EP3803495A1 - Analyseur destiné à l'analyse tridimensionnelle d'un échantillon médical au moyen d'une caméra à champ lumineux - Google Patents

Analyseur destiné à l'analyse tridimensionnelle d'un échantillon médical au moyen d'une caméra à champ lumineux

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Publication number
EP3803495A1
EP3803495A1 EP19728133.0A EP19728133A EP3803495A1 EP 3803495 A1 EP3803495 A1 EP 3803495A1 EP 19728133 A EP19728133 A EP 19728133A EP 3803495 A1 EP3803495 A1 EP 3803495A1
Authority
EP
European Patent Office
Prior art keywords
cell
microscope
camera
sample
analyzer
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
EP19728133.0A
Other languages
German (de)
English (en)
Inventor
Thomas Engel
Gaby MARQUARDT
Gabriele Hörnig
Lukas RICHTER
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Siemens Healthcare Diagnostics Inc
Original Assignee
Siemens Healthcare Diagnostics Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Siemens Healthcare Diagnostics Inc filed Critical Siemens Healthcare Diagnostics Inc
Publication of EP3803495A1 publication Critical patent/EP3803495A1/fr
Pending legal-status Critical Current

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Classifications

    • G—PHYSICS
    • G06—COMPUTING; CALCULATING OR COUNTING
    • G06T—IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
    • G06T7/00—Image analysis
    • G06T7/50—Depth or shape recovery
    • G06T7/55—Depth or shape recovery from multiple images
    • G06T7/557—Depth or shape recovery from multiple images from light fields, e.g. from plenoptic cameras
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/36—Microscopes arranged for photographic purposes or projection purposes or digital imaging or video purposes including associated control and data processing arrangements
    • G02B21/365—Control or image processing arrangements for digital or video microscopes
    • G02B21/367—Control or image processing arrangements for digital or video microscopes providing an output produced by processing a plurality of individual source images, e.g. image tiling, montage, composite images, depth sectioning, image comparison
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/36—Microscopes arranged for photographic purposes or projection purposes or digital imaging or video purposes including associated control and data processing arrangements
    • G02B21/361—Optical details, e.g. image relay to the camera or image sensor
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B27/00—Optical systems or apparatus not provided for by any of the groups G02B1/00 - G02B26/00, G02B30/00
    • G02B27/0075—Optical systems or apparatus not provided for by any of the groups G02B1/00 - G02B26/00, G02B30/00 with means for altering, e.g. increasing, the depth of field or depth of focus
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B30/00—Optical systems or apparatus for producing three-dimensional [3D] effects, e.g. stereoscopic images
    • G02B30/10—Optical systems or apparatus for producing three-dimensional [3D] effects, e.g. stereoscopic images using integral imaging methods
    • G—PHYSICS
    • G06—COMPUTING; CALCULATING OR COUNTING
    • G06T—IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
    • G06T11/00—2D [Two Dimensional] image generation
    • G06T11/001—Texturing; Colouring; Generation of texture or colour
    • G—PHYSICS
    • G06—COMPUTING; CALCULATING OR COUNTING
    • G06V—IMAGE OR VIDEO RECOGNITION OR UNDERSTANDING
    • G06V20/00—Scenes; Scene-specific elements
    • G06V20/60—Type of objects
    • G06V20/69—Microscopic objects, e.g. biological cells or cellular parts
    • G—PHYSICS
    • G06—COMPUTING; CALCULATING OR COUNTING
    • G06V—IMAGE OR VIDEO RECOGNITION OR UNDERSTANDING
    • G06V20/00—Scenes; Scene-specific elements
    • G06V20/60—Type of objects
    • G06V20/69—Microscopic objects, e.g. biological cells or cellular parts
    • G06V20/698—Matching; Classification
    • H—ELECTRICITY
    • H04—ELECTRIC COMMUNICATION TECHNIQUE
    • H04N—PICTORIAL COMMUNICATION, e.g. TELEVISION
    • H04N23/00—Cameras or camera modules comprising electronic image sensors; Control thereof
    • H04N23/56—Cameras or camera modules comprising electronic image sensors; Control thereof provided with illuminating means
    • H—ELECTRICITY
    • H04—ELECTRIC COMMUNICATION TECHNIQUE
    • H04N—PICTORIAL COMMUNICATION, e.g. TELEVISION
    • H04N23/00—Cameras or camera modules comprising electronic image sensors; Control thereof
    • H04N23/60—Control of cameras or camera modules
    • H04N23/67—Focus control based on electronic image sensor signals
    • H—ELECTRICITY
    • H04—ELECTRIC COMMUNICATION TECHNIQUE
    • H04N—PICTORIAL COMMUNICATION, e.g. TELEVISION
    • H04N23/00—Cameras or camera modules comprising electronic image sensors; Control thereof
    • H04N23/90—Arrangement of cameras or camera modules, e.g. multiple cameras in TV studios or sports stadiums
    • H—ELECTRICITY
    • H04—ELECTRIC COMMUNICATION TECHNIQUE
    • H04N—PICTORIAL COMMUNICATION, e.g. TELEVISION
    • H04N23/00—Cameras or camera modules comprising electronic image sensors; Control thereof
    • H04N23/95—Computational photography systems, e.g. light-field imaging systems
    • H04N23/957—Light-field or plenoptic cameras or camera modules
    • G—PHYSICS
    • G06—COMPUTING; CALCULATING OR COUNTING
    • G06T—IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
    • G06T2207/00—Indexing scheme for image analysis or image enhancement
    • G06T2207/10—Image acquisition modality
    • G06T2207/10052—Images from lightfield camera
    • G—PHYSICS
    • G06—COMPUTING; CALCULATING OR COUNTING
    • G06T—IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
    • G06T2207/00—Indexing scheme for image analysis or image enhancement
    • G06T2207/10—Image acquisition modality
    • G06T2207/10056—Microscopic image
    • G—PHYSICS
    • G06—COMPUTING; CALCULATING OR COUNTING
    • G06T—IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
    • G06T2207/00—Indexing scheme for image analysis or image enhancement
    • G06T2207/30—Subject of image; Context of image processing
    • G06T2207/30004—Biomedical image processing
    • G06T2207/30024—Cell structures in vitro; Tissue sections in vitro
    • G—PHYSICS
    • G06—COMPUTING; CALCULATING OR COUNTING
    • G06V—IMAGE OR VIDEO RECOGNITION OR UNDERSTANDING
    • G06V2201/00—Indexing scheme relating to image or video recognition or understanding
    • G06V2201/03—Recognition of patterns in medical or anatomical images
    • H—ELECTRICITY
    • H04—ELECTRIC COMMUNICATION TECHNIQUE
    • H04N—PICTORIAL COMMUNICATION, e.g. TELEVISION
    • H04N23/00—Cameras or camera modules comprising electronic image sensors; Control thereof
    • H04N23/10—Cameras or camera modules comprising electronic image sensors; Control thereof for generating image signals from different wavelengths

Definitions

  • the invention is in the field of automatic analyzers and relates to a hematology analyzer for analyzing cells in a sample using a microscope apparatus having a light field camera.
