An in-line holographic microscope can be used to analyze on a frame-by-frame basis a video stream to track individual colloidal particles' three-dimensional motions. The system and method can provide real time nanometer resolution, and simultaneously measure particle sizes and refractive indexes. Through a combination of applying a combination of Lorenz-Mie analysis with selected hardware and software methods, this analysis can be carried out in near real time. An efficient particle identification methodology automates initial position estimation with sufficient accuracy to enable unattended holographic tracking and characterization.

A lighting device for an imaging optical device such as a microscope is provided. The lighting device illuminates an object to be analyzed in an imaging optical device for microscopic analysis in at least two different contrasting techniques. The lighting device has light sources for the illumination, where the light sources are associated with a contrasting technique are controllable independently from each other.

Systems, methods, and apparatus for differential phase contrast microscopy by transobjective differential epi-detection of forward scattered light are provided. In some embodiments, a microscope objective comprises: a housing with mounting threads at a second end; optical components defining an optical axis, comprising: an objective lens mounted at a first end, configured to collect light from a sample placed in a field of view, the plurality of optical components create a pupil plane at a first distance along the optical axis at which rays having the same angle of incidence on the objective lens converge at the same radial distance from the optical axis; a photodetector within the housing offset from the optical axis at a second distance along the optical axis; and another photodetector within the housing at second distance along the optical axis and offset from the optical axis in the opposite direction from the first photodetector.

A confocal optical system includes a light source and a spinning polarizer disposed in the optical pathway such the light emitted from the light source passes through the spinning polarizer. A first objective lens is disposed in the optical pathway to allow passage of light that passes through the spinning polarizer. A microlens array member is disposed adjacent the first objective lens to receive light. The microlens array member includes a plate having a plurality of holes arranged in an array pattern. A second objective lens is disposed in the optical pathway to receive and allow passage of light to a sample. The optical pathway is arranged such that, after reaching the sample, the light is directed back through the second objective lens, the microlens or microlens with filter array, and the first objective lens and a fluorescent filter cube as an emission beam to reach at least one camera which provides an image of the sample.

The invention relates to an optical assembly for scanning excitation radiation and/or manipulation radiation in a laser scanning microscope. The optical assembly according to the invention is characterized in that in addition to a first and a second focusing device, a third focusing device is provided in order to generate a third pupil plane which is optically conjugated to a first pupil plane, a third beam deflecting device is arranged on the third pupil plane in order to deflect the excitation radiation and/or manipulation radiation, a first beam deflecting means is provided between the second focusing device and the second pupil plane and the second pupil plane and the third focusing device in order to deflect the excitation radiation and/or manipulation radiation coming from the third focusing device while bypassing the second beam deflecting device in the direction of the second focusing device, a fourth focusing device is provided for generating a fourth pupil plane which is optically conjugated to the third pupil plane, and a variable second beam deflecting means is arranged on the fourth pupil plane in order to switch an optical beam path between a first beam path and a second beam path. The invention additionally relates to a laser scanning microscope.

Upstream a microscope objective lens, a polarization direction of a light beam is tilted with a first electro-optical deflector between a first polarization direction with which the light beam is deflected by a first polarization beam splitter by a first angle and a second polarization direction with which it is deflected by a second angle. With a second electro-optical deflector, the polarization direction of the light beam is tilted between a third polarization direction with which the light beam is deflected by a second polarization beam splitter by a third angle and a fourth polarization direction with which it is deflected by a fourth angle. By rotating the polarization direction of the light beam by means of the first and second electro-optical deflectors in a coordinated way the light beam is tilted about a fixed point in a pupil of the objective lens.

A method for imaging a sample using a light-sheet microscope includes illuminating the sample from two different illumination directions using two light sheets, which have different polarization states and are superimposed on one another in a coplanar manner in a target region of the sample. An image of the illuminated target region is generated using an imaging optical unit of the light-sheet microscope. An interference pattern is generated using the two light sheets in the illuminated target region, whereby an image modulation corresponding to the interference pattern is applied to the image of the target region. The image modulation is evaluated. The illuminated target region is aligned in dependence on the evaluated image modulation in relation to a focal region of the imaging optical unit.

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 method for imaging a sample, wherein the sample changes a polarization state of light as a function of position, wherein the method includes changing a polarization state of a purely polarized light of an incident light striking a micro-retarder array, thereby inducing a changed polarization state of the polarization state. The micro-retarder array is placed in a rear conjugate focal plane of a microscope. The method additionally includes projecting the changed polarization state of the polarization state into an object plane of the microscope containing the sample.

An optical apparatus for examining a sample includes: an illumination unit for emitting illumination light in an illumination wavelength range onto the sample; a detection unit for collecting detection light in a detection wavelength range from the sample, the illumination wavelength range and the detection wavelength range partially overlapping in an intermediate wavelength range; and a light separating device for separating the illumination light and the detection light, the light separating device including a beam splitter having: a first splitting characteristic with one of transmitting and reflecting light of at least a first polarization state in the illumination wavelength range excluding the intermediate wavelength range; and a polarization-dependent second splitting characteristic with the one of transmitting and reflecting light of the first polarization state and the other of transmitting and reflecting light of a second polarization state in the intermediate wavelength range.