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While this method addresses the issue of copper deficiency, further research is required to determine if it can also address excessive copper ion levels

For Safranin O/Fast Green staining, slides were stained with hematoxylin (Mayer, Sigma) for 5 min, 0.005% Fast Green (Thermo Fisher Scientific, Pittsburgh, PA, United States) for 1 min, rinsed with 1% acetic acid, and stained with 0.1% Safranin O (Millipore, Billerica, MA, United States) for 20 min

The LGA parameters were set as follows: population size (ga_pop_size) = A, energy evaluations (ga_num_generation) = B, mutation rate = C, crossover rate = D, step size = E, and number of LGA runs = 10