  • automated cell counters For automated analysis of cells, so-called “automated cell counters” are used with increasing success. Examples include the Advia 2120, Sysmex XE-2100, CellaVision DM96 and CellaVision DM1200 device. These automated devices, apart from their high throughput, provide some advantages, such as high objectivity (no variability depending on the observer), elimination of statistical variations, which are usually associated with manual counting (counting high cell counts), as well as the determination of numerous parameters that would be unavailable on a manual count and, as mentioned, more efficient and cost-effective handling. Some of these devices can handle 120 to 150 patient samples per hour.
  • automatic Einzelzellzäh ment usually based either on an impedance (resistance) measurement or on an optical system (scattered light or absorption measurement). Further, imaging systems are established, e.g. Automatically make and evaluate cells of a blood smear.
  • cell counting and sizing are based on detection and measurement of changes in electrical conductivity (Resistance) caused by a particle moving through a small opening.
  • Particles such as blood cells, are themselves non-conductive, but are suspended in an electrically conductive diluent.
  • the impedance (resistance) of the electrical path between the two electrodes located on either side of the aperture temporarily increases.
  • the optical method involves passing a laser light beam or an LED light beam through a dilute blood sample that is in continuous flow from the laser beam or the LED
  • Light beam is detected.
  • the corresponding light beam may be e.g. be conducted to the flow cell by means of an optical waveguide.
  • Each cell that passes through the detection zone of the flow cell scatters the focused light.
  • the scattered light is then detected by a photo detector and converted into an electrical impulse.
  • the number of pulses generated here is directly proportional to the number of cells passing through the detection zone in a particular period of time.
  • the light scattering of an individual cell passing through the detection zone is measured at different angles.
  • the Streuver hold the respective cell for the optical radiation he summarizes that allows conclusions about eg the cell structure, shape and reflectivity.
  • This scattering behavior can be used to differentiate different types of blood cells and to use the derived parameters to diagnose deviations of the blood cells of this sample from a standard obtained, for example, from a variety of normal classified reference samples.
  • today's analyzers use microscopes with a high numerical aperture and with immersion between the slide and the objective to achieve a high resolution.
  • the usual cameras have pixel numbers of max. 0.3 to 1 million pixels. As a result, the field of view in the object is only a few 100 gm.
  • the surface of the area of the smear to be analyzed must then be scanned with a displacement unit in a meander scan procedure.
  • a first area scan with low magnification eg 10 times with a correspondingly 10 times larger field of view is performed and after a first image analysis to find the cells to be measured then only the regions of interest (Rol) with the Cells with the higher magnification subsequently targeted targeted.
  • the blood smears are stained in an upstream step. Several dyeing protocols have been established worldwide, some of which differ globally from region to region. Thus, the comparability of the analysis of blood smears is regionally limited, since only images of cells which have been stained according to the same protocol can be compared well.
  • An object of the invention is thus to provide an automatic analyzer for analyzing cells in a sample and a method for determining a two-dimensional or three-dimensional information of a cell, wherein, for example, chemical pretreatment of the cell should be avoided as far as possible before image acquisition and so that the analysis takes place around the cell in one to investigate the situation as originally and least or not at all changed.
  • the invention particularly comprises an analyzer for analyzing cells in a sample, the analyzer comprising an optical microscope for imaging a light field in an object area and / or an object plane for imaging cells of the sample, the microscope comprising a
  • the digital recording device preferably comprises a detection device for the light beams and allows a conversion of the detection signals of the detection device into digital data.
  • An analyzer according to the invention has the advantage that the hitherto necessary staining of the samples for the subsequent microscopy can be completely or partially dispensed with.
  • the analyzer according to the invention enables image data of the highest quality with correspondingly high information content, which can be used, for example, for automatic evaluation and classification.
  • the recorded image data have such high quality, so that a follow-up examination of the sample by renewed microscopic examination is no longer necessary.
  • a complete digitizing station of the cell is provided with a high resolution for lateral structures and also over the entire depth extent of the respective cell. In particular, a coloring and / or marking of the blood cells for the image acquisition is not necessary.
  • the analyzer according to the invention is preferably an automatic analyzer, particularly preferably a semi-automatic or fully automatic analyzer.
  • the microscope preferably comprises the light field camera.
  • the digital recording device Before given to the light field camera includes the digital recording device, which is designed to receive the imaged in the microscope light field.
  • the digital recording device comprises a charge-coupled device (CCD) chip or a plurality of CCD chips.
  • the digital recording device is based on a complementary metal-oxide semiconductor (CMOS) technique and / or comprises a CMOS chip.
  • CMOS complementary metal-oxide semiconductor
  • the sample is transported within the automatic analyzer fully automatically, e.g. by ent speaking flow systems and / or experience of sample carriers within the analyzer, preferably by means of appropriately controlled actuators or robot systems.
  • the light field camera comprises a microlens array with lenses of different focal lengths, the microlin senarray an intermediate image of the imaged in the microscope
  • the microlens array is at least one focal distance from the intermediate image removed and there is a real Abbil tion.
  • the picture information becomes then projected back through the optics until the rays of corresponding pixels from different sub-images meet. In the present case, therefore, no images of the images are taken but small object images.
  • the invention is based on an optical microscope, which is equipped with game contrast with devices for differential interference contrast.
  • Such microscopes can be adapted to the requirements of hematology in reference to the splitting. This adaptation takes place essentially by a specific choice of the beam offset on the beam away through the measurement object.
  • the measurement object is, for example, a crossed out blood sample.
  • the beam offset is determined by the thickness of the DIC prisms in the illumination module and in the analyzer module.
  • the optical thickness of the two prisms and the direction of the beam offset must be coordinated with each other, so that the analy satormodul the beam offset from the lighting module can completely reverse and thus compensate. Due to different magnifications, the physical thicknesses of the prisms can deviate from each other. For microscopes, there may be several pairs of DIC prisms for one lens, allowing for different splitting.
