Can you tell us that what is purpose of you reseach ? thank you very much 

Hi Vesela,

I assume you will use it as a stain in isolated living cells to check for vitality and lysosomal alterations e.g. blood to assess neutrophils for the so - called toxic lysosomes. Am I right?

Best regards,

Ivan

P.S. Можем да продължим на български също :)

I work with fish from metal-contaminated aquatic ecosystems and mainly perform histological analysis. However, I am also interested in lysosmal alterations. I have once worked with blue mussels and I know the method which is applied for them but I have no idea how to adapt it for fish. Thanks a lot for your concern. Cheers.

I am not familiar with the neutral red retention assay, but I have been briefly involved with lysosomal stability assays. One assay that can be done without any genetic manipulation of your system is the "acridine orange lysosome explosion assay" from Kirkegaard et al Nature 2010 (http://www.ncbi.nlm.nih.gov/pubmed/20111001). 

Another approach would be to study the sensitivity of your cells to drugs that are known to cause lysosomal cell death, the so called cationic amphiphillic drugs CADs, see Petersen et al Cancer Cell 2013 (http://www.sciencedirect.com/science/article/pii/S153561081300353X).

In the latter you would just add these drugs and see if there is more cell death in your cells exposed to toxic environments. This is an easier assay, but the interpretation of the results is more difficult because the read-out is more indirect.

Thank you, Petter. The information which you provided will be very helpful.