1. 正确转义路径字符串中的反斜杠.(或使用 r'raw string literal').

防止 PhotoImage 对象被垃圾收集.

Prevent PhotoImage object being garbage collected.

使用 file=... 选项指定文件名.

specify the filename using file=... option.

def start(root):
    startframe = tkinter.Frame(root)
    canvas = tkinter.Canvas(startframe,width=1280,height=720)

    startframe.pack()
    canvas.pack()

    # Escape / raw string literal
    one = tkinter.PhotoImage(file=r'images\one.gif')
    root.one = one  # to prevent the image garbage collected.
    canvas.create_image((0,0), image=one, anchor='nw')

更新

one = ... 和 root.one = one 两条语句可以合并为一条语句:

The two statements one = ... and root.one = one can be merged into one statement:

    root.one = one = tkinter.PhotoImage(r'images\one.gif')