  • a light field camera also called a plenical camera, is provided on the microscope, which records the light field formed in the microscope from the object plane.
  • the light field camera has an effective aperture number (working f #) in the range of 10 to 30, particularly preferably it is 26.
  • working f # effective aperture number
  • a suitable magnification is selected for this purpose, the sorelement the size relationship of the real Sen, that in the range of typically 1 to 10 pm and the scanning of the object, which is in the range of 0.05 to 0.5 gm, which is preferably because of the high resolution requirements in the measurement of the range of 0.05 gm to 0.15 pm is taken into account.
  • the light field camera is preferably a light field camera with the designation Raytrix R12 Micro Series from Raytrix GmbH, Kiel.
  • the image of the object can be reconstructed for different depths, which corresponds to different focus settings of the microscope. This purely digital refocusing can be done retrospectively on the recorded data sets.
  • the analyzer according to the invention has the advantage that, in addition to the color information RGB, depth information D (depth) which is available in a downstream computer-based Evaluation as a further feature analogous to a color channel ver can be used.
  • the cells may e.g. segmented more easily, so recognized for further analysis and cut out of the picture.
  • contrasting color transitions then arise in the height profile at the edges of the cell transitions that are well and accurately detectable.
  • the captured image e.g. the blood smear or the cells
  • This can, on the one hand, be used to segment the cells and identify them for classification
  • volume data or 3D point clouds can also be used. This allows the segmentation and Klassifika tion done with higher accuracy. If the images, eg of the cells, have been taken with the light field camera, it is possible subsequently to focus the images purely digitally through, ie offline. This would allow for unclear classifications or evaluations of cells by the computer, the doctor the ability to look at the cells directly in the digital image and not just the slide again under a microscope to le gene, to search the cell and then on the variation a more accurate rating and classification.
  • the doctor can now position the cell to be examined only with an approximate location in the microscope and then has to search the cell in the final microscope. This is time consuming and must be successful in the laboratory.
  • the digital image of the light field camera according to the present invention is present, the physician can directly focus the cell or structure in question in the digital image.
  • This evaluation and classification is then also in the sense of telemedicine in an existing data connection of the doctor to the image or the image database possible so that he no longer has to come to the slide in the laboratory.
  • a consultation with a colleague can be made, who receives the image for analysis or, for example, makes it available via electronic networks, which can be done virtually in real time with digital data transmission.
  • the respective physician only needs a corresponding analysis software on his digital terminal or in the sense of a cloud solution, both the image of the light field camera can be stored in the cloud and the evaluation, for example via a web - Browser Interface suc in a cloud-based application.
  • the magnification is determined from the requirement of the optical resolution for the scanning of the measurement object, wherein with the choice of magnification in general, the Benö required effective f-number or the numerical aperture is a related party.
  • the lateral resolution is preferably 100 nm in the object level.
  • the magnification then results together with the lateral dimension of a sensor element
  • V lateral dimension sensor element / 100 nm.
  • This magnification is e.g. particularly advantageous for the formation of blood cells.
  • nm is the typical resolution limit of light microscopy in the case of water, oil or glycerin immersion.
  • a known pixel size of the camera e.g. 4.5 gm would be the required magnification 45x.
  • the f-number of the camera is preferably 2.4, 2.8, 5.6, 7.0 or 26.0. More preferably, the f-number of the camera is f # 26 and the magnification is 63 times.
  • the analyzer is preferably a hematology analyzer, particularly preferably an automatic hematology analyzer.
  • the medical sample comprises a cell and / or a medical preparation.
  • the medical preparation is preferably a tissue section, sediments of body secretions and / or body fluids and / or microcrystals.
  • the sample is a blood sample and / or the cell is a blood cell.
  • the sample may preferably be any type of human, animal or plant cell. This has the advantage that various types of probes, including various cell types, can be examined and characterized.
  • the microscope is preferably an amplitude contrast, and / or a phase contrast, and / or a differential interference contrast (DIC) microscope.
  • DIC differential interference contrast
  • the microscope is a differential digital holographic microscope.
  • the extended contrasting methods make it possible to visualize phase differences of the light paths through the sample, in this case, for example, the cells.
  • Different phase values in particular during phase contrast then, for example, with different colors be displayed and measured with a color camera who the. This makes it possible to dispense with a dyeing of the samples and still depict the cells well contrasted.
  • Previous systems for automated cell classification use exclusively the amplitude contrast, which results from the staining of the cells and the different paths of the light in the cell or alternatively the surrounding medium.
  • the combination according to the invention of a microscope and a light field camera gives additional 3D information about the cell, which can be used for a classification of the cell.
  • the classification may e.g. performed by healthcare professionals and / or computer-based systems.
  • amplitude contrast conventional image information and additional 3D information are present in the cell.
  • phase contrast a phase image and additional 3D information of the cell is obtained.
  • DIC a DIC image is obtained on differential phase images as well as additional 3D information of the cell.
  • the resulting 3D information of the cell may advantageously be e.g. be represented differently as follows.
  • the 3D information of the cell is displayed as a RGB image.
  • RGB image as so-called total focus image over the entire TiefenJrfebe empire of the light field camera is sharply displayed. Due to the increased depth of field compared to conventional 2D cameras, more depth information is captured.
  • a blood cell has - depending on the type and orientation - a thickness of about 1 to 2 gm, to about 20 gm.
  • the depth of field increases by at least the factor four on the plenoptic effect from the light field camera.
  • cells in particular, for example, red blood cells (RGB) are fully focused.
  • larger cells such as white blood cells (WBC), are imaged sharply to a considerably larger portion of the cell volume.
  • the 3D in formation of the cell is displayed as RGB D information.
  • Each pixel contains a depth information called D.
  • D is e.g. the thickness of the blood cell. This information complements the color information and is also referred to as a 3D point cloud.
  • the 3D in formation of the cell as volumetric 3D information Darge is. Analogous to an image of a computer tomograph (CT) ent spatial information thus in the form of voxels. Since the cells, e.g. Blood cells for which radiation is at least partially transparent, so different Stel len the cell generate stray radiation, which is recorded by the camera and assigned to different depths.
  • CT computer tomograph
  • an image stack can be calculated from the data set of the light field camera who the, which is calculated on different focal planes and thus contains the volumetric information.
  • the so-called virtual depth is the distance measure- ment.
