import tkinter as tk # Python 3
root = tk.Tk()

# The image must be stored to Tk or it will be garbage collected.
root.image = tk.PhotoImage(file='startup.gif')
label = tk.Label(root, image=root.image, bg='white')
root.overrideredirect(True)
root.geometry("+250+250")
root.lift()
root.wm_attributes("-topmost", True)
root.wm_attributes("-disabled", True)
root.wm_attributes("-transparentcolor", "white")
label.pack()
label.mainloop()