Nanjing University of Science and Technology

Jiasong Sun

Qian Chen

Chao Zuo

Shijie Feng

Yuzhen Zhang

Guohua Gu

Yao Fan

Jiaji Li

Liang Zhang

Yan Hu

Wei Yin

Tianyang Tao

Jiaming Qian

Xiangpeng Pan

Jialin Zhang

A single-shot differential phase contrast quantitative phase imaging method based on color multiplexing illumination. A color multiplexing illumination solution is used to realize single-shot differential phase contrast quantitative phase imaging. In the single-shot color multiplexing illumination solution, three illumination wavelengths of red, green, and blue are used to simultaneously illuminate a sample, and the information of the sample in multiple directions is converted into intensity information on different channels of a color image. By performing channel separation on this color image, the information about the sample at different spatial frequencies can be obtained. Such a color multiplexing illumination solution requires only one acquired image, thus enhancing the transfer response of the phase transfer function of single-shot differential phase contrast imaging in the entire frequency range, and achieving real-time dynamic quantitative phase imaging with a high contrast, a high resolution, and a high stability. In addition, an alternate illumination strategy is provided, so that a completely isotropic imaging resolution at the limit acquisition speed of the camera can be achieved.

Single-shot differential phase contrast quantitative phase imaging method based on color multiplexed illumination

A calibration method for fringe projection systems based on plane mirrors. Firstly, two mirrors are placed behind the tested object. Through the reflection of mirrors, the camera can image the measured object from the front and other two perspectives, so as to obtain 360-degree two-dimensional information of the measured object. The projector projects three sets of phase-shifting fringe patterns with frequencies of 1, 8, and 64. The camera captures the fringe image to obtain an absolute phase map with a frequency of 64 by using the phase-shifting method and the temporal phase unwrapping algorithm. By using the calibration parameters between the projector and the camera, the absolute phase map can be converted into three-dimensional information of the measured object. Then, the mirror calibration is realized by capturing a set of 3D feature point pairs, so that the 3D information from different perspectives is transformed into a unified world coordinate system. The calibration method does not need to artificially fix the feature pattern on plane mirrors, only needs to capture a set of 3D feature point pairs by the camera to directly realize the mirror calibration that it avoids the loss of measurement accuracy and realizes high-precision panoramic three-dimensional measurement.

Calibration method for fringe projection systems based on plane mirrors

The present invention discloses a three-dimensional diffraction tomography microscopy imaging method based on LED array coded illumination. Firstly, acquiring the raw intensity images, three sets of intensity image stacks are acquired at different out-of-focus positions by moving the stage or using electrically tunable lens. And then, after acquiring the intensity image stacks of the object to be measured at different out-of-focus positions, the three-dimensional phase transfer function of the microscopy imaging system with arbitrary shape illumination is derived. Further, the three-dimensional phase transfer function of the microscopic system under circular and annular illumination with different coherence coefficients is obtained as well, and the three-dimensional quantitative refractive index is reconstructed by inverse Fourier transform of the three-dimensional scattering potential function. The scattering potential function is converted into the refractive index distribution. Thus, the quantitative three-dimensional refractive index distribution of the test object is obtained. The invention realizes high-resolution and high signal-to-noise ratio 3D diffraction tomography microscopic imaging of cells, tiny biological tissues and other samples.

3D diffraction tomography microscopy imaging method based on LED array coded illumination

A microscopic imaging method of phase contrast (PC) and differential interference contrast (DIC) based on the transport of intensity equation (TIE) includes capturing three intensity images along the optical axis; solving the TIE by deconvolution to obtain the quantitative phase; obtaining the intensity image under the DIC imaging mode according to the DIC imaging principle; and obtaining the corresponding phase image of PC imaging mode according to the PC imaging principle. The method can endow the bright-field microscope with the ability to realize PC and DIC imaging without complex modification of the traditional bright-field microscope. In addition, it has the same imaging performance as the phase contrast microscope and differential interference contrast microscope, which are expensive, complex-structure, and has strict environmental conditions.

Microscopic imaging method of phase contrast and differential interference contrast based on the transport of intensity equation

The invention discloses a programmable annular LED illumination-based high efficiency quantitative phase microscopy imaging method, the proposed method comprising the following steps: the derivation of system optical transfer function in a partially coherent illumination imaging system; the derivation of phase transfer function with the weak object approximation under the illumination of tilted axially symmetric coherent point illumination source; the extension of illumination from an axially symmetric coherence point source to a discrete annular point source, and the optical transfer function can be treated as an incoherent superposition of each pair of tilted axially symmetric coherent point sources. The acquisition of raw intensity dataset; the implementation of deconvolution for quantitative phase reconstruction. The invention derives the system phase transfer function under the tilted axially symmetric point light source in the case of partially coherent illumination, and promotes the optical phase transfer function of the discrete annular point light source. The programmability characteristic of LED array enables the annular illumination aperture to be flexibly adjustable, being applicable to different microscopic objects with different numerical apertures, and improving the compatibility and flexibility of the system.

