The invention provides an optical super-resolution microscopic imaging system comprising a dichroic beamsplitter for annular parallel light to transmit through; a focusing lens used for converging the annular parallel light transmitted through the dichroic beamsplitter; a confocal pinhole for the annular parallel light after being converged to pass through to filter the annular parallel light; a varifocal lens system for collimating the annular parallel light passing through the confocal pinhole into excited annular parallel light; and a detector for receiving and processing fluorescence emitted by the excited sample, the fluorescence emitted by the excited sample being returned by the same way, and the dichroic beamsplitter separating the fluorescence emitted by the sample from an annular parallel light path and turning the fluorescence to the detector to obtain a super-resolution image of the sample.

An anisoplanatic aberration correction method and apparatus for adaptive optical linear beam scanning imaging. The method comprises: in an adaptive optical linear beam scanning imaging system, performing temporal correction on an anisoplanatic region aberration in a linear beam scanning direction, and performing regional correction on an anisoplanatic region aberration in a linear beam direction. According to the method, the limitation of an isoplanatic region on an adaptive optical imaging field of view can be overcome, and wide field of view aberration correction and high-resolution imaging of a retina is realized. According to the provided method and apparatus for temporal and regional correction of a wide field of view anisoplanatic aberration, the wide field of view aberration correction can be completed by means of only a single wavefront sensor and a single wavefront corrector, such that almost none of the system complexities is increased. The provided correction of an image subjected to deconvolution is low in cost. By means of regional deconvolution of wavefront aberration information, the adaptive optical aberration correction can be compensated to the greatest possible extent, the correction effect is good, and online processing or post-processing can be performed, and correction is flexible and convenient.

In high-resolution scanning microscopy, a sample is excited by illumination radiation to emit fluorescence radiation in such a way that the illumination radiation is focused at a point in or on the sample to form a diffraction-limited illumination spot. The point is imaged in a diffraction-limited manner into a diffraction image on a spatially resolving surface detector, wherein the surface detector has a spatial resolution that resolves a structure of the diffraction image. The sample is scanned by means of different scanning positions with an increment of less than half the diameter of the illumination spot. An image of the sample is generated from the data of the surface detector and from the scanning positions assigned to said data, said image having a resolution that is increased beyond a resolution limit for imaging. For the purposes of distinguishing between at least two predetermined wavelength regions in the fluorescence radiation from the sample, a corresponding number of diffraction structures are generated on the surface detector for the at least two predetermined wavelength ranges, said diffraction structures differing but having a common center of symmetry. The diffraction structures are evaluated when generating the image of the sample.

A method for training a mathematical model which describes a light propagation in a reflection microscopy includes radiating a light distribution I.sub.0 into an excitation path of a microscope, modulating the light distribution I.sub.0 to form a light distribution I.sub.A in the excitation path via an optical modulator, reflecting the light distribution I.sub.A at a location of a sample in a detection path of the microscope, modulating the light distribution I.sub.A to form a light distribution I.sub.D in the detection path via a further optical modulator, recording a reflected light distribution I.sub.D, repeating the above steps n-fold to generate an n-fold 3-tuple (M.sub.A, M.sub.D; I.sub.D), transferring the n-fold 3-tuple (M.sub.A, M.sub.D; I.sub.D) to a computer to implement a mathematical model F for a light propagation in reflection microscopy, and ascertaining the mathematical model F which describes the light propagation in reflection microscopy based on the n-fold 3-tuple (M.sub.A, M.sub.D; I.sub.D).

A super-resolution microscope avoids the need for complex phase plate optics normally used to produce a doughnut-shaped depletion beam by employing low-intensity regions of common diffraction patterns such as an Airy disk.

An optical method of measurement and an optical apparatus for determining the spatial position of at least one luminous object on a sample. A sequence of at least two compact luminous distributions of different topological families is projected onto the sample, and light re-emitted by the at least one luminous object is detected. At least one optical image is generated for each luminous distribution on the basis of the light detected. The optical images are analyzed to obtain spatiotemporal information regarding the light re-emitted by the at least one luminous object, or location of the at least one luminous object.

A method for operating a light microscope with structured illumination includes: providing illumination patterns by means of a structuring device which splits impinging light into at least three coherent beam parts which correspond to a −1., 0., and +1. diffraction orders of light; generating different phases of the illumination patterns by setting different phase values for the beam parts with phase shifters; and recording at least one microscope image for each of the illumination patterns and calculating a high resolution image from the microscope images. Phase shifters are provided not only for the beam parts of the −1. and +1. diffraction orders but also at least one phase shifter for the beam part of the 0. diffraction order. At least two different phase values Φ.sub.0 are set with the at least one phase shifter for the 0. diffraction order to provide a plurality of illumination patterns with different phases.

Techniques for acquiring focused images of a microscope slide are disclosed. During a calibration phase, a “base” focal plane is determined using non-synthetic and/or synthetic auto-focus techniques. Furthermore, offset planes are determined for color channels (or filter bands) and used to generate an auto-focus model. During subsequent scans, the auto-focus model can be used to quickly estimate the focal plane of interest for each color channel (or filter band) rather than re-employing the non-synthetic and/or synthetic auto-focus techniques.

Techniques for acquiring focused images of a microscope slide are disclosed. During a calibration phase, a “base” focal plane is determined using non-synthetic and/or synthetic auto-focus techniques. Furthermore, offset planes are determined for color channels (or filter bands) and used to generate an auto-focus model. During subsequent scans, the auto-focus model can be used to quickly estimate the focal plane of interest for each color channel (or filter band) rather than re-employing the non-synthetic and/or synthetic auto-focus techniques.

Methods and apparatuses are disclosed whereby structured illumination microscopy (SIM) is applied to a scanning microscope, such as a confocal laser scanning microscope or sample scanning microscope, in order to improve spatial resolution. Particular aspects of the disclosure relate to the discovery of important advances in the ability to (i) increase light throughput to the sample, thereby increasing the signal/noise ratio and/or decreasing exposure time, as well as (ii) decrease the number of raw images to be processed, thereby decreasing image acquisition time. Both effects give rise to significant improvements in overall performance, to the benefit of users of scanning microscopy.