  • the focal planes can also be calculated with other distance values, which are selected equidistantly, for example, and one level with the max. Cross-section of the cell falls together.
  • the image of the light field camera based only on the intensity modulations generated by the phase effect, since the amplitude or intensity modulation are filtered out of the object itself in the phase ring. This image is as colorfast as possible to the image with pure amplitude contrast.
  • the image of the light field camera is based on an image of the cells, which contains information about the different paths of the light as it passes through the cell and thus the phase change of the light as a color-coded information via the differential interference contrast.
  • This has the advantage that the color representation of the cell superimposes a fine structure, which is very helpful for the evaluation algorithms of the light field camera for a high lateral resolution in the calculation of the depth information.
  • the spectrum of illumination includes the visible range and / or near IR wavelengths.
  • a color balance of the light source to a camera such as the light field camera.
  • an adjustment of the exposure time and the gain for the 3-chip RGB color camera for the 2D images preferably takes place.
  • the light source comprises a 4 LED light source with RGB and W and a common brightness control.
  • a further camera for recording an image in an object plane in the object area is provided in the analyzer on the microscope, wherein the further camera has a lateral resolution which is equal to or preferably higher than the lateral resolution of the light field camera, wherein the resolution of the further camera is twice, preferably three times, particularly preferably four times the resolution of the light field camera.
  • the further camera preferably has a larger field of view, also referred to as field of view, as the light field camera. This allows a better and faster Probenabde ckung.
  • the focal plane of the further camera, before given to a 2D camera is coupled to an excellent plane of the light field camera. This can be measured, for example, in vir tual depth. So it can be advantageously enough that both cameras deliver good pictures at the same time.
  • the further camera is a color camera, preferably a color camera
  • Multi-chip color camera particularly preferably a 3-chip Farbka mera.
  • the high-resolution color camera such as a high-resolution RBG camera
  • a high resolution is guaranteed.
  • This is of particular advantage since, in principle, a factor of two is lost laterally in the light field camera, and a factor of four in terms of resolution is lost in order to obtain information for calculating the depth resolution.
  • This loss of resolution can be fully compensated before geous enough.
  • the use of a multi-chip color camera, such as a 3-chip color camera has the advantage that then a better color measurement and in particular pixel-by-pixel color determination without interpolation with more dynamics is possible, which brings more before parts for the classification of cells.
  • an adjustment of color channels takes place for an optimized S / N ratio.
  • the compensation is preferably carried out separately for each staining protocol.
  • the 3-chip color camera is a 3-chip CMOS camera with 3.2 megapixels per chip.
  • a 3-chip color camera is compared with e.g. a 1-chip color camera preferred because a 1-chip color camera usually use a Bayer pattern. Since, in the analysis of e.g. Blood cells in a hematology analyzer e.g. Particularly good color resolution and color fastness and good lateral resolution of the structure are the preferred features of the 3-chip camera.
  • the high resolution color camera provides e.g. a high resolution RGB image with typically better, i. more accurate color and less noise by separately optimizing the exposure for each of the color channels. In the case of the 3-chip color camera, the color values are determined directly and without interpolation for each pixel separately.
  • the color values of a color camera are subsequently transferred from the RBG representation to, for example, an HSV representation for color value (Hue), saturation (Saturation) and brightness (Value), then, for example, red blood cells, where the red Dye is homogeneously distributed in the cell, also well above the color value as a further or complementary method to segment.
  • This approach works well for all cells and structures that have the same color value (Hue) or are colored with the same color.
  • This segmentation can advantageously with the over the Depth values (D) are combined in order to have as precise a criterion as possible to excise the cell.
  • the images of the two cameras are advantageously registered via scaling and / or pixel interpolation to each other, if the effective pixel sizes in the images do not match or are commensurate with integer factor of e.g. 1, 2, 3, etc.
  • lateral displacements, distortions, tilting, distortion and / or distortion in the images and / or a defocus are in principle advantageously to be corrected. If the images are then registered to the required extent, new image data is calculated which combines the higher latent color resolution with the depth information from the image of the light field camera.
  • the image of Farbka mera can be set to the assigned focus position in the evaluated Tie fensent the light field camera and from there via the known from the recording of the light field camera Propa gation of the light field and the color representations and Lateral len resolutions transmitted to adjacent focal planes who the.
  • the transmission can take place, for example, laterally via interpolation, for the colors e.g. via correspondence tables.
  • a more elaborate transmission is also possible via a true propagation calculation based on the data measured by the light field camera together with e.g. a phase retrieval possible.
  • evaluated image planes of the light field camera serve as support points and the additional additional points are supported by the neighboring points.
  • this more accurate solution which is typically very computationally intensive, is used for fine diagnostics, eg on conspicuous image areas, cells with unclear findings and / or pathological cells, where less or no real-time capability is required.
  • the fine diagnostics can then also be done, for example, on the complete image or even only on image sections, for example, on individual segmented cells possibly extended by certain margins around there.
  • the two cameras can be used and operated simultaneously on the microscope. In living or moving cells or samples or examinations using microfluidic cells, it may be advantageous if the two cameras also record the images synchronously in time.
  • microscope tubes which mechanically enable the parallel operation of two cameras on a microscope.
  • beam splitting e.g. the splitting ratio, the splitting method such as spectral separation, polar separation, etc., which can be chosen to be advantageous.
  • the division of the light field on the imaging side is already taken into account in the orientation of the polarization before the splitting in the illumination module in such a way that after the DIC prism and DIC analyzer the two polarization components satisfy the desired intensity ratio, e.g. 1: 1 for maximum contrast. In this way, light losses can be minimized in order to optimize the efficiency of the overall setup and to minimize the required exposure times for a quick measurement.
  • the separation of the beam path takes place in the case of the cameras between the imaging lens and the DIC analyzer with a polarization-neutral beam splitter.
  • a polarizer is still to be arranged so that it blocks the one polarization direction of the DIC beam path as completely as possible and let the other pass with the least possible attenuation to the camera.
  • This polarizer can either work in transmission or in reflection or lead to a spatial separation of the differently polarized beams. This offers the advantage that the structure is less Comp complex in the design and no special polarization-dependent adjustment needed, however, go principle, about 25% of the amount of light lost.
  • the images of the light field camera and the color camera are mutually aligned and scaled so that, for example, in a topography image, the 3D or Height information of the light field camera can be used and at the same time the color information can be used and used by the color camera.
