81 results about "Phase contrast microscopy" patented technology

The invention provides an apparatus and process for detecting the degree of confluence of animal cells being cultured in a well plate. A well plate is arranged in an imaging station and illuminated with a ring of LEDs, or other optical source, from below at an oblique angle. An image of the well is captured with a CCD camera or other detector from above or below, such that the well image is taken in a dark field configuration where light from the optical source, if not scattered, does not contribute to the well image. By the simple solution of illuminating wells of a well plate from below at an oblique angle, it has been found that many animal cell types can be imaged with sufficient contrast to allow cell identification and consequent cell area computation using image processing techniques, thereby allowing confluence to be determined of animal cells being cultured in well plates. This avoids the need for more complex optical imaging techniques, such as phase contrast microscopy.

A well plate of non-unitary construction. The well plate is constructed from a first part of interconnected tubes that define the side walls of each well and a second part defining the well bases. The hydrophobicity of the first part is selected to suppress meniscus formation in the wells, thereby assisting optical characterization of samples in the wells, in particular by phase contrast microscopy. This can be achieved by using a natural cycloolefin copolymer (COP) which is not subjected to the usual plasma treatment to improve its wetting. The second part is formed of a single transparent sheet of hydrophilic material, such as a COP, which is bonded to the first part, for example ultrasonically.

An optical microscope is provided with an adjustable optical phase ring. The adjustable ring provides a way to compensate for distortion in the visible phase ring before the light reaches the sample. In an inverted microscope, when observing transparent cells under a liquid, the visible light phase ring is distorted. By the use of a Liquid Crystal Display (LCD) in place of a fixed ring, the projected ring is adjusted to realign the light and produce phase. In a typical micro plate, the meniscus formed produces a lens effect that is realigned by providing changes in the position and pattern, to allow phase imaging over a wider portion of the well. The realignment of the ring can be manual or automated and can be dynamically adjusted based upon an observed image of the sample.

A system and method for monitoring a surface or interfacial area. The system and method includes an intense X-ray beam directed to a surface or interface at a low angle to achieve specular reflection with phase contrast associated with an event, such as changing topography, chemistry or magnetic state being detected by a CCD. Upstream or downstream processing can be carried out with the X-ray phase contrast system.

A method of creating a phase contrast image is provided. In some embodiments the method comprises illuminating the target region of a sample with a first light source to provide a first oblique back illumination of the target region of the sample, and detecting a first phase contrast image from light originating from the first light source and back illuminating the target region of the sample. In some embodiments the method further comprises illuminating the sample with a second light source to provide a second oblique back illumination of the target region of the sample, and detecting a second phase contrast image from light originating from the second light source and back illuminating the target region of the sample. In some embodiments a difference image of the target region of the sample is created by subtracting the second phase contrast image of the target region of the sample from the first phase contrast image of the target region of the sample. Apparatus for carrying out the methods are also provided. The methods and apparatus find use, for example, in endoscopy.

A method and objective apparatus are provided for implementing an enhanced phase contrast microscope. A focusing vortex lens, defined by a diffractive spiral zone plate (SZP) lens, is used for the objective for the phase contrast microscope. The SZP lens focuses and imparts a helical phase to incident illumination to image the specimen with spiral phase contrast. The spiral phase contrast microscope is sensitive to phase gradients in all sample axes. Replacing the objective of a microscope with the diffractive SZP lens of the invention immediately provides existing instruments with spiral phase contrast capability.

The invention discloses an automatic stem cell counting method based on depth learning, which relates to the technical field of cell counting. The method comprises the following steps: S11, a phase contrast microscope is adopted to photograph cells in need of counting, and a stem cell image is generated; S12, image preprocessing is carried out, and the cell image is subjected to noise reduction processing and illumination equalization processing to obtain a stem cell image with equalized illumination; S13, a cell artifact generated during the photographing process by the phase contrast microscope is removed; S14, the stem cell image after the artifact is removed is segmented, and multiple candidate stem cell images are acquired; S21, the segmented multiple candidate stem cell images are manually marked, and a training set is built; S22, the training set is inputted to a CNN for training; and S23, a stem cell counting result is counted. The defect that a large amount of manpower is consumed in the traditional manual cell counting can be solved, the defect that a flow counting method damages the cell growth environment is overcome, counting is carried out through the photographed cell image, and the method has the advantages of being stable, efficient, automatic and lossless.

