The present invention relates, in general, to a
fluorescence microscope and method of observing samples using the
microscope and, more particularly, to a
fluorescence microscope and method of observing samples using the microscope, which can reduce
optical noise and obtain images with higher sensitivity, thus obtaining precise information about the density, quantity, location, etc. of a
fluorophore, and which can simultaneously process separate images even when a plurality of fluorophores having different excitation and fluorescent
wavelength ranges is distributed, thus easily obtaining information about the fluorophores. The
fluorescence microscope of the present invention includes an objective lens, and first and third medium units. The first medium unit has a
refractive index of n1 to accommodate one or more micro-objects including fluorophores and provide a path of excitation light to excite the fluorophores. The third medium unit has a
refractive index of n3, and is placed between the first medium unit and the objective lens to totally reflect the excitation light incident through the first medium unit at an interface of the third medium unit coming into contact with the first medium unit. The refractive indices of the third and first medium units satisfy a relationship of n1>n3.