To provide a microscopic observation device, a microscopic observation method, or a manufacturing method of such a microscopic observation device that can perform an observation, using fluorescence from an observation target irradiated with excitation light.SOLUTION: A microscopic observation device 100, which irradiates an observation target with excitation light, and observes fluorescence generating from the observation target, comprises: a light source 1 that irradiates the observation target with the excitation light; a plurality of light reception units 2; a non-image formation lens system that guides the fluorescence from the observation target to each of the plurality of light reception units 2; and first polarization means 3 and second polarization means 4 that are arranged between the light source 1 and the light reception unit 2. Polarization characteristics of the first polarization means 3 and the second polarization means 4 are configured such that the excitation light passing through the first polarization means 3 is cut off by the second polarization means 4.SELECTED DRAWING: Figure 1
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