The invention relates to a culture method for efficiently obtaining adipose mesenchymal stem cells, and solves the problem that the cell viability obtained in the prior art is low. The culture method comprises the following steps: separation of the adipose mesenchymal stem cells: washing D.Hanks with fat tissues, detecting to guarantee no pollution, adding a solution mixture of I-type collagenase and trypLETM digestive enzyme with several times of volume, and stopping enzymolysis after carrying out rotary vibration digestion for dozens of minutes at the temperature of 37 DEG C, centrifuging to remove a liquid supernatant, and filtering with screen cloth to obtain an adipose mesenchymal stem cell suspension; culture of the adipose mesenchymal stem cells: inoculating a culture bottle bloodless culture medium with the adipose mesenchymal stem cell suspension, carrying out primary cell culture, observing with an inverted phase contrast microscope in the culture period, and changing a fresh culture medium, when the cells are cultured to 80% to 90% fusion, extensively inoculating a new culture bottle with the cells for culture after digestion with trypLETM, and subculturing for several generations to obtain the purified mesenchymal stem cells. The culture method has the advantages that the digestion is more sufficient, the cell yield is higher, the mesenchymal stem cells are protected from being damaged, and the viability is high.

The invention provides a separating and grouping method of adhesion conditions of main information of cells under a contrast phase microscope for interframe-free reference information. The separating and grouping method of adhesion conditions of main information of cells under a contrast phase microscope for interframe-free reference information includes the following steps: 1) based on analysis of image characteristics of cells under a contrast phase microscope, defining the main information of the images of cells under the contrast phase microscope after constructing hierarchy information in the images; 2) manually marking the key information, referring to the original images and the main information during the marking process, and dividing the adhesion conditions of the main information of the cells into one area, thus being convenient for separately analyzing the conditions; 3) utilizing the separating and grouping method of adhesion conditions of the main information of cells under the contrast phase microscope for interframe-free reference information, separating and grouping the target adhesion conditions; and 4) updating the main information labels and the color marks in the images of the cells under the contrast phase microscope, and obtaining the main information after image restoration. The separating and grouping method of adhesion conditions of main information of cells under a contrast phase microscope for interframe-free reference information can effectively process detection and segmentation of the images of the cells under the contrast phase microscope.

A phase contrast electron microscope has an objective (8) with a back focal plane (10), a first diffraction lens (11), which images the back focal plane (10) of the objective (8) magnified into a diffraction intermediate image plane, a second diffraction lens (15) whose principal plane is mounted in the proximity of the diffraction intermediate image plane and a phase-shifting element (16) which is mounted in or in the proximity of the diffraction intermediate image plane. Also, a phase contrast electron microscope has an objective (8) having a back focal plane (10), a first diffraction lens (11), a first phase-shifting element and a second phase-shifting element which is mounted in or in the proximity of the diffraction intermediate image plane. The first diffraction lens (11) images the back focal plane of the objective magnified into a diffraction intermediate image plane and the first phase-shifting element is mounted in the back focal plane (10) of the objective (8). With the magnified imaging of the diffraction plane by the diffraction lens, the dimensional requirements imposed on the phase plate having the phase-shifting element are reduced.

