An auto-focusing method for determining an in-focus position of a plurality of wells in at least a portion of a multi-well plate, the method including using a first objective lens having a first magnification to identify, in each of at least three wells of a selected subset of the plurality of wells, an in-focus position of each well with respect to the first objective lens, on the basis of at least three the in-focus positions, computing a plane along which the at least three wells will be in focus with respect to at least one objective lens having a second magnification that is not greater than the first magnification, and using the at least one objective lens to scan, along the plane, at least some of the plurality of wells in the portion of the plate.

The present invention relates to the high resolution imaging of samples using imaging mass spectrometry (IMS) and to the imaging of biological samples by imaging mass cytometry (IMCTM) in which labelling atoms are detected by IMS. LA-ICP-MS (a form of IMS in which the sample is ablated by a laser, the ablated material is then ionised in an inductively coupled plasma before the ions are detected by mass spectrometry) has been used for analysis of various substances, such as mineral analysis of geological samples, analysis of archaeological samples, and imaging of biological substances. However, traditional LA-ICP-MS systems and methods may not provide high resolution. Described herein are methods and systems for high resolution IMS and IMC.

The present invention relates to the high resolution imaging of samples using imaging mass spectrometry (IMS) and to the imaging of biological samples by imaging mass cytometry (IMCTM) in which labelling atoms are detected by IMS. LA-ICP-MS (a form of IMS in which the sample is ablated by a laser, the ablated material is then ionised in an inductively coupled plasma before the ions are detected by mass spectrometry) has been used for analysis of various substances, such as mineral analysis of geological samples, analysis of archaeological samples, and imaging of biological substances. However, traditional LA-ICP-MS systems and methods may not provide high resolution. Described herein are methods and systems for high resolution IMS and IMC.

A lens for a still or film camera includes a first and second lens-element arrangement, and a wavefront manipulator. The first and second lens-element arrangement are arranged spaced mutually apart along an optical axis of the lens such that an interstice is present therebetween. The wavefront manipulator is situated in the interstice and includes at least two optical components which are arranged so as to be displaceable counter to one another, perpendicular to the optical axis, and which each include a free-form surface. The wavefront manipulator has a zero position, in which the optical components thereof do not cause any image aberrations in the imaging properties of the lens, and effective positions, in which the optical components are displaced counter to one another, out of the zero position perpendicular to the optical axis, and in which the optical components cause a spherical aberration in the imaging properties of the lens.

A method for improving the lateral resolution of fluorescence microscopy using inhomogeneous surface wave microscopy is provided. The microscope includes a prism on which laterally-interfaced plasmonic nanofilms are deposited (here called metal 1 and metal 2, though materials other than metals may be used, see Claim 1). A propagating wave which has evanescent character along one spatial dimension, known as a surface plasmon polariton, is excited on the first metal nanofilm by focusing of monochromatic incident light with a particular incident angle through the prism. Propagation of the surface plasmon polariton across the interface between the metal 1 nanofilm and the metal 2 nanofilm creates a propagating wave with evanescent character in two spatial dimensions, known as an inhomogeneous surface plasmon polariton [3]. A key property of inhomogeneous surface plasmon polaritons is the external controllability of the evanescent character of the wave in both the axial and lateral dimensions, which imparts the ability to judiciously enhance lateral resolution of conventional total internal reflection fluorescence microscopy with only minor modifications to the device.

A method for improving the lateral resolution of fluorescence microscopy using inhomogeneous surface wave microscopy is provided. The microscope includes a prism on which laterally-interfaced plasmonic nanofilms are deposited (here called metal 1 and metal 2, though materials other than metals may be used, see Claim 1). A propagating wave which has evanescent character along one spatial dimension, known as a surface plasmon polariton, is excited on the first metal nanofilm by focusing of monochromatic incident light with a particular incident angle through the prism. Propagation of the surface plasmon polariton across the interface between the metal 1 nanofilm and the metal 2 nanofilm creates a propagating wave with evanescent character in two spatial dimensions, known as an inhomogeneous surface plasmon polariton [3]. A key property of inhomogeneous surface plasmon polaritons is the external controllability of the evanescent character of the wave in both the axial and lateral dimensions, which imparts the ability to judiciously enhance lateral resolution of conventional total internal reflection fluorescence microscopy with only minor modifications to the device.

The solid immersion lens unit includes: a solid immersion lens having a contact surface allowed to be in contact with an inspection object and a spherical surface allowed to be opposite to an objective lens; a holder holding the solid immersion lens; a magnet provided to the holder; and a spherical body rotatably held by a magnetic force of the magnet at a position opposite to the spherical surface. The holder swingably holds the solid immersion lens in a state where the spherical surface is in contact with the spherical body.

The solid immersion lens unit includes: a solid immersion lens having a contact surface allowed to be in contact with an inspection object and a spherical surface allowed to be opposite to an objective lens; a holder holding the solid immersion lens; a magnet provided to the holder; and a spherical body rotatably held by a magnetic force of the magnet at a position opposite to the spherical surface. The holder swingably holds the solid immersion lens in a state where the spherical surface is in contact with the spherical body.

A light sheet microscope includes a sample chamber in which a cover slip or slide is arrangeable, which has a surface that defines a partially reflective interface and which has a further surface that defines a further partially reflective interface. The two interfaces are arranged at different distances from an objective. The light sheet microscope further includes an optical system having the objective facing toward the cover slip or slide, an illumination apparatus, which is designed to generate a light sheet, a sensor, and a processor. The two interfaces are formed in that two optical media are applicable in the sample chamber. The light sheet microscope forms a measuring device for acquiring a measured variable. The sensor is designed to acquire the intensities and/or the incidence locations of the two reflection light beams.

A light sheet microscope includes a sample chamber in which a cover slip or slide is arrangeable, which has a surface that defines a partially reflective interface and which has a further surface that defines a further partially reflective interface. The two interfaces are arranged at different distances from an objective. The light sheet microscope further includes an optical system having the objective facing toward the cover slip or slide, an illumination apparatus, which is designed to generate a light sheet, a sensor, and a processor. The two interfaces are formed in that two optical media are applicable in the sample chamber. The light sheet microscope forms a measuring device for acquiring a measured variable. The sensor is designed to acquire the intensities and/or the incidence locations of the two reflection light beams.