Microscope system and method for measuring refractive index of sample
View Patent ↗A microscope system includes: a wavefront modulator that modulates a wavefront of light from a light source; an objective that irradiates a sample with light whose wavefront has been modulated by the wavefront modulator; a spherical aberration corrector that corrects spherical aberration caused by a difference between a refractive index of a medium between the objective and the sample and a refractive index of the sample; a refractive index calculator that calculates, for each wavelength of the light from the light source, an average refractive index of a medium between the objective and a condensing position of light emitted from the objective on the basis of an amount of the corrected spherical aberration; and a controller that controls the wavefront modulator to correct chromatic aberration calculated on the basis of the calculated average refractive index for each wavelength.
1. A microscope system comprising:
a wavefront modulator that modulates a wavefront of light from a light source;
an objective that irradiates a sample with light whose wavefront has been modulated by the wavefront modulator;
a spherical aberration corrector that corrects spherical aberration caused by a difference between a refractive index of a medium between the objective and the sample and a refractive index of the sample;
a refractive index calculator that calculates, for each wavelength of the light from the light source, an average refractive index of a medium between the objective and a condensing position of light emitted from the objective on the basis of an amount of the spherical aberration corrected by the spherical aberration corrector; and
a controller that controls the wavefront modulator to correct chromatic aberration calculated on the basis of the average refractive index for each wavelength that has been calculated by the refractive index calculator.
2. The microscope system according to claim 1 , further comprising:
a chromatic aberration calculator that calculates chromatic aberration generated between wavelengths on the basis of the average refractive index for each wavelength that has been calculated by the refractive index calculator,
wherein the controller controls the wavefront modulator to correct the chromatic aberration calculated by the chromatic aberration calculator.
3. The microscope system according to claim 1 , further comprising:
a Z scanner that relatively moves the objective in an optical axis direction with respect to the sample, and changes the condensing position of the light emitted from the objective,
wherein the refractive index calculator calculates the average refractive index of the medium between the objective and the condensing position of the light emitted from the objective on the basis of the amount of the spherical aberration corrected by the spherical aberration corrector, for each of the condensing positions changed by the Z scanner and for each wavelength of the light from the light source.
4. The microscope system according to claim 1 , wherein the wavefront modulator modulates a wavefront of light having one of two wavelengths under the control of the controller such that a condensing position of the light having the one of the two wavelengths moves in an optical axis direction of the objective by a distance represented by the chromatic aberration generated between the two wavelengths that has been calculated by the refractive index calculator.
5. The microscope system according to claim 1 , wherein the objective includes a correction collar that is the spherical aberration corrector.
6. The microscope system according to claim 1 , further comprising:
a multi-photon excitation microscope that includes the objective, the wavefront modulator, and the spherical aberration corrector.
7. A method for measuring a refractive index of a sample with a microscope system, the method comprising:
relatively moving an objective in an optical axis direction with respect to the sample so as to make a condensing position of light emitted from the objective of the microscope system coincide with a surface of the sample;
relatively moving the objective in the optical axis direction with respect to the sample so as to move the condensing position of the light emitted from the objective inside the sample;
correcting spherical aberration inside the sample; and
calculating, by a refractive index calculator of the microscope system, the refractive index of the sample between the surface of the sample and the condensing position inside the sample on the basis of an amount of the corrected spherical aberration,
wherein the calculating the refractive index of the sample by the refractive index calculator comprises:
calculating an average refractive index of a medium between the objective and the condensing position inside the sample on the basis of the amount of the corrected spherical aberration; and
calculating a refractive index of the sample between the surface of the sample and the condensing position inside the sample on the basis of a working distance of the objective, a movement amount of the condensing position from the surface of the sample to the inside of the sample, the calculated average refractive index, and a refractive index of a medium between the objective and the surface of the sample.