  • the recording takes place by means of the light field camera and the color camera at the same time.
  • This can e.g. be achieved by ent speaking triggering.
  • the triggering can be implemented via hardware or via software.
  • the triggering can also take place from a selected first camera to the second camera, which is also referred to as master-slave coupling. Functionally it must be ensured that the pictures are taken with a given temporal reference, ideally at the same time.
  • the trigger signals may themselves have a time interval.
  • the same time exposure is of particular advantage for the uptake of moving cells, e.g. in a flow cell, and / or living cells or measurement objects.
  • only one polarization tion direction is used for the color image and the separation of the beam paths for color camera and light field camera is still done in front of the DIC analyzer or in front of an objective-side DIC prism.
  • the division of the light field can be advantageously considered in the direction from the polarization prior to the splitting in the lighting module so that after the DIC prism and DIC analyzer, the two polarisati onsanteile the desired intensity ratio, such as 1: 1 for maximum contrast, have. This is especially advantageous if stained cells are to be imaged and analyzed.
  • the color camera for high-resolution DIC and the plenary optics for an extended depth measurement range for unambiguous unwrapping the color ranges for measuring thicker cells or Zellclus tern is used.
  • a high-resolution relative thickness measurement therefore, it follows by means of DIC in that the differential Kontrastin formations, for example. be integrated and a more spacious assignment is done by plenipotentics.
  • the amplitude contrast barely contains no information and the color image is advantageously used only for phase or DIC contrast, but then has a high lateral resolution with a true color separation per pixel.
  • the light field camera and the high-resolution camera which advantageously comprises a color camera, are aligned relative to one another with respect to the axial directions of the sensor array and advantageously also the subpixel accurate shift along the axis directions of the array and / or a directional scaling factor for e.g. the x and / or y axis.
  • the position of the focus of the high-resolution camera on the average measuring range of Light field camera set is advantageously improved.
  • f # the achievable depth resolution of the light field camera increases.
  • a f-number of 7 can be significantly more advantageous than a Blen denress 26.
  • the object with the full imaging aperture is already illuminated on the microscope in order to illuminate on the image side the full aperture of the working f # so that the light field camera works well and can provide a high lateral scan.
  • the incoherent illumination Sigma is greater than 0.8, preferably greater than 0.9, be particularly preferably 1.0, where sigma is given as the Quoti ent from the numerical aperture of the illumination at the microscope the sample and the numerical aperture of the objective, and wherein the microscope is preferably a differential interference contrast (DIC) microscope.
  • DIC differential interference contrast
  • the analyzer comprises a Probenzu arrangementseinrich device for slides by means of the analyzer samples can be supplied to a slide.
  • the analyzer comprises a flow cell for the supply of the sample, wherein preferably the object level of the Mik roskops is located in the flow cell.
  • the flow cell is a microfluidic flow cell.
  • the usable layer is typically several microns thick in a flow cell, sometimes even some 10 gm thick, it is difficult for the cells to precisely position in the object plane of the microscope used for the examination. In this regard, there is a conflicting requirement tion of the desire for high resolution on the one hand and the not so precise position of the cell in the direction of the optical axis on the other.
  • the depth measurement of the light field camera can preferably be used here for optimizing the activation parameters of the microfluidics, in order, for example, to adjust the focus appropriately so that the 2D high-resolution images are also sharp.
  • An exposure and 3D images is also an added value without interferometry, as in DHM, which is complex and complex and currently. Interference by effects with temporally and / or spatially coherent radiation has.
  • the depth measuring range ie the area where sharp images are recorded
  • the lateral resolution of the light field camera decreases by a factor of 2.
  • magnification and the aperture can be selected largely independently of one another in the selection of the optics used, however, this effect can be well compensated. If one chooses e.g. a larger magnification and a higher aperture, so the effect can be correspondingly positive ver.
  • NA 0.7
  • d 0.36 gm
  • depth of field d 1.05 pm.
  • the depth working range can thus be greatly increased, which then makes it possible to use a flow cell accordingly.
  • a more complete or complete detection of the expansion of a cell via the height extension of the flow through the flow cell is possible. This allows in particular precise investigations on typically unstained cells. For certain studies, however, it may also be advantageous or necessary for the cells to be stained in the medium of microfluidics.
  • a further advantage is that the parameters of the flow cell with regard to focusing and / or position of the cell can lie in a further parameter range and e.g. can be dispensed with a corresponding special optimization.
  • the field of view of the microscope includes the full width of the flow cell through which cells can flow.
  • This area advantageously has a width of a few 1/10 mm to a few mm.
  • the analyzer comprises both means for viewing slides with cells and / or for viewing cells in the flow cell.
  • the dimensions of the flow cell on the dimensions of the object are carriers, in particular their thickness and thus the optical effects We matched.
  • Flow cell advantageously also have optically the same effect, e.g. with regard to optical path length, dispersion and / or refractive index, so that the same optics can be used and the same best possible imaging quality is maintained.
  • transfusion may be performed to obtain e.g. to adapt to manufacturing tolerances.
  • Typical cover glass thicknesses are in the range below 0.2 mm, typically 0.15 to 0.17 mm.
  • the lateral dimensions of slides are, for example, 76 mm by 26 mm or 75 mm by 25 mm according to DIN ISO 8037-1 at one
  • Thickness in the range of 1 mm to 1.5 mm. The dimensions of the flow cell then arise advantageously accordingly.
  • the speed of movement of the cells in the flow cell in the imaging region of the microscope is adapted to the exposure time of the image acquisition systems used.
  • the movement is typically smaller than U pixels (pxl), preferably smaller than 1/5 px, particularly preferably smaller than 1/10 pxl. This has the advantage that motion blurring effects can be reduced or completely avoided.
  • the analyzer comprises a sample feeding device for slides. This may be particularly advantageous when e.g. Tissue sections or other medicinal preparations should be examined.
  • Another object of the invention is a method for determining a two-dimensional or three-dimensional Information of a cell by means of an analyzer according to the invention, the method comprising the steps
  • step d) determination of a two-dimensional or three-dimensional and / or volumetric information of the cell from information recorded in step b) of the light beams and / or information of the light field imaged in step c).
  • steps b) and c) may also be combined into one step, the step then comprising
  • step b) the light beams emanating from the cell in the illuminated sample are preferably first focused and then drawn by means of the digital recording device.
  • a conversion of the photons into electrical charge with subsequent determination of a charge quantity and digitization of the charge quantity is preferably carried out.
