Dear All,
I am encountering challenges with AAV-mediated neuronal transduction, where high volumes lead to significant cell death, while low volumes yield inadequate signal levels.
Here's a brief overview of my AAV production process:
For titration, I've tried volumes ranging from 10 microliters to 0.2 microliters. At 1 microliter, the signal is optimal, but it coincides with significant cell death. Volumes lower than this are not feasible for analysis due to inadequate signal.
I'm seeking insights into the possible reasons behind the observed cell death during transduction. Your input would be greatly appreciated.
it contains a glutamate sensor fused with a synapse localisation signal. Another one contains a calcium sensor without anyfusion.
Both transgenes are known not to be toxic for neurons.
Hey,
I prepared the buffer.
When I transduce, my neurons are in neurobasal phenolred negative, I cannot check colour change.
But I tried only buffer titration from 1 microliter to 50 microliter. It made no cell death.
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