Provided is a determination method capable of non-destructively and simply determining a state of a sphere that is an aggregate of a plurality of cells. A phase difference image of a sphere that is an aggregate of a plurality of cells is generated from a hologram obtained by imaging the sphere, and a state of the sphere is determined on the basis of the phase difference image and a shape index value corresponding to a shape of the sphere.

The invention is a method for estimating a representative volume of particles of interest (10 i) immersed in a sample, the sample extending in at least one plane, referred to as the sample plane (P 10), the sample comprising a sphering agent, capable of modifying the shape of the particles, the method comprising the following steps: a) illuminating the sample by means of a light source (11), the light source emitting an incident light wave (12) propagating towards the sample (10) along a propagation axis (Z); b) acquiring, by means of an image sensor (16), an image (I 0) of the sample (10), formed in a detection plane (P 0), the sample being arranged between the light source (11) and the image sensor (16), each image being representative of a light wave (14) referred to as an exposure light wave, to which the image sensor (16) is exposed under the effect of illumination; c) using the image of the sample (I 0), acquired during step b), and a holographic propagation operator, to calculate a complex expression (A (x, y, z)) of the exposure light wave (14) in different positions relative to the detection plane; the method comprising a step of estimating the representative volume (AA) of the particles of interest (10 i) depending on the complex expressions calculated during step c).

Methods and systems for generating a high-color-fidelity and high-resolution color image of a sample are disclosed; which fuses or merges a holographic image acquired at a single wavelength with a color-calibrated image taken by a low-magnification lens-based microscope using a wavelet transform based colorization method. A holographic microscope is used to obtain holographic images which are used to computationally reconstruct a high-resolution mono-color holographic image of the sample. A lens-based microscope is used to obtain low resolution color images. A discrete wavelet transform (DWT) is used to generate a final image that merges the low-resolution components from the lens-based color image and the high-resolution components from the high-resolution mono-color holographic image.

A method for analyzing microorganisms arranged in a sample is provided, the sample including a viability marker to modify an optical property of the microorganisms in different ways depending on whether they are dead or alive, the method including illumination of the sample and acquisition of an image of the latter by an image sensor, the image sensor then being exposed to an exposure light wave; determining positions of different microorganisms from the acquired image; applying a propagation operator to calculate at least one characteristic value of the exposure light wave at each radial position and at a plurality of distances from the detection plane representing a change in the characteristic value between the image sensor and the sample; and identifying living microorganisms according to each profile.

An apparatus for in-line holographic imaging is disclosed. In one aspect, the apparatus includes at least a first light source and a second light source arranged for illuminating an object arranged in the apparatus with a light beam. The apparatus also includes an image sensor arranged to detect at least a first and a second interference pattern, wherein the first interference pattern is formed when the object is illuminated by the first light source and the second interference pattern is formed when the object is illuminated by the second light source. The first and second interference patterns are formed by diffracted light, being scattered by the object, and undiffracted light of the light beam. The at least first and second light sources are arranged at different angles in relation to the object, and possibly illuminate the object using different wavelengths.

An optical apparatus is comprising an optical imaging arrangement generating either an image of the original object field or the field of the original sample at the pupil plane which consist of both amplitude and phase information. The apparatus is further comprising a digital adaptive optics arrangement with a wave front sensor and a computing unit, which is adapted to generate at least one orthogonally translated digital copy of the original sample object field at the spatial Fourier or pupil plane and to analytically calculate a wave front error based on the phase difference between the original sample wave front and its digital copy or copies.

The present invention includes method and device for label-free holographic screening and enumeration of tumor cells in blood for use in connection with cancer treatments and monitoring.

A method for determining a parameter of a particle present in a sample, the method comprising the following steps: a) illuminating the sample with the light source, the light source emitting an incident light wave that propagates along a propagation axis; b) acquiring an image of the sample with the image sensor, the image sensor being exposed to an exposure light wave; c) determining a position of the particle in the detection plane; d) on the basis of the acquired image, applying a propagation operator, for a plurality of distances from a detection plane, so as to estimate, at each distance, a complex amplitude of the exposure light wave; e) on the basis of the complex amplitude estimated, at various distances, obtaining a profile representing a variation of the complex amplitude of the exposure light wave along an axis parallel to the propagation axis and passing through the position of the particle.

The particle may associated with a set of parameters, comprising at least a size of the particle and a refractive index of the particle.

A system for detecting objects in a specimen includes a chamber for holding at least a portion of the specimen. The system also includes a lens-free image sensor for obtaining a holographic image of the portion of the specimen in the chamber. The system further includes a processor in communication with the image sensor, the processor programmed to obtain a holographic image having one or more objects depicted therein. The processor is further programmed to obtain at least one object template representing the object to be detected, and to detect at least one object in the holographic image.

A system for lens-free imaging includes a processor in communication with a lens-free image sensor. The processor is programmed to operate the image sensor to obtain a hologram ??. The processor is further programmed to generate, from the hologram, a reconstructed image X and phase W at a focal depth z using an assumption of sparsity.