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Two stronger areas in the changes of SERS intensity were achieved with a high concentration of halogen ions, while only one maximum intensity area was obtained with a low concentration of halogen ions added. This method can effectively avoid complex and unpredictable microenvironments of SERS substrates in the liquid drop, further improving the reproducibility of SERS detection as well as broadening it to biological applications.Hill coefficients, which provide a measure of cooperativity in ligand binding, can be determined for equilib