Hi all!
As a part of my cryopreservation experiments, I m testing different vital stainings. I tried to check viability with FDA staining in 3 especies of filamentous brown algae (from active growing cultures) but it seems the cells don't catch the staining and, as a consequence, they do not produce the green fluorescence I expected or it is very low that I cannot notice it well. In addition, there is a green background fluorescence that complicates things more. I m using 0.01% of FDA (from an acetone solution stock of 1mg/mL stored at -20°C) dissolved in seawater. Incubation time on the dye is 20 minutes. I use the solution immediately.
Does anybody have any recommendation? I know FDA is used mainly on microalgae and protoplasts so I m not sure why it is not working in my seaweeds.
Thanks in advance!
Hello José Avila-Peltroche
You can use DAPI or CFSE stains to discern filamentous brown algae. The following link could be of help: Article Axenic clonal cultures of filamentous brown algae: Initiatio...
Regards,
Dr. Abhishek Mukherjee
I have used FDA successfully on filamentous algae (like Cladophora) in freshwater systems. I have noticed that sometimes FDA performs poorly in marine systems. It seems to be related to local water quality (maybe high pH?) which appears to make it less effective and often causes significant background fluorescence. We have shifted to using CMFDA in marine systems, sometimes in combination with FDA, and it seems to perform a bit better.
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