Super-resolution microscope system and method for illumination

A microscope system comprising:

• an adjusted specimen;
• and
• a microscope body;
• wherein said adjusted specimen is dyed with a molecule which has three electron states including at least a ground state;
• wherein said microscope body includes:
• a light source operable to provide light having a wavelength λ1 for exciting the molecule from the ground state to the first electron excited state;
• a light source operable to provide light having a wavelength λ2 for exciting the molecule in the first electron excited state to the second or higher electron excited state;
• a condensing optical system operable to condense the light having the wavelength λ1 and the light having the wavelength λ2 on said adjusted specimen;
• an overlap device operable to partially overlap an irradiation region of the light having the wavelength λ1 and an irradiation region of the light having the wavelength λ2 on said adjusted specimen;
• and
• an emission detector operable to detect an emission upon deexcitation of the excited molecule to the ground state;
• wherein a region of the emission upon deexcitation of the molecule from the first electron excited state to the ground state is inhibited by irradiating the light having the wavelength λ1 and the light having the wavelength λ2 through said overlap device;
• and
• wherein a beam obtained by condensing the light having the wavelength λ2 has a phase distribution in which the phase is shifted by π at a symmetric position with respect to an optical axis of the beam in a plane normal to the optical axis.

A method for illuminating an adjusted specimen using a microscope body, said method comprising:

• dying the adjusted specimen with a molecule which has three electron states including at least a ground state and which has an excited wavelength band from a first electron excited state to a second electron excited state which overlaps a fluorescent wavelength band upon deexcitation through a fluorescence process from the first electron excited state to a vibrational level in the ground state;
• providing light having a wavelength λ1 for exciting the molecule from the ground state to the first electron excited state;
• providing light having a wavelength λ2 for exciting the molecule in the first electron excited state to the second or higher electron excited state;
• condensing the light having the wavelength λ1 and the light having the wavelength λ2 on the adjusted specimen;
• partially overlapping an irradiation region of the light having the wavelength λ1 and an irradiation region of the light having the wavelength λ2 on the adjusted specimen;
• and
• detecting an emission upon deexcitation of the excited molecule to the ground state;
• inhibiting a region of the emission, upon deexcitation of the molecule from the first electron excited state to the ground state, by irradiating the light having the wavelength λ1 and the light having the wavelength λ2;
• and
• wherein a beam obtained by condensing the light having the wavelength λ2 has a phase distribution in which the phase is shifted by π a symmetric position with respect to an optical axis of the beam in a plane normal to the optical axis.

A method for illuminating an adjusted specimen using a microscope body, said method comprising:

• dying the adjusted specimen with a molecule which has three electron states including at least a ground state;
• providing light having a wavelength λ1 for exciting the molecule from the ground state to the first electron excited state;
• providing light having a wavelength λ2 for exciting the molecule in the first electron excited state to the second or higher electron excited state;
• condensing the light having the wavelength λ1 and the light having the wavelength λ2 on the adjusted specimen;
• partially overlapping an irradiation region of the light having the wavelength λ1 and an irradiation region of the light having the wavelength λ2 on the adjusted specimen;
• and
• detecting an emission upon deexcitation of the excited molecule to the ground state;
• inhibiting a region of the emission, upon deexcitation of the molecule from the first electron excited state to the ground state, by irradiating the light having the wavelength λ1 and the light having the wavelength λ2;
• and
• wherein a beam obtained by condensing the light having the wavelength λ2 has a phase distribution in which the phase is shifted by π at a symmetric position with respect to an optical axis of the beam in a plane normal to the optical axis.