A method of investigating a sample using a wide-field multiphoton microscope with tunable excitation wavelength (λ) is disclosed. The sample is illuminated at some excitation wavelength. Images of the sample are acquired at wavelengths different from the excitation wavelength, e.g., at half the excitation wavelength for SHG microscopy, using an image sensor. Based on the obtained images, an autoalignment procedure is carried out for optimizing the position of the sample relative to the illumination and/or collection beam paths. An image or spectrum that has been obtained for the optimal relative position is stored. The procedure is repeated for multiple excitation wavelengths. The autoalignment procedure can comprise an autofocusing subprocedure to automatically optimize the position of the sample relative to a focal plane of at least one objective of the multiphoton microscope along a direction that is perpendicular to the focal plane, and an in-plane repositioning subprocedure for automatically optimizing the position of the sample within the focal plane.

Methods and systems may provide for 3D volumetric displays. Such 3D volumetric displays may include a transparent enclosed volume holding a noble gas as a gain medium. Two electrodes positioned on opposing sides of the transparent enclosed volume, may apply a voltage to excite electrons of the gain medium to an excited state. A pump laser may emit a laser beam into the gain medium at a wavelength that has an energy below an absorption line of the gain medium to allow for photon collision while also allowing the laser beam to enter the gain medium. A lens may focus the laser beam to a focused spot within the transparent enclosed volume and move the focused spot as a three-dimensionally scanned voxel to produce a 3D image.

A multiplexing module provided herein is configured to perform operations of receiving a plurality of laser pulses from a pulsed laser source; splitting each laser pulse into a plurality of beamlets; introducing a delay between each adjacent beamlet of the plurality of beamlets, such that the plurality of beamlets associated with a respective laser pulse of the plurality of laser pulses is distributed equally across a pulse repetition period associated with the pulsed laser source; changing a divergence of each subsequent beamlet of the plurality of beamlets associated with each respective laser pulse to introduce a distinguishing feature between each beamlet of the plurality of beamlet to cause each beamlet to focus on a different axial plane or lateral position of the sample; and outputting the plurality of beamlets associated with each respective laser pulse.

A multi-photon imaging system includes a laser module having a first channel for outputting a two-photon excitation laser pulse and a second channel for outputting a three-photon excitation laser pulse. The system further includes a first optical path for guiding the two-photon laser pulse from the first channel of the laser module and a second optical path for guiding the three-photon laser pulse from the second channel of the laser module. A microscope is also provided for simultaneously receiving the two-photon laser pulse from the first optical path and the three-photon laser pulse from the second optical path, and simultaneously, or with well controllable delays, delivering the two-photon laser pulse and the three-photon pulse to a target volume. The system further includes a photodetector configured to collect photons generated within the target volume in response to simultaneous excitation of the target volume by both the two-photon laser pulse and the three-photon laser pulse.

A microscope directs light through an excitation objective to generate a lattice light sheet (LLS) within a sample. A detection objective collects signal light from the sample in response to the LLS and images the collected light onto a detector. Second and third light beams are imaged onto focal planes of the excitation objective and detection objective, respectively. One or more wavefront detectors determine wavefronts of light emitted from the sample and through the excitation objective in response to the imaged second light beam and emitted from the sample through the detection objective in response to the imaged third light beam. A wavefront of the first light beam is modified to reduce a sample-induced aberration of the LLS within the sample, and a wavefront of the signal light emitted from the sample is modified to reduce a sample-induced aberration of the signal light at the detector.

An optical imaging device for a microscope includes an objective and an optical system configured to interact with the objective for optically imaging an object selectively in a first operating mode and a second operating mode. The optical system includes a first optical subsystem associated with the first operating mode, and a second optical subsystem associated with the second operating mode. The first optical subsystem is configured to form a first image of the object with a first magnification. The second optical subsystem is configured to form a second image of the object with a second magnification that is less than the first magnification. The second optical subsystem includes an optical module insertable into the optical path for selecting the second operating mode. The optical module includes a lens element with a positive refractive power.

A method of generating an image of a sample is provided. The method comprises providing a plurality of photon detectors, scanning the sample with an excitation beam over a predetermined time period, the detectors receiving photons emitted by the sample due to the excitation during the time period. A plurality of intensity images associated with each of the detectors are generated, each being proportional to the mean number of photons detected per unit time. A plurality of correlation images associated with each combination of two of the detectors are generated, each of the correlation images being proportional to the variance of the distribution of detected photons per unit time. The image of the sample is generated using joint sparse recovery from the plurality of intensity and correlation images, wherein the intensity and correlation images have common support.

A system includes a light source to generate an optical signal having a set of pulses at a first repetition rate. The system also includes a multiplexer circuit to generate a multiplexed optical signal from the optical signal n sets of pulses at a second repetition rate, where the n sets of pulses have different polarization states and are at the first repetition rate. The system also includes a focusing unit to split the multiplexed optical signal into n excitation signals to excite a sample. The system also includes an objective to receive the n excitation signals and to illuminate the sample. The objective and the focusing unit collectively focus each excitation signal of the n excitation signals on a different focal plane of the sample to generate a response signal. The system also includes a demultiplexer circuit to generate n emission signals based on the response signal.

Disclosed herein are adapters configured to be optically coupled to a plurality of microscopes, said adapter comprising: a) a first microscope interface configured to optically couple a first microscope to an optical element in optical communication with an optical probe; b) a second microscope interface configured to optically couple a second microscope to the optical element in optical communication with the optical probe; and c) an optical arrangement configured to direct light collected from a sample with aid of the optical probe to (1) the first microscope and second microscope simultaneously, or (2) the first microscope or second microscope selectively.

Methods and systems may provide for 3D volumetric displays. Such 3D volumetric displays may include a transparent enclosed volume holding a noble gas as a gain medium. Two electrodes positioned on opposing sides of the transparent enclosed volume, may apply a voltage to excite electrons of the gain medium to an excited state. A pump laser may emit a laser beam into the gain medium at a wavelength that has an energy below an absorption line of the gain medium to allow for photon collision while also allowing the laser beam to enter the gain medium. A lens may focus the laser beam to a focused spot within the transparent enclosed volume and move the focused spot as a three-dimensionally scanned voxel to produce a 3D image.