A single-axis illumination system for a multiple-axis imaging system, particularly an array microscope. A single-axis illumination system is used to trans-illuminate an object viewed with an array of imaging elements having multiple respective axes. The numerical apertures of the imaging elements are preferably matched to the numerical aperture of the illumination system. For Kohler illumination, the light source is placed effectively at the front focal plane of the illumination system. For critical illumination, the light source is effectively imaged onto the object plane of the imaging system. For dark field illumination, an annular light source is effectively provided. For phase contrast microscopy, an annular phase mask is placed effectively at the back focal plane of the objective lens of the imaging system and a corresponding annular amplitude mask is provided effectively at the light source. For Hoffman modulation contrast microscopy, an amplitude mask is placed effectively at the back focal plane of the objective lens of the imaging system and a slit is provided at a source of light of the illumination system. Structured illumination and interferometry, and a secondary source, may also be used with trans-illumination methods and apparatus according to the present invention.

The invention discloses a multi-channel white light common-channel interference microscopic chromatography system. The system comprises a phase contrast microscope and a polarization plate, wherein the polarization plate is arranged on the back focal plane of an objective lens of the phase contrast microscope, the polarization direction of the conjugate surface of the polarization plate is orthogonal with the polarization direction of the complementary surface of the polarization plate, and an airspace phase shift interference module is arranged behind the imaging surface of the phase contrast microscope. The airspace phase shift interference module is composed of a phase adjusting device, a beam splitting lens assembly, a quarter-wave plate (15), a polarization beam splitter and an image sensor. According to the multi-channel white light common-channel interference microscopic chromatography system, only an objective lens with a phase plate needs to be used instead on the basis of an existing phase contrast microscope, real-time measurement can be really achieved, an object can be measured quantitatively, and three-dimensional information of the object can be measured in real time in a lossless mode.

A method for detect bacterial drug resistance by single cell analysis comprises mixing a certain amount of bacterial sample and a certain amount of antibiotic diluent on a slide, covering that slide with a cover glass and sealing with a sealant on four side to form a plate-type microbiochemical reaction system in which the bacterial and antibiotic diluent react biochemically. Bacteria were observed and located by phase contrast microscope and Raman spectra were obtained by laser scanning. Bacterial drug resistance could be obtained by analysis. The invention also provides a device for realizing the method, and the device comprising a plate-type micro-biochemical reaction system, a phase contrast microscope, a high-power optical microscope, a Raman spectrum excitation system and a Raman spectrum collection system. Bacteria were observed by phase contrast microscope and high power optical microscope, and then Raman spectra were obtained by Raman spectrum excitation system and Raman spectrum collection system to detect bacterial drug resistance.

This invention discloses a heat plasticity macromolecule material film or pad detecting sample hot-press and belongs to hot-press apparatus of macrodolecule. The hot-press machine comprises temperature control meter, supportive column, bottom panel, parallel guide column, lift dowel, top panel, and level beam and pressure device. It is characterized by the following: the top and bottom panels comprises the heat preservation cover, top wire, sample protruding table, pad heater, heat isolator, and temperature sensor. The pressure machine comprises pressure plate, pressed bolts, spring cushion pad, spring, and bolts.

The phase contrast microscope includes: an illumination optical system 10 that emits illumination light for phase difference measurement to an observation target placed in a container; an adjustment optical system 20 that is provided between the illumination optical system 10 and the observation target S, has at least one optical element 21, and adjusts refraction of the illumination light due to a liquid surface shape of a liquid C in the container 60; an imaging unit 40 that images the observation target to which the illumination light has been emitted; and an adjustment optical system control unit 51 that adjusts optical characteristics of the adjustment optical system 20 based on uniformity of a density of an image captured by the imaging unit 40 and a density contrast.

Atomic force microscope measuring device comprising a micro-cantilever and an intensity modulated laser exciting the cantilever, wherein the measuring device comprises an optical microscope, in particular a fluorescence microscope, a confocal microscope, a fluorescence energy transfer (FRET) microscope, a DIC and / or phase contrast microscope, all of those in particular construed as an inverted microscope.

The present invention provides assistance for visual observation based counting of particles or crystals. In an image of a specimen of an unidentified sample, particles are counted from an image obtained as a result of binarization using a method such as the discriminant analysis method, for example, and crystals are counted from two images using the difference between the images according with different imaging conditions. As an example, in the particle counting, such as a two-step noise removal is performed, and also, for the crystals, alignment and aspect ratio calculation are performed. In particular, the present invention can provide assistance for the dispersion staining method in which asbestos crystals are visually searched for using a phase-contrast microscope.

