1. Field of the Invention
The present invention relates to a microscope apparatus.
2. Description of Related Art
Regarding a super-resolution microscope in the related art, STORM (Stochastic Optical Reconstruction Microscopy) is known (for example, refer to United States Patent Application, Publication No. 2008/0032414 and United States Patent Application, Publication No. 2008/0182336). In this microscope, a fluorescent material or a sample having a fluorescent material adhering thereto is used as an observed sample. This fluorescent material has a characteristic of being activated when irradiated with activation light having a predetermined wavelength and fluorescing to be inactivated when irradiated with excitation light having a different wavelength from the activation light later. A fluorescent material is activated with low density by irradiating an observed sample with weak activation light. By emitting excitation light later in order to make only the activated fluorescent material emit light, a fluorescent image is acquired. In the fluorescent image acquired in this manner, images of the fluorescent material that emits light with low density are separated individually. Accordingly, it is possible to obtain the position of the center of gravity of each image. By repeating such a step of obtaining the position of the fluorescent material multiple times, for example, hundreds of times to tens of thousands of times or more to create one image in which all images of the fluorescent material are disposed at their positions, it is possible to acquire a high-resolution observation image.
In addition, as a super-resolution microscope apparatus, a STORM based on a three-dimensional method capable of improving not only the resolution within the sample surface (XY plane) but also the resolution in the thickness direction (Z direction) of the sample is known (for example, refer to Bo Huang, et. al. Science 319, 810-813 (2008)). In such a STORM based on the three-dimensional method, the image of a fluorescent material can be made to have an elliptical shape by giving a predetermined astigmatic difference to the image of the sample by inserting a cylindrical lens in the imaging optical system, and the coordinate in the Z direction can be obtained from the ellipticity.