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.

Irradiation light in a visible light region is irradiated to a sample while switching irradiation of infrared light IR having a wavelength that corresponds to the infrared absorption spectrum of an observation target material included in the sample between a first state and a second state. A first image and a second image are generated based on the phase distribution, the intensity distribution, and the polarization direction distribution of the light including the irradiation light that has passed through the sample in synchronization with the switching of the infrared light IR irradiation between the first state and the second state. Subsequently, an output image is generated so as to represent one from among the position, size, and shape based on the difference and/or ratio with respect to the pixel values for each pixel between the first image and the second image.

A system for quantitative differential phase contrast microscopy with isotropic transfer function utilizes a modulation mechanism to create a detection light field having a radial or other axial orientation of optical intensity gradient or other distribution. A condenser generates an off-axis light field to project onto an object under examination, thereby generating an object light field, which is then guided to an image capturing device through an objective lens for capturing images. A differential phase contrast algorithm is applied to the images for obtaining a phase, thereby a depth information corresponding to the phase can be obtained to reconstruct the surface profile of the object.

A system for quantitative differential phase contrast microscopy with isotropic transfer function utilizes a modulation mechanism to create a detection light field having a radial or other axial orientation of optical intensity gradient or other distribution. A condenser generates an off-axis light field to project onto an object under examination, thereby generating an object light field, which is then guided to an image capturing device through an objective lens for capturing images. A differential phase contrast algorithm is applied to the images for obtaining a phase, thereby a depth information corresponding to the phase can be obtained to reconstruct the surface profile of the object.

The present disclosure provides improved systems and methods for automated cell identification/classification. More particularly, the present disclosure provides advantageous systems and methods for automated cell identification/classification using shearing interferometry with a digital holographic microscope. The present disclosure provides for a compact, low-cost, and field-portable 3D printed system for automatic cell identification/classification using a common path shearing interferometry with digital holographic microscopy. This system has demonstrated good results for sickle cell disease identification with human blood cells. The present disclosure provides that a robust, low cost cell identification/classification system based on shearing interferometry can be used for accurate cell identification. For example, by combining both the static features of the cell along with information on the cell motility, classification can be performed to determine the type of cell present in addition to the state of the cell (e.g., diseased vs. healthy).

The present disclosure provides improved systems and methods for automated cell identification/classification. More particularly, the present disclosure provides advantageous systems and methods for automated cell identification/classification using shearing interferometry with a digital holographic microscope. The present disclosure provides for a compact, low-cost, and field-portable 3D printed system for automatic cell identification/classification using a common path shearing interferometry with digital holographic microscopy. This system has demonstrated good results for sickle cell disease identification with human blood cells. The present disclosure provides that a robust, low cost cell identification/classification system based on shearing interferometry can be used for accurate cell identification. For example, by combining both the static features of the cell along with information on the cell motility, classification can be performed to determine the type of cell present in addition to the state of the cell (e.g., diseased vs. healthy).

A high spatial and temporal resolution synthetic aperture phase microscopy (HISTR-SAPM) system and methods are provided for sample imaging and metrology. The HISTR-SAPM system includes a sample-illumination path along which a first illumination beam propagates and a reference-beam path along which a second illumination beam propagates. A first digital micromirror device (DMD), a second DMD, and a first scanning objective lens are disposed in the sample-illumination path and at a first side adjacent to the sample. A second scanning objective lens passes the sample information to a beam splitter (BS), where the sample illumination beam and the reference-beam are combined to form an interferogram at a final image plane for imaging the sample. A Fourier spatial spectrum analysis and a synthetic aperture are then used to reconstruct a quantitative phase map of the sample with a high resolution and at a high-speed.

A microscopic transmitted light contrasting method includes illuminating a sample through asymmetrical first and second illumination pupils and imaging the sample through asymmetrical first and second detection pupil in order to generate, respectively, first and second partial images. The first illumination pupil and the first detection pupil, as well as the second illumination pupil and the second detection pupil, are arranged pivoted in relation to one another and partially overlapping in projection on a plane perpendicular to an optical axis in such a way that first and third regions of an angular space are in a bright field and second and fourth regions of the angular space are in a dark field, and the first and second partial images each have a bright and a dark field component. An image of the sample is generated from the first and second partial images.

A microscopic transmitted light contrasting method includes illuminating a sample through asymmetrical first and second illumination pupils and imaging the sample through asymmetrical first and second detection pupil in order to generate, respectively, first and second partial images. The first illumination pupil and the first detection pupil, as well as the second illumination pupil and the second detection pupil, are arranged pivoted in relation to one another and partially overlapping in projection on a plane perpendicular to an optical axis in such a way that first and third regions of an angular space are in a bright field and second and fourth regions of the angular space are in a dark field, and the first and second partial images each have a bright and a dark field component. An image of the sample is generated from the first and second partial images.

The technology disclosed present systems and methods to produce an enhanced resolution image from images of a target using structured illumination microscopy (SIM). The method includes transforming at least three images of the target captured by a sensor in a spatial domain into a Fourier domain to produce at least three frequency domain matrices that each include first blocks of complex coefficients and redundant second blocks of complex coefficients that are conjugates to the first blocks. The method includes reducing computing resources required to produce the enhanced resolution image by using first blocks of complex coefficients to produce at least three phase-separated half-matrices in the Fourier domain. The method includes performing one or more intermediate transformation on the phase-separated half-matrices to produce realigned shifted half-matrices. The method includes calculating complex coefficients of second blocks in the Fourier domain to produce full matrices from half-matrices.