The invention discloses a mould image recognizing method and a device thereof. The method comprises the following steps that A, a shot image of a sample is obtained, the edges of the sample image are recognized, and threshold binaryzation is automatically conducted on the sample image; B, the areas of all outlines in the binarized sample image are calculated, a calculating result is compared with a preset outline area value, and the image is processed according to a comparison result, so that outlines which meet the requirements are kept in the sample image; C, all the outlines in the processed sample image are traversed, gray level images of the outlines are cut out, an average gray level value of the gray level images is calculated, the calculated average gray level value is compared with a preset gray level threshold, and outlines which are larger than or equal to the gray level threshold in the sample image are kept. A phase contrast microscope is adopted for image shooting, and by means of a circular recognition method, mould in secreta of a vagina of a female can be recognized, so that the recognition rate of detection is high, and the detection efficiency is also high; full-automatic recognition is adopted, so that the detection cost is lowered.

A liquid surface in a culture vessel is irradiated with liquid-surface-measurement illumination light, and transmitted light that has passed through the liquid is detected by an imaging unit. A relative positional relationship between a focal plane of an image forming optical system and the culture vessel is changed, a detection signal for each position of the focal plane is obtained, and a liquid surface shape is estimated on the basis of the detection signal for each position of the focal plane. Then, on the basis of the estimated liquid surface shape, adjustment information for adjusting the optical characteristics of an adjustment optical system for adjusting refraction of light due to the liquid surface shape is acquired. After the optical characteristics of the adjustment optical system have been adjusted on the basis of the adjustment information, an image of a specimen is captured by irradiating the culture vessel with phase-contrast-measurement illumination light.

The present invention relates to an eyepiece base unit and a microscope with which a phase contrast observation function can be easily added to the microscope.The eyepiece base unit 12 is removably attached to a main unit of the microscope, and includes, in a state of being attached to the microscope, a pupil conjugate plane, which is a plane conjugate with an image side focal plane of an objective lens in an observation optical system of the microscope. By rotating a turret 32 around a central axis 67, phase plates 64a and 64c installed in the turret 32 can be inserted into the pupil conjugate plane. The present invention can be applied to a phase contrast microscope, for example.

The invention discloses a wheat root tip chromosome production method. The method includes the specific steps that after seeds germinate, rootlets are taken, preprocessed with 8-hydroxyquinoline and then subjected to fixation with Kano fixing fluid, low-osmolarity preprocessing with distilled water, enzymolysis with cellulase and pectinase and low-osmolarity postprocessing with distilled water, the processed root tip growth points are subjected to chromosome production through a smearing method, and finally microscopic examination is conducted under a phase contrast microscope. The chromosome production method is convenient and easy to operate, chromosome dispersity is good, the background is relatively shallow, counting is easy, and a splitting cell is not prone to losing.

The invention discloses a method and a system for detecting and screening quasi-circular cell regions to solve the problem that the quasi-circular cell regions cannot be effectively detected and screened in the prior art. The method comprises: S1: defining grayscale hierarchical structure information with different importance in the phase contrast microscope cell image; S2: determining cell main information of the current frame by means of manual marking or inter-frame correlation; S3: performing Hough circle detection for the original image and obtaining the position parameters and the radiusparameters of each Hough circle; and S4: screening the results of the Hough circle detection by comparing the relationship between the main information of each cell and each Hough circle region. Themethod and the system are based on the grayscale hierarchical structure information. By comparing the relationship between the main information of each cell and each Hough circle region, the results of the Hough circle detection can be effectively distinguished and screened, thereby facilitating the determination of cell morphology and its changes.

