tyrode’s solution, containing 1 mM Ca2+, twice for 15 min. After that, the cells were placed into the experimental chamber superfused with 2 mM Ca2+ Tyrode’s solution. Intracellular Ca2+ was measured using fluorescence photometry setup (IonOptix). Ca2+ fluorescence was first measured after 3–5 min in normal salt (140 mmol/L Na+), and then solution was changed to high salt (190 mmol/L Na) and Ca2+ fluorescence was monitored in 3-min intervals for another 21–24 min. All experiments were conducted at room temperature (≈23°C). Intracellular Ca2+ was analyzed using commercial software (IonWizard; IonOptix). A total of 7–12 DCs from nine different animals were used.
Fluorescence photometry setup
Manufactured by IonOptix
The Fluorescence Photometry Setup is a laboratory instrument designed to measure the fluorescence properties of samples. It provides accurate and reliable data on the intensity and spectral characteristics of fluorescent emissions from various materials and compounds.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using fluorescence photometry setup
1
Intracellular Calcium Dynamics in Dendritic Cells
2
Intracellular Calcium Dynamics in Dendritic Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
DCs were isolated as described above and rested overnight in RPMI media. The cells were then loaded with membrane-permeable Ca2+-sensitive fluorescent indicator Fluo-4 AM. Cells were incubated in Tyrode’s solution (in mM: NaCl 140, KCl 5.4, MgCl2 1, glucose 10, HEPES 10), containing 1 mM Ca2+, 6.6 µM fluo-4 AM, and 0.16% Pluronic F127 for 20 min at room temperature to load indicator in the cytosol. Then the supernatant was removed, and cells were washed in Tyrode’s solution, containing 1 mM Ca2+, twice for 15 min. After that, the cells were placed into the experimental chamber superfused with 2 mM Ca2+ Tyrode’s solution. Intracellular Ca2+ was measured using fluorescence photometry setup (IonOptix). Ca2+ fluorescence was first measured after 3–5 min in normal salt (140 mmol/L Na+), and then solution was changed to high salt (190 mmol/L Na) and Ca2+ fluorescence was monitored in 3-min intervals for another 21–24 min. All experiments were conducted at room temperature (≈23°C). Intracellular Ca2+ was analyzed using commercial software (IonWizard; IonOptix). A total of 7–12 DCs from nine different animals were used.
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!