Understanding copper sulphate decomposes at high temperature, can I supplement it to the media before autoclaving or should it be after autoclaving the media?
Copper sulfate (CuSO₄) is an essential micronutrient in plant tissue culture but is sensitive to high temperatures, as it can decompose and potentially lose its effectiveness when exposed to the heat of autoclaving. To ensure that it retains its bioavailability and doesn't degrade, **it's recommended to add copper sulfate after autoclaving the media**.
1. Autoclave your plant tissue culture media without the copper sulfate.
2. Prepare a sterile stock solution of copper sulfate at the desired concentration, sterilizing it by filter sterilization (using a 0.22 µm filter) rather than autoclaving.
3. After the media cools down (but before it solidifies if using agar), aseptically add the copper sulfate stock solution to the media under sterile conditions, such as in a laminar flow hood.
This approach helps maintain the integrity of the copper sulfate and ensures the proper availability of copper ions in the tissue culture medium.
The answer making by Raghad Mouhamad, our great colleage is perfect, because he explain step by step with very good reasons.
I don’t think the presence of copper play any strong role except it is act as cation in some enzyme synthesis. It presence at the rate 0.025 mg per litre is not much in the complex chemical reaction in the plant tissue culture. Definite it play crucial role as defence agent when use at higher conc.
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