Programmable annular led illumination-based high efficiency quantitative phase microscopy imaging method

The patent discloses a differential phase contrast (DPC) quantitative phase microscopy method based on the optimal illumination pattern design. Firstly, the optimal illumination pattern corresponding to the isotropic phase transfer function of DPC quantitative phase imaging is derived, which is determined as a semi-annular illumination pattern with the illumination numerical aperture NAequal to the numerical aperture NAof the objective lens. The illumination intensity distribution varies with the cosine of the illumination angle, and it can be expressed as S(&#x3b8;)=cos(&#x3b8;). This patent effectively compensates for the frequency loss of phase transfer, not only the high-frequency responses of PTF are enhanced, but also the transfer responses of low-frequency phase information is significantly improved. As a result, the optimal illumination scheme ensures the correctness and achieves high resolution phase reconstruction, while the number of illuminations is reduced to a minimum of two, which greatly increases the imaging speed, allowing for real-time dynamic, high-correctness, high-resolution phase imaging results.

Quantitative phase imaging method based on differential phase contrast with optimal lighting pattern design

A high-illumination numerical aperture-based large field-of-view high-resolution microimaging device, and a method for iterative reconstruction, the device comprising an LED array (), a stage (), a condenser (), a microscopic objective (), a tube lens (), and a camera (), the LED array () being arranged on the forward focal plane of the condenser (). Light emitted by the i-th lit LED unit () of the LED array () passes through the condenser () and converges to become parallel light illuminating a specimen () to be examined, which is placed on the stage (); part of the diffracted light passing through the specimen () is collected by the microscopic objective (), converged by the tube lens (), and reaches the imaging plane of the camera (), forming an intensity image recorded by the camera (). The present device and method ensure controllable programming of the illumination direction, while also ensuring an illumination-numerical-aperture up to 1.20 and thus achieving a reconstruction resolution up to 0.15 &#x3bc;m.

High-illumination numerical aperture-based large field-of-view high-resolution microimaging device and a method for iterative reconstruction

An annular-irradiation high-resolution quantitative phase microimaging based on light intensity transfer equation is proposed here includes designing an annular aperture for the imaging system illumination; invoking the weak object approximation by using the parameters of annular illumination aperture and bright field microscopy to calculate a weak object optical transfer function (WOTF) on the basis of a partially coherent imaging theory; and collecting three intensity images by a camera and obtaining the quantitative phase image of object by resolving the light intensity transfer equation with a deconvolution algorithm.

Annular-irradiation high-resolution quantitative phase microimaging method using an annular illumination aperture

A super-rapid three-dimensional measurement method and system based on an improved Fourier transform contour technique is disclosed. The method comprises: firstly calibrating a measurement system to obtain calibration parameters, then cyclically projecting 2n patterns into a measured scene using a projector, wherein n patterns are binary sinusoidal fringes with different high frequency, and the other n patterns are all-white images with the values of 1, and projecting the all-white images between every two binary high-frequency sinusoidal fringes, and synchronously acquiring images using a camera; and then performing phase unwrapping on wrapped phases to obtain initial absolute phases, and correcting the initial absolute phases, and finally reconstructing a three-dimensional topography of the measured scene by exploiting the corrected absolute phases and the calibration parameters to obtain 3D spatial coordinates of the measured scene in a world coordinate system, thereby accomplishing three-dimensional topography measurement of an object. In this way, the precision of three-dimensional topography measurement is ensured, and the speed of three-dimensional topography measurement is improved.

Super-rapid three-dimensional topography measurement method and system based on improved fourier transform contour technique

A highly efficient three-dimensional image acquisition method based on multi-mode composite encoding and epipolar constraint, respectively using a fast imaging mode or a high-precision imaging mode, wherein in the fast imaging mode, two phase maps having different frequencies are obtained by four stripe gratings, and a high-frequency absolute phase is obtained by means of the epipolar constraint and a left-right consistency check, and the three-dimensional image is obtained by means of a mapping relationship between the phase and three-dimensional coordinates; and in the high precision imaging mode, two phases having different frequencies are obtained by means of N+2 stripe gratings, a low-frequency absolute phase is obtained by the epipolar constraint, and the unwrapping of a high-frequency phase is assisted by means of the low-frequency absolute phase, so as to obtain the high-frequency absolute phase, and finally, the three-dimensional image is obtained by the mapping relationship between the phase and the three-dimensional coordinates. In this way, the imaging efficiency is ensured, and the imaging precision is improved.

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Nanjing, China

Jiasong Sun