A processor for demixing a fluorescent-light input signal of a fluorescence microscope, the fluorescent-light input signal including at least two fluorescence emission responses that overlap in time, each of the at least two fluorescence emission responses being representative of an individual impulse response of a fluorophore to a fluorescence-triggering light pulse of a clocked time series of fluorescence-triggering light pulses, the processor: receiving a trigger signal comprising a time series of time markers, the trigger signal being representative of a clocking rate, at which the clocked time series of fluorescence-triggering light pulses is generated; and separating at least one fluorescence emission response from the fluorescent-light input signal.

The present invention relates to an inline scanning holography system for a phosphor and a transmitter. According to the present invention, the inline scanning holography system includes a polarization sensitive lens that receives a linearly polarized beam and generates a first spherical wave of right-handed circular polarized light having a negative focal length and a second spherical wave of left-handed circular polarized light having a positive focal length, a polarizer that passes only a beam component in a predetermined polarization direction therethrough among components of the generated first and second spherical waves, a scanning unit for scanning a phosphor by using an interference beam generated between the first and second spherical waves passing through the polarizer, and a first photodetector that detects a fluorescent beam diverged from the phosphor. According to the present invention, a high-efficiency and high-quality optical scanning holography for a phosphor or a transmitter may be implemented.

A fluorescence scanning microscope includes excitation and de-excitation light sources, which are designed to generate an excitation and a de-excitation light distribution, respectively. An illumination unit combines the light distributions to form a light distribution scanning over multiple illumination target points of a sample in such a way that an intensity maximum of the excitation light distribution and an intensity minimum of the de-excitation light distribution are spatially superimposed on one another. A detector detects fluorescence photons emitted from the respective illumination target point as a function of their arrival times. A processor evaluates the fluorescence photons with respect to the arrival times, generates a first pixel and a second pixel based thereon, assembles the first and second pixels to form first and second sample images, respectively, and, by means of the two sample images, determines a spatial offset between the intensity maximum and the intensity minimum.

A method of fluorescence spectroscopy includes providing a high-performance sensor that combines imaging with high intrinsic time resolution and high-rate capability, and resolving fluorescence data in four dimensions.

A scanning microscope includes an objective and a scanning element that is adjustable for a time-variable deflection to guide a focused illumination beam across the sample in a scanning movement. A detection beam is guided across sensor elements of an image sensor in a movement which corresponds to the scanning movement of the focused illumination beam. A dispersive element of a predetermined dispersive effect arranged upstream of the image sensor spatially separates different spectral components of the detection beam from one another on the image sensor. A controller detects the time-variable adjustment of the scanning element, assigns the spatially separated spectral components of the detection beam to the sensor elements of the image sensor based on the detected time-variable adjustment, while taking into account the predetermined dispersive effect of the dispersive element, and individually reads out the sensor elements assigned to the spectral components.

A method of imaging a sample by interferometric scattering microscopy, the method comprising illuminating a sample with at least one light source, the sample being held at a sample location comprising a reflective surface, such that a reflected signal is formed; the reflected signal comprising light from the light source and light scattered by the sample; detecting the output light over a first time window for a first frame N.sub.1; detecting the output light over a second time window for a second frame N.sub.2; calculating a ratiometric signal R which is the ratio of N.sub.1 and N.sub.2 minus 1; estimating the ratiometric motion signature S=(S.sub.x, S.sub.y) from frames N.sub.1 and N.sub.2 with S defined as the ratiometric image which would be measured from an invariant sample moving along x and y for a given motion vector m=(m.sub.x, m.sub.y); estimating m as the most consistent vector such that R is approximated using S and m; calculating the corrected ratiometric contrast image R* from R, S and m.

A coherent anti-stokes Raman scattering apparatus for imaging a sample includes an optical output; an optical source arranged to generate a first optical signal at a first wavelength; and a nonlinear element arranged to receive the first optical signal, where the nonlinear element is arranged to cause the first optical signal to undergo four-wave mixing on transmission through the nonlinear element such that a second optical signal at a second wavelength and a third optical signal at a third wavelength are generated, wherein an optical signal pair including two of the first, second and third optical signals is provided to the optical output for imaging the sample.

An apparatus is configured to count N-photon events within a time-dependent sequence of events of interactions of a plurality of photons with a light sensitive detector. The apparatus includes a signal-processing device and the light sensitive detector. An N-photon event represents an occurrence of at least N timely overlapping single photon events. The light sensitive detector is adapted to generate a time-dependent digital signal comprising digital patterns representing the time-dependent sequence of events from the detection of the plurality of photons with the light sensitive detector. Each digital pattern in the digital signal comprises a digital pattern width having a continuous sequence of digital values representing at least one event of interaction of at least one photon with the light sensitive detector. The signal-processing device is adapted to identify N-photon events from the digital patterns in the digital signal in dependence from the respective digital pattern width.

A microscope comprising an illumination assembly, an image capture device and a processor can be configured to selectively identify regions of a sample comprising artifacts or empty space. By selectively identifying regions of the sample that have artifacts or empty space, the amount of time to generate an image of the sample and resources used to generate the image can be decreased substantially while providing high resolution for appropriate regions of the computational image. The processor can be configured to change the imaging process in response to regions of the sample that comprises artifacts or empty space. The imaging process may comprise a higher resolution process to output higher resolution portions of the computational image for sample regions comprising valid sample material, and a lower resolution process to output lower resolution portions of the computational image for sample regions comprising valid sample material.

A beam-scanning optical design is described for achieving up to kHz frame-rate optical imaging on multiple simultaneous data acquisition channels. In one embodiment, two fast-scan resonant mirrors direct the optical beam on a circuitous trajectory through the field of view, with the trajectory repeat-time given by the least common multiplier of the mirror periods. Dicing the raw time-domain data into sub-trajectories combined with model-based image reconstruction (MBIR) 3D in-painting algorithms allows for effective frame-rates much higher than the repeat time of the Lissajous trajectory. Because sub-trajectory and full-trajectory imaging are different methods of analyzing the same data, both high-frame rate images with relatively low resolution and low frame rate images with high resolution are simultaneously acquired.