The invention discloses a novel pH response fluorescent molecular probe, which has a general formula of the structure as shown in the specification, wherein, n is equal to 1 to 5; R1 is an alkyl chain with 1 to 8 carbon atoms; and R2 is hydrogen, fluorine, chlorine, bromine, iodine, methoxy, ethoxy or nitro connected to the 4th, 5th, 6th, 7th, 4'th, 5'th, 6'th or 7'th site of a benzimidazole ring. The invention also discloses an application of the novel fluorescent molecular probe in the field of pH response fluorescent detection. The novel pH response fluorescent molecular probe has very wide detection wavelength, and can be excited from ultraviolet to visible light (less than 520nm). Fluorescent images of the novel fluorescent molecular probe in different areas of a life system can also be acquired in combination with confocal laser scanning microscopy. Compared with the existing pH response fluorescent molecular probe, the novel pH response fluorescent molecular probe has the advantages that the preparation method is simple and the displacement value with exciting light is large.