您可以使用互斥

bool firstInstance = true;
using (Mutex mutex = new Mutex(true, "MyApplicationName", out firstInstance))
{
    if (firstInstance)
    {
        Application.EnableVisualStyles();
        Application.SetCompatibleTextRenderingDefault(false);
        Application.Run(new MainForm());
    }
    else
    {
        // Another instance loaded
    }
}