Hi,
In literature I noticed that who stains haemocytes with FDA for 30 min then stop the reaction in ice while this does not apply for 15 min incubation time (or maybe authors did not write this particular).
There is any real necessity? Can the dye adverse interact with the cells?
Thanks in advance.
Best,
Elisabetta
Hello Maria
FDA is a non-fluorescent lipid soluble dye which is a substrate for esterase enzyme measuring enzyme activity by getting hydrolysed to fluorescein after cellular uptake. This fluorescein retained by intact cells gives the fluorescence. So FDA measures both esterase activity as well as membrane integrity of the cells.
The entire reaction has to be performed in ice because it involves the measurement of esterase enzyme.
Enzymes are temperature and pH sensitive.
Regards.
Hello, Maria!
You look also at this chapter:
Article Quantitative chracterization of viability and growth dynamic...
The signal given is nonsense. Higher above a certain level,it is not worth measuring. You measure the quantity of light a certain time. You may stop reactions with H2SO4 and it is the same principle. You measure the kind of light and then you stop measuring
- Badges
- Science method
- Similar topics
- Biological Science
- Histology
- Staining