The present invention provides a femtosecond pulse laser modulator and a miniature two-photon microscopic imaging device. The femtosecond pulse laser modulator includes a negative dispersion light path and a positive dispersion light path, wherein the negative dispersion light path includes a laser input fiber, which is configured to transmit laser compensated for through prechirp; the positive dispersion light path is located between a femtosecond pulse laser and the negative dispersion light path, and is configured to compensate for, through prechirp, negative dispersion of the laser produced in the laser input fiber. The femtosecond laser modulator provided in the present invention provides distortion-free transmission for the femtosecond pulse laser with a center wavelength in 920 nm, and effectively excites commonly used biological indicators. The miniature two-photon microscopic imaging device including this femtosecond laser modulator has high speeds in test and application, with high resolution, and is capable of solving the problem of imaging single dendritic spines in a freely behaving animal.

A system for generating a series of reducing intensity laser pulses from an ultrafast pulsed laser source. The reducing intensity series of pulses is equally temporally spaced between pulses from the laser source. The repeating series of reducing intensity laser pulses is fed to a microscope for imaging. The microscope is capable of detecting the fluorescence from a sample generated by each pulse in the series. The data is processed using knowledge of the pulse intensity, location on the test sample, and amount of fluorescence measured to create an increased dynamic range of the image relative to what can be obtained in a normal two-photon imaging system.

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.

An observation apparatus includes: alight source that emits pulsed light; and an objective that includes a first optical element serving as a light guide part and irradiates a sample with the pulsed light. The first optical element consists of a medium that satisfies the following conditional expression for 1 and 2:

0.75<2/1<1.33 where 1=(n2n1)/(21) and 2=(n3n2)/(32) are satisfied (1 indicates a light wavelength of 706.52 nm; 2 indicates a light wavelength of 1529.6 nm; 3 indicates a light wavelength of 2325.4 nm; and n1, n2, and n3 respectively indicate refractive indexes that the medium has for 1, 2, and 3).

An image acquisition device includes a spatial light modulator modulating irradiation light, a control unit controlling a modulating pattern so that a plurality of light converging points are formed in an observation object, a light converging optical system converging the modulated irradiation light, a scanning unit scanning positions of the plurality of light converging points in the observation object in a scanning direction intersecting an optical axis of the light converging optical system, and a photodetector configured to detect a plurality of observation lights generated from the plurality of light converging points. The control unit sets a center spacing between adjacent light converging points on the basis of the positions of the plurality of light converging points in a direction of the optical axis.

An adapter configured to be optically coupled to a plurality of microscopes. The adapter includes 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.

The present disclosure relates to a method for three-dimensional imaging, including introducing upconverting nanoparticles into a sample, illuminating near-infrared laser such that upconverting nanoparticles introduced into a sample is excited, detecting a visible ray emitted from the excited upconverting nanoparticles and capturing and acquiring two-dimensional images by scanning the sample in a depth direction of the sample, and generating a three-dimensional image of the sample using the two-dimensional images.

The present disclosure relates to a method for three-dimensional imaging, including introducing upconverting nanoparticles into a sample, illuminating near-infrared laser such that upconverting nanoparticles introduced into a sample is excited, detecting a visible ray emitted from the excited upconverting nanoparticles and capturing and acquiring two-dimensional images by scanning the sample in a depth direction of the sample, and generating a three-dimensional image of the sample using the two-dimensional images.

Method for determining the characteristics of a system for generating at least one pattern of light, the method comprising: a) providing a desired pattern of light, b) expressing the amplitude and the phase of the output pulse of the system as a function of the input laser pulse and in function of the characteristics of the system to obtain a calculated output pulse, the input laser pulse having a duration below or equal to 1 nanosecond, c) determining at least one characteristic of the system by minimizing a distance between the calculated output pulse and the desired output laser pulse.

Devices and methods for recording dynamics of cellular and/or biochemical processes, including a device including one or more dispersive elements configured to receive a pulsed laser beam with a spectrum of different wavelengths and disperse the spectrum of the pulsed laser beam; and one or more first elements configured to receive the dispersed spectrum of the pulsed laser beam, and generate a multiphoton excitation area in a biological sample by re-overlapping in time and space the dispersed spectrum of the pulsed laser beam on an area in the biological sample, wherein the device is configured to record at high speed changes of cellular and biochemical processes of a population of cells of the biological sample based on generation of the multiphoton excitation area in the biological sample.