  • a two-dimensional or three-dimensional information can preferably also be determined correspondingly from a medical preparation.
  • the sample is preferably supplied to the analyzer in step a) by means of a flow cell, the object plane of the microscope lying in the flow cell.
  • the flow cell comprises means making it possible, by appropriate control of the flow cell, to enrich the cells in an excellent plane in the flow cell.
  • the object plane is set to the excellent plane, so that the object plane and the plane drawn out advantageously coincide.
  • the sample is supplied in step a) by means of a sample feeding device for slides to the analyzer on a slide.
  • the method further comprises a step in which an image is provided in an object plane in the object region by means of a further camera for taking an image in the object plane in the object region, wherein the further camera has a lateral resolution which is higher than the lateral resolution of Light field camera is, wherein the resolution of the wei direct camera is preferably twice, particularly preferably four times the resolution of the light field camera and wherein be preferably the other camera has a larger field of view than the light field camera.
  • the further camera is a color camera, preferably a 3-chip color camera.
  • the cells and / or the medical preparation are preferably not stained.
  • the extended contrasting methods such as, for example, phase contrast and / or differential interference contrast, in particular for imaging cells, are particularly advantageous.
  • amplitude contrast may also be advantageous.
  • the method further comprises the step
  • step d) digital refocusing or focus variation by means of the determined in step d) two-dimensional or dreidi dimensional and / or volumetric information of the cell along the optical axis of the microscope, wherein preferably the digital refocusing is computer-aided and / or nume risch.
  • the corresponding object carrier with the questionable cells has been physically brought out of eg a magazine, a storage box or an archive out and then placed under a microscope and the image ever Weil focused on the respective cell. Based on relatively inaccurate location information with respect to the position of the respec gene cells on the slide was then tried to find each respective cells again. If the cells were found again, the diagnosis is made. To determine this, the doctor analyzes the optical image of the cells and usually focuses through the cell. Therefore, the previously known systems and methods for de waisted Nachbefundung are not or only very suitably appro net, as the doctors no longer taken once the cell image be able to focus and thus an important for the diagnosis depth information is not accessible.
  • the cell image can be recorded as a three-dimensional image in order to allow the doctor on the digitally stored image a freely adjustable and at any time, also subsequently selectable, focus on different focal planes.
  • the doctor no longer has to sit on a microscope himself, but the process can be performed spatially and temporally independent of the sample. Since the image is purely digital, occurs over the time since the image was taken no deterioration of the data, in contrast to the hitherto customary procedures in which the archived samples age over time and deteriorate in their state. For each follow-up, the sample would be returned to the microscope and immersed in immersion oil. After diagnosis, the sample is then cleaned again, which can also lead to damage and overall represents a certain time-consuming effort.
  • Another advantage is that when multiple cells are on an image, the focus can be adjusted and varied sequentially with respect to each one of the respective imaged cells.
  • the method further comprises the step
  • a further subject of the invention is a method for assigning a cell to a cell type, comprising a method according to the invention for determining a two-dimensional len and / or three-dimensional and / or volumetric Infor mation of the cell, wherein at a first location, the steps a) to d ), and wherein the information determined in step d) is transmitted digitally to a second location via a data and / or network connection, and wherein steps e) and f) are performed at the second location.
  • the invention also relates to a corresponding method for assigning a cell to a cell type, wherein, however, ever determining a two-dimensional or dreidimensio nal or volumetric information of the cell using any other suitable method for determining a saudi dimensional or three-dimensional and / or volumetric information Cell is done.
  • a worldwide learning system is etab lated. This makes it possible for larger data sets to be collected as quickly as possible, even for very rare disease pictures. These data sets can then be used advantageously also for automatic computer learning algorithms, so that ultimately also for the automated evaluation and / or assignment of the cells a broader and more secure basis is available.
  • the patient data are kept in a database.
  • This has the advantage that even later onset of disease can be analyzed earlier records on blood images and can be examined for abnormalities, such. in the case of hematology, to detect or recognize very early stages of development or signs of leukemia. If these data are learned by a system, this analysis can be automated on the images and run without additional effort, since the cell images are advantageously pre-evaluated by computers.
  • the presentation of the cells and / or samples preferably takes place by means of a 3D display device, preferably e.g. an autostereoscopic 3D display.
  • a 3D display device preferably e.g. an autostereoscopic 3D display. This allows the physician to give a novel 3D visual impression of the cells and / or samples to be examined. In particular, this advantage is also achieved in telemedicine according to the invention.
  • a compact data format for the image data from the light field camera is used.
  • who compresses the image data is used. This makes it possible to keep the required storage space as small as possible, which is only the Storage of many patient records allows, which can then be used as a ground truth, for example, for a computer-learning system in terms of automated medicine and / or as a valuable assistance system for doctors.
  • cloud solutions and / or server solutions for image storage and data of the diagnosis and advantageously also for the backup of extended data, which person, e.g. Doctor, when and on which system with which configuration or software version the cell in question has been diagnosed or also when and where the sample in question was taken from the patient and information about the transport to the laboratory.
  • this information may preferably also be provided via a link to another storage system.
  • the time of Befun training is stored accordingly.
  • Another object of the invention is a method for digital staining a cell, the method comprising a method according to the invention for determining a saudimensio dimensional or three-dimensional information of a cell by means of an analyzer according to the invention and additionally the following steps
  • the two-dimensional or three-dimensional and / or volumetric information of the cell is geometric information of the structure of the cell or medical preparation.
  • pixels under two-dimensional information, e.g. understood an image information that maps an object area in a planar image with the lateral coordinates in the X and Y direction, which describe the area and spans nen.
  • image information that maps an object area in a planar image with the lateral coordinates in the X and Y direction, which describe the area and spans nen.
  • the individual pixels of a two-dimensional image are also referred to as pixels.
  • three-dimensional image information e.g. ver stood that the image in addition to the areal Schminforma tion additionally at least for one pixel also contains a fen fen information in the axial direction of the Schmstrahlengan ges, so for example. in the Z direction, which is linearly independent of the X and Y directions.
  • An example of three-dimensional image information could be e.g. be a contour image that in the z-direction, the topography of the object as derelle.
  • volumetric information is used if the image also contains information over a large area in the X and Y directions, also in the Z direction for different Z values.
  • the X, Y and Z axes may each be divided with increments of equal size to produce small volume elements, also referred to as voxels.
  • Image information described via voxels is a form of volumetric information.
