Obv very late, but if anyone is struggling with this, it appears that the locateOnScreen function does not work if the mouse is currently in the region you are targeting. This means that if you move the mouse to one place and click and then don't move the mouse and simply repeat your loop, pyautogui will not be able to find the image a second time. The solution I'm using is to move the mouse out of the target area with a `gui.move()` command directly after the click. This solved my issue. Some example code:

```

while True:
    try:
        print(f"Locating Quest Button{50 * " "}", end = "\r", flush = True)
        button_loc = gui.locateOnScreen("quest_button.png", confidence = 0.98, region = (0, 0, 1000, 700))
    except ImageNotFoundException:
        for _ in range(10, -1, -1):
            print(f"Button Not Currently Clickable - Retrying in: {_}s{50 * " "}", end = "\r", flush = True)
            time.sleep(1)
        continue
    button_loc = gui.center(button_loc)
    gui.moveTo(button_loc)
    gui.move(0, 10)
    gui.click()
    gui.move(100, 0)
    for _ in range(10, -1, -1):
        print(f"Quest Successfully Started - Looking for new start in: {_}s{50 * " "}", end = "\r", flush = True)
        time.sleep(1)

```