An optical scanning microscope includes an illumination system having a light source portion emanating from a light source, first and second polarizing beam splitters, and first and second optical channels disposed between the beam splitters. The light source portion is configured to emit a first illumination light beam comprising light of a first main polarization direction and of a second main polarization direction. The first beam splitter is configured to guide the light primarily into the first and channels, respectively. The second beam splitter is configured to form a second illumination light beam from light of the first and second main polarization directions from the first channel and second channels, respectively. The first and second channels are configured to emit the light of the first and second main polarization directions from the first and second channels, respectively, so as to have different types of convergence.

A tunable filter may include an input focusing optic, an output focusing optic, a linearly-varying filter located at a back focal plane of the input focusing optic and a front focal plane of the output focusing optic, an input angular scanning component located at a front focal plane of the input focusing optic configured to receive an input beam, and an output angular scanning component located at a back focal plane of the output focusing optic. The input focusing optic may receive the input beam from the input angular scanning component and direct the input beam to the linearly-varying filter, where a position of the input beam on the linearly-varying filter is selectable based on an angle of the input angular scanning component. The output focusing optic may receive a filtered beam from the linearly-varying filter and direct the filtered beam to the output angular scanning component.

Systems and methods for three-dimensional fluorescence polarization excitation that generates maps of positions and orientation of fluorescent molecules in three or more dimensions are disclosed.

Methods and apparatus for obtaining a vibrational circular dichroism (VCD) image using a discrete frequency infrared (DFIR) microscope are disclosed. The method includes generating a pulsed laser beam comprising a spectral frequency, which may be tunable; modulating the laser beam to generate circularly polarized light; illuminating a sample and collecting, and detecting an optical signal transmitted or transflected from the location of the sample. The detected signal is demodulated at, for example, both the pulse frequency and the sum or difference of the pulse frequency and the modulating frequency to obtain an intensity value that correspond to the absorbance, and a polarization-dependent value that corresponds to the VCD. Other configurations of the apparatus may be employed to measure VCB and VLD.

Snapshot phase-shifting diffraction modules and associated systems and methods are described that enable high spatial and temporal resolution phase imaging with high immunity to environmental factors such as vibrations and temperature changes. One example optical diffraction phase module includes a polarization grating to produce two circularly polarized light beams with opposite polarizations, a first lens to receive the two circularly polarized beams, and a spatial filter positioned at a focal plane of the first lens. The spatial filter includes two openings, one to spatially filter one of the two circularly polarized light beams, and another opening to allow another circularly polarized light beam to pass. The module also includes a second lens to focus the received light onto an image plane and to enable a phase measurement based a plurality of interferograms. The phase module can be incorporated into a microscope system that operates a reflection or a transmission mode.

Systems and methods for three-dimensional fluorescence polarization excitation that generates maps of positions and orientation of fluorescent molecules in three or more dimensions are disclosed.

The invention relates to a beam manipulation device for a scanning microscope, comprising a main colour splitter for coupling excitation light into an illumination beam path and for separating excitation light from detection light of a detection beam path, said device comprising a scanner, preferably positioned on a pupil plane, for scanning the excitation light. The device is characterised in that: an additional optical section is provided comprising optical elements which influence a beam path; at least one pupil plane and/or at least one intermediate image plane is formed in the additional optical section by the optical elements which influence the beam path; and an adjustable selection device is provided for activating either a first beam segment of the illumination and/or detection beam path, or the additional optical section, wherein the first beam segment of the illumination and/or detection beam path does not contain a pupil plane of the illumination and/or detection beam path.

Techniques to improve image quality in holography utilizing lenses made from materials with non-quantized anisotropic electromagnetic properties, such as birefringent materials, to advantageously split an incoming beam of light into two coincident beams with different focal lengths that interfere with one another and thus create holograms free of electro-optical or pixelated devices are disclosed for microscopy and other applications. The use of thin birefringent lenses and single crystal alpha-BBO lenses are introduced. Corresponding systems, methods and apparatuses are described.

Systems and methods for three-dimensional fluorescence polarization excitation that generates maps of positions and orientation of fluorescent molecules in three or more dimensions are disclosed.

A light microscope comprises: a structuring optical unit comprising a waveguide chip for providing a structured illumination; an input selection device for variably directing light to one of several inputs of the waveguide chip; the waveguide chip further comprising a light guide path following each of the inputs; each light guide path divides into several path divisions; and each path division leads to one output of the wave-guide chip. The outputs of the waveguide chip can be arranged at a pupil plane of the light microscope, and an exit direction of light from the outputs is transverse to a plane defined by the waveguide chip. A method for providing structured illumination light using the light microscope is also described.