  • Volumetric image data are used eg in tomography. If the voxel information is present only for a few pixels or voxels, the information can also be represented in the form of so-called point clouds, where each of the points is, for example, over its X, Y and Z coordinates are represented.
  • the volumetric information is an extended representation for three-dimensional image information, which is particularly advantageous in the case of complex structured objects.
  • the two-dimensional or three-dimensional and / or volumetric information is information relating to intracellular structures such as e.g. Cell organelles and / or geometrical structures of a tissue section.
  • Information about optical path lengths generally does not readily conform to geometric information, and therefore optical path lengths are not geometric information in this regard.
  • the dyeing protocol is preferably the dyeing protocol May-Grünwald-Giemsa, Modified-Wright dyeing, Wright-Gimsa dyeing and / or Romanowsky dyeing.
  • the digital coloration of the cell it is preferable to use a color distribution according to one of the above-mentioned dyeing protocols.
  • images of the technical contrast of black and white and / or according to extended contrasting methods are preferably recolored by means of dyeing properties determined by computer learning.
  • 3D and / or volumetric information are preferably used, since this is advantageous over the use of a pure 2D color mapping.
  • the methods according to the invention for the digital staining of a cell have the advantage that the digital staining of the cells can be carried out subsequently in the digital image. This eliminates the additional steps of dyeing the south in the sample preparation. This leads to considerable time and cost savings. Furthermore, this avoids differences between different laboratories or regions.
  • Another advantage is that switching between different staining models is possible, e.g. to be more specific for certain pathologies.
  • the staining protocol is an artificial color model, e.g. emphasizes certain critical features in order to simplify the diagnosis for physicians, or in the first instance to enable the diagnosis of inexperienced doctors.
  • the artificial color model can be designed similarly to a false color representation for technical images, e.g. similar to thermal or IR images, which are displayed as a color image.
  • the staining models can be ge changed, which would not be possible with a classical chemical staining.
  • cells assigned to different classes are colored differently digitally.
  • all cells in the image that are currently not to be considered are hidden. Preferred is alternatively only a selected set of different cell groups are shown.
  • the 3D information of the cells possibly also in combination with the advanced contrasting methods such as phase contrast or DIC, used to replace the omitted Far binformation of attributable immediate staining by the 3D information.
  • the advanced contrasting methods such as phase contrast or DIC
  • the predetermined association between the two-dimensional or three-dimensional or volumetric information of the cell and staining of a corresponding cell and / or structure within the cell is determined by a staining protocol as follows in a learning phase.
  • the cell or the cells without staining in a first image is displayed. Subsequently, the cells are stained in accordance with the respective desired staining protocol. Then the stained cell or cells are stained in a second image. In the first and second images, the cell (or cells) is then recognized and segmented, and the unstained and stained cells are assigned accordingly.
  • the cells are grouped by cell type, e.g. red blood cells (RBC), white blood cells (WBC) (optionally inclusive 5 part diff.) and / or classified, e.g. by means of computer learning and / or neural networks.
  • cell type e.g. red blood cells (RBC), white blood cells (WBC) (optionally inclusive 5 part diff.)
  • WBC white blood cells
  • / or classified e.g. by means of computer learning and / or neural networks.
  • the color deviations can also be evaluated against an external reference standard, as described, for example, in US Pat. in DICOM for displays and display devices is described. Since the standard is not written for this application, evaluations may only be transferred and applied analogously.
  • the staining model is accordingly set, checked and stored for each staining protocol and each cell grouping individually or specifically.
  • the procedure is preferably as follows. First, the desired staining protocol is selected and the unstained cells are displayed in a first image. The cell or cells are segmented in the first image. Then an assignment of the cells to cell grouping and / or a determination of the cell type for the selection of a cell-specific staining model. In the next step, the cell or cells are stained. Subsequently, the display of a digitally colored image, e.g. a blood smear, on a dispenser.
  • a digitally colored image e.g. a blood smear
  • the same uncolored image in the pas send dyed each regionally common color. This allows the common evaluation of a sample by experts who are used to different colorations.
  • the digital staining of a cell according to the invention is preferably an image of a preferably uncolored cell taken digitally in an analyzer according to the invention. This image of the cell is then colored in a post-processing, as it is used to the doctor of the stained according to the different conventional staining protocols cells.
  • different staining protocols can also be selected according to the invention with the new digital staining for different cell types, because, for example, their specifics for the particular cell type are particularly suitable.
  • a doctor or hematologist or pathologist could create personal staining schemes where he assigns a specific type of staining to a cell type and completely breaks through and abolishes the previous restriction of just one staining for a slide.
  • generic digital staining protocols can advantageously also be created, which continue to use the technical color space for better recognizability of features and structures
  • Chemical dyeing works, in each dyeing step typically, essentially a dye which accumulates in un ferent amounts of suitable bindable cell structures.
  • the more dye attaches the more light is absorbed from the spectrum of the illumination light according to the absorption spectrum of the respective dye out biert.
  • the transmission image thus contains a higher color saturation (S) for the color value (s) of the dye (V).
  • S color saturation
  • V color value of the dye
  • the dye in addition to the pure color effect, the dye also has the desired side effect that the RNA still available in the cell clumps and only then can be imaged and contrasted in the micrososcope.
  • the undigested RNA is smaller in dimension than it would be resolvable with conventional optical microscopes.
  • the object of the invention is based on an optical micro-microscope, which is advantageously equipped for image acquisition with th th extender optical contrasting methods.
  • This Advanced contrasting methods may include, for example, phase contrast (PC), differential interference contrast (DIC), polarization contrast (POL), interferometry (preferred as a digital holographic microscope (DHM)), hyperspectral imaging (pure color contrast in an extended spectral range, eg UV, VIS, NIR, MIR and / or FIR or selected sub-areas thereof), and / or structured turator lighting (eg, with predetermined intensity distributions and / or phase distributions, preferably in the Pu pillenebene set).
  • PC phase contrast
  • DIC differential interference contrast
  • POL polarization contrast
  • interferometry preferred as a digital holographic microscope (DHM)
  • hyperspectral imaging pure color contrast in an extended spectral range, eg UV, VIS, NIR, MIR and / or FIR or selected sub-areas thereof
  • Different contrasting methods are preferably present in parallel on a microscope system, in particular if these are used, for example. mutually free of interaction usable. Otherwise, the different contrasting methods are preferably carried out in chronological succession. If the different contrasting methods are carried out in chronological succession, according to the invention a particel image velocimetry (PIF) is preferably provided for tracking the respective trajectories, since the cells move in a combination with a microfluidic system between the images Modalita obtained color information then again bring together for each cell correctly.
  • the PIV also includes the rotation of the cell and advantageously also the defocusing.
  • the samples are given e.g. a color contrast of interference, such as DIC or polarization, which then provides in the digital image for a good resolution of the structures in the cell and / o which allows a good contrasting for the cell.
  • a color contrast of interference such as DIC or polarization
  • a color information in addition to a 3D information is present.
  • the images of the light field camera for the 3D image can also be present as volumetric and / or tomographic data.
  • this 3D information also permits a subsequent purely digital focussing on the image taken. This is of particular advantage for the review of automatic classification by the physician and for the specific and reliable assessment of pathological or conspicuous cells in this context.
  • new dyeing modes are also provided in digital dyeing, where the color currently displayed depends only on the area of the cell which is e.g. is located below the focal plane.
  • the 3D depth perception of the doctor can be further improved.
  • only areas above the current focal plane can be used or the areas in a certain adjustable
  • a picture taken microscopically with one of the systems according to the invention is then postprocessed so that it looks as it would have looked like if it had been taken with a classical microscope of stained cells.
  • a certain number of unstained samples with a plurality of cells are taken up in the new system.
  • the cells can be detected and segmented manually or automatically in the image for comparison. Thereafter, the same samples are stained with the desired staining protocol and then resumed with a microscope the same cells.
  • the microscope has a good color fidelity and a color camera, particularly preferably a color camera optimized for the most color-fast image capture, which may also be equipped with special samples or Calibration procedures was calibrated, eg according to the DICOM standard in medicine.
  • Typical color cameras have a Bayer pattern in which 50 percent of the pixels are predominantly green, 25 percent of the pixels are predominantly blue, and the remaining 25 percent of the pixels are predominantly red.
  • interpolate the color values from the neighboring pixel in the corresponding color If necessary, continuity information from the pixels of other color values is also considered and used.
  • Preferred color cameras are e.g. 3-chip color cameras that directly measure an intensity value for red, green and blue for each pixel. These preferred color cameras should therefore be cameras which directly measure the color values required for each pixel.
  • the color information can be linked, preferably, for example via methods of computer learning, between the two samples.
  • this linking takes place separately for each class of cells in order to capture specific staining behavior as well as possible.
  • the 5 main groups who differentiated in the 5 Part differential diagnosis who train together. It may then be advantageous to still be within such a group in a second step to learn the colorations of specific features in order to obtain as good and comprehensive color contrasting as possible.
  • the 3D information e.g. in the form of topographic information or as volumetric or tomographic information.
  • this 3D information and information advantageously derived therefrom e.g. an effective 3D profile, both for the uncolored image and preferably for the colored image.
  • all methods known from computer learning are in principle applicable, such as e.g. PLS, PLSDA, PCA or neural networks (CNN) or deep CNNs.
  • each class of cells to be learned and / or group of cells learned together the classification takes place in at least as many color values as on the original sample under divisible and / or can be displayed on the display device.
  • color spaces are common for color representation, where for each pixel per color e.g. 256 color values, ie 8 bits or 1 byte color depth, or 65536 color values, ie 16 bits or 2 bytes or 1 double byte can be used.
  • a great advantage of digital dyeing according to the invention is that samples can now be vergli chen between regions, which typically with other coloring Tocolepro and thus other color effect of the cells work ar. If an undyed sample to be examined has been taken up, this can optionally be dyed according to the invention with different dyed color structures which correspond to the respective regional dyeing protocol. On the one hand, this offers the advantage that it is now possible to use the preferred or advantageous staining protocol for different pathologies. On the other hand, a doctor may ask another doctor for a explicatatory opinion, as he can now show him the picture in the familiar color. Digital staining can circumvent the hurdles created by the staining protocols, because now the doctor can choose his customary staining to carry out the diagnosis. So then also, e.g. Doctors from Europe and the USA work better together for the benefit of the patient. This allows for a worldwide collaboration of doctors and / or hematologists in terms of telemedicine.
  • every doctor worldwide can contribute to a secure diagnosis.
  • a database in particular for rare pathological cases of observed and / or measured cells over the world wide networking with telemedicine advantageously one
  • a well-grounded "ground truth" dataset which is a combination of image and / or 3D information and the findings backed up by at least two physicians, is an important component of this procedure, as well in the possibility according to the invention in the digital dataset for each cell taken Separate and adapted to be able to change the focus of the images without the doctor finding himself sitting on a microscope.
  • This data set which is independently diagnosed by several physicians, can then, assuming a match in the findings, e.g. be used to further train computer models, which can then be played back on the globally installed devices, in order to further develop each device in the recognition quality for certain cells, or in the ability to safely identify special pathologies automatically.
  • the analyzer may also be advantageous for the analyzer according to the invention to take pictures and then store them, e.g. in a cloud-based application, or more generally to another computing unit, where then the recognition of the cells, their segmentation and an automated evaluation, e.g. in the sense of an association with a class of cell type or of a diagnosis as a recognition of certain disease images and / or the determination of a suspicion on a specific clinical picture takes place.
  • the color representation is so adjusted that the doctor can assume a color-fast color rendering. This is particularly important in hematology because small color differences and structural differences in the image of the cell can give an indication of conspicuousness, especially in the case of pathological or suspicious cells.
  • the color reproduction in particular special with respect to the stable color reproduction such as the DICOM standard for medical devices (dicom.nema.org/).
  • the user and / or user can then focus on the software through the cell images and / or the flat image from the slide or from the flowcell by operation on a software.
  • a software for a correspondingly secure, or possibly pre given, data connection to the data on a local data memory, a superordinate data memory, e.g. in a hospital, and / or a cloud. Preference may be given to e.g. a consultation, the data will be made directly accessible to a doctor.
  • a 3D-capable display is used, which can be viewed either with or more preferably without further optical Hilfsmit tel and generated for the user a 3D He heintik.
  • the 3D capable display is part of a computer and / or a mobile terminal, e.g. a tray.
  • the 3D effect can be switched on and off.
  • Data glasses also referred to as smart glasses or VR glasses, are preferably used for the 3D representation of the data set.
  • a working mode master-slave is preferably provided for a remote diagnosis in telemedicine, if simultaneous diagnosis of the images by several Doctors, for example, in a medical conference, as a Mas ter navigation over the images of the cells and / or from the slide and / or the flow cