Sample image acquisition device and sample image generation device

A sample image acquisition device comprising:

• a first optical system configured to form a first optical image of a sample;
• a second optical system configured to form a second optical image of the sample;
• a moving mechanism;
• and
• an image generation unit, wherein
• the first optical system includes an observation optical system positioned in an optical path between the sample and the first optical image,
• an optical axis of the first optical system and an optical axis of the second optical system coincide at a position of the sample,
• the moving mechanism changes a distance between the sample and a first objective lens of the first optical system and a distance between the sample and a second objective lens of the second optical system,
• a fluorescent image group including a plurality of fluorescent images and a bright field image group including a plurality of bright field images are generated by changing the distances,
• the fluorescent image group is generated based on the first optical image,
• the bright field image group is generated based on the second optical image,
• an estimation sample is a modeled sample of the sample,
• a virtual observation optical system is a modeled optical system of the observation optical system,
• the image generation unit includes
• a memory configured to store the fluorescent image group and the bright field image group, and
• a processor, and
• the processor
• calculates an estimation image of the estimation sample using a refractive index distribution of the estimation sample,
• calculates, as a final refractive index distribution, the refractive index distribution optimized using each of the bright field images and the estimation image,
• divides each of the fluorescent images into a plurality of areas,
• sets a virtual area corresponding to each of the areas in the estimation sample,
• sets a virtual light beam that travels from the virtual area toward the virtual observation optical system,
• calculates a point spread function corresponding to the area, using the final refractive index distribution included in a range in which the virtual light beam travels,
• generates, for each of the areas, an image corresponding to the area using the point spread function and the fluorescent image of the area,
• combines all of the images to generate an image corresponding to the fluorescent image as a final image,
• executes calculation of the final refractive index distribution for each image in the bright field image group, and
• executes generation of the final image for each image in the fluorescent image group.