The invention discloses a single-cell / single-molecule imaging light / electricity comprehensive tester based on a multifunctional probe. The tester comprises the multifunctional nano-probe, a sample pool, an atomic force microscope system, a light source unit, an electricity testing unit, a cell sample locating system and a photon testing unit. The light source unit, the electricity testing unit, the cell sample locating system and the photon testing unit are respectively connected with a data collection and analyzing unit. The multifunctional nano-probe comprises an optical fiber layer, a nano electrode layer and an insulation layer. The nano electrode layer and the insulation layer are wrapped on the outer wall of the optical fiber layer in sequence. The synchronization comprehensive tester comprises a manufacturing method of the nano-probe which can carry out atomic force scanning imaging on single-cells / single-molecules and carry out light / electricity testing at the same time, a manufacturing method of the sample pool and relevant optics, electricity and mechanics testing and analyzing instruments. Temporal-spatial resolution and the range of target objects which can be tested are greatly improved, the tester can be widely used in biology and medicine and be used for fundamental research and disease testing under the level of sub-cells and molecules, and the tester has great development significance on tumor early diagnosis, medicine development and agricultural science.