For my work, I am investigating the effect of organophosphate pesticide on stress and anxiety-like behaviours and the associated neurotransmitter levels (which includes serotonin). To estimate the serotonin in the zebrafish brain, I dissected and took two zebrafish brains and homogenized the tissue with PBS containing 10^-4 M ascorbic acid. And I estimated the serotonin levels with a commercially available kit (Serotonin ELISA kit, Bioassay Technologies, Cat. no. E1128Hu). Based on the calibration curve I found my sample to be containing serotonin levels (for two brains 175.6 ng and for three brain samples to be 220.66 ng, R square value = 0.9945). I would like to know whether these levels fall within the physiological range? or do I need to normalize these values with the sample weight or sample protein content. Please help me in this regard, I have also attached my data sheet for your reference.
Hi Christy Lite,
In my view, you need to correct (normalize) those values to the sample weight. This will give you normalized values that could be compared to other works. This is especially important since brain removal could affect the amount of tissue that you have at the beginning of your assay. Unfortunately I do not have data from zebrafish to compare with your values.