The invention relates to a method for detecting content and / or categories of medicinal montmorillonite impurities. The method includes: taking a montmorillonite sample, using water separation method or heavy liquid separation method to completely separate impurities from the montmorillonite, drying, weighing and calculating impurity content; subjecting the impurities to X-ray powder diffraction diagram, and judging categories of minerals in the sample according to the characteristic peak of X-ray diffraction; and using one of the polarizing microscope method, the phase difference microscope method, the scanning electron microscope method or the transmission electron microscope to confirm crystal habits of the impurities. Using the method which is simple can accurately detect content and categories of the impurities in the montmorillonite and solve the technical problem that existing methods for detecting impurity content of montmorillonite are inaccurate.

The invention provides a sperm-oocyte interaction vitro detection method, comprising: a. extracting sperms by a swim-up method or density gradient method; b. mixing the sperms and oocyte extracted from the step a according a certain proportion in the culture medium and setting a blank control of the oocyte in the culture medium; c. removing the sperms which are bonded with the surface of the oocyte in the step b and detecting the number of sperms bonded with the transparent area of the oocyte under an inverted phase contrast microscope; d. separating the oocyte from the sperms bonded with the transparent area of the oocyte and respectively collecting the separated sperms and the oocyte; e. coating the sperms in the blank control of the oocyte and sperms separated by the step b on the glass slide, drying and fixing the sperms using the fixation fluid, cleaning the fixed sperms and adding fluorescein-agglutinin label, re-cleaning the sperms after reaction, detecting the sperm acrosome reaction rate under a fluorescence microscope; f. detecting the number of sperms penetrating into the oocyte separated by the step d under the inverted phase contrast microscope. The detection method reduces the consumption of the label resource and the detection cost and time and increases the detection efficiency.

The invention belongs to portable temperature control cone board shearing pool; it is a device for research the crystallization and separation behavior of object combining with micropolariscope, phase difference microscope, fluorescent microscope, laser scattering and X ray scattering technologies under even shearing field, the research with the device to polymer system can provide reliable gist and theory instructions for designing of polymer material.

A single-axis illumination system for a multiple-axis imaging system, particularly an array microscope. A single-axis illumination system is used to trans-illuminate an object viewed with an array of imaging elements having multiple respective axes. The numerical apertures of the imaging elements are preferably matched to the numerical aperture of the illumination system. For Kohler illumination, the light source is placed effectively at the front focal plane of the illumination system. For critical illumination, the light source is effectively imaged onto the object plane of the imaging system. For dark field illumination, an annular light source is effectively provided. For phase contrast microscopy, an annular phase mask is placed effectively at the back focal plane of the objective lens of the imaging system and a corresponding annular amplitude mask is provided effectively at the light source. For Hoffman modulation contrast microscopy, an amplitude mask is placed effectively at the back focal plane of the objective lens of the imaging system and a slit is provided at a source of light of the illumination system. Structured illumination and interferometry, and a secondary source, may also be used with trans-illumination methods and apparatus according to the present invention.

An optical microscope is provided with an adjustable optical phase ring. The adjustable ring provides a way to compensate for distortion in the visible phase ring before the light reaches the sample. In an inverted microscope, when observing transparent cells under a liquid, the visible light phase ring is distorted. By the use of a Liquid Crystal Display (LCD) in place of a fixed ring, the projected ring is adjusted to realign the light and produce phase. In a typical micro plate, the meniscus formed produces a lens effect that is realigned by providing changes in the position and pattern, to allow phase imaging over a wider portion of the well. The realignment of the ring can be manual or automated and can be dynamically adjusted based upon an observed image of the sample.

A system and method of generating and acquiring phase contrast microscope images while minimizing interference with the intensity and optical quality of other microscopy modalities employing polarization and attenuation strategies for phase microscopy applications. A plane polarizing objective phase ring may be used in conjunction with a phase microscopy apparatus. Attenuated light may be controlled such that transparency may be selectively provided with respect to light in a predetermined plane. Illumination outside of the predetermined plane may be selected for phase microscopy applications. Accordingly, a polarizing objective phase ring effective for enabling polarized phase microscopy may reduce interfere with normal usage of the microscope for other applications such as, for example, fluorescence microscopy.