Apparatus and methods for fluorescence imaging using radiofrequency multiplexed excitation. One apparatus splits an excitation laser beam into two arms of a Mach-Zehnder interferometer. The light in the first beam is frequency shifted by an acousto-optic deflector, which is driven by a phase-engineered radiofrequency comb designed to minimize peak-to-average power ratio. This RF comb generates multiple deflected optical beams possessing a range of output angles and frequency shifts. The second beam is shifted in frequency using an acousto-optic frequency shifter. After combining at a second beam splitter, the two beams are focused to a line on the sample using a conventional laser scanning microscope lens system. The acousto-optic deflectors frequency-encode the simultaneous excitation of an entire row of pixels, which enables detection and de-multiplexing of fluorescence images using a single photomultiplier tube and digital phase-coherent signal recovery techniques.

Among other things, motility of at least one individual microorganism or a change in motility of at least one individual microorganism or both is or are characterized. The characterized motility or change in motility is used to detect the presence or count of the at least one individual microorganism, or determine the identity of a species or strain of the at least one individual microorganism, or determine a susceptibility of the at least one individual microorganism to one or more antibiotics or other antimicrobials.

A SPIM-microscope (Selective Plane Imaging Microscopy) and a method of operating the same having a y-direction illumination light source and a z-direction detection light camera. An x-scanner generates a sequential light sheet by scanning the illumination light beam in the x-direction. An electronic zoom is provided that is adapted to change the scanning length in the x-direction independently of a focal length of the illumination light beam and a size of the light sheet in the y-direction and in the z-direction, wherein the number of image pixels in x-direction is maintained unchanged by the electronic zoom independently of the scanning length in x-direction that has been selected.

Provided is a light detection device having a laser light source, a splitting unit, a first modulation unit, a second modulation unit, a first detection unit and a second detection unit that detect light, and a control unit.

A re-scan microscope for forming an image of a sample is disclosed. The system comprises an illumination optical system for directing, and optionally focusing, illumination light at the sample herewith providing an illumination light spot at the sample. The illumination light spot causes emission light from the sample. The microscope system further comprises a detection optical system for focusing at least part of the emission light onto an imaging plane of an imaging system herewith causing an emission light spot on the imaging plane. The microscope system also comprises a rotatable element for, when rotating, moving the illumination light spot over and/or through the sample and simultaneously moving the emission light spot over said imaging plane of the imaging system. The rotatable element comprises at least two reflective surfaces.

A photon peak event detection system accepts an analog output from a photon sensor, directly digitizes the analogy output and includes a graphics processing unit (GPU) programmed to conduct a photon peak event detection in real-time via a photon count program that analyzes the digitized photon sensor output in sampling periods each having at least three consecutive data points to determine a local maximum among the consecutive data points and compare the local maximum to one or more predetermined thresholds to determine whether or not a photon was received in each sampling period, the algorithm providing photon counts to a phasor analysis program in the GPU. The phasor analysis program calculates pixelwise fluorescence lifetime phasor data in real-time and sends the data to a central processing unit.

An object of the present invention is to provide an image processing apparatus, an image processing program, and an image processing method capable of specifying and correcting desired motion. An image processing apparatus includes: global motion estimation means for estimating global motion indicating motion of a specific region containing a specific object in a moving picture from a plurality of frame images contained in the moving picture; local motion estimation means for estimating local motion indicating motion of the specific object in the moving picture from the plurality of frame images; and image correction means for correcting the motion of the specific object or the specific region in the moving picture on the basis of the estimated global motion or the local motion.

For correcting aberration-induced imaging errors of an optical system which includes an objective (14) and an adaptive optic (18), light (5) and a sample (20) are selected such that the light (5), in acting upon the sample (20), reduces a measurement signal (28) from the sample (20), wherein a relative variation of the measurement signal (28) depends on the intensity of the light (5). The measurement signal (28) from a focal area of the optical system in the sample (20) is registered over a first and a later second period of time (38, 37) to determine a first measurement value and a second measurement value. Over a third period of time (39) which overlaps with the first and/or the second period of time, the light (5) is focused into the focal area by means of the optical system. A measure value for the relative variation of the measurement signal (28) is determined from the first and the second measurement values and used in controlling the adaptive optic (18) as a metric to be optimized.

An optical microscope device includes: a first illumination optical system including a light source that emits illumination light for illuminating a specimen an LCOS spatial light modulation element that controls a polarization state of the illumination light, a first illumination optical member that uniformly illuminates the LCOS spatial light modulation element, and a polarization optical element that controls a transmission state of the illumination light directed to the specimen from the LCOS spatial light modulation element in response to the polarization state of the illumination light; a second illumination optical system including a second illumination optical member that images a light flux from the first illumination optical system on a specimen surface; and an imaging optical system for imaging the specimen surface.

A SPIM-microscope (Selective Plane Imaging Microscope) having a y-direction illumination light source and a z-direction detection light camera. An x-scanner generates a sequential light sheet by scanning the illumination light beam in the x-direction. The SPIM-microscope has an illumination optics having a zoom optics provided in a beam path of the illumination light beam, the zoom optics being adapted to change the focal length of the illumination light beam and adapted to detect a larger area of the object by sequentially detecting sequences of images along the y-direction that have an increased resolution along the z-direction. An image processing unit combines these sequences of images by image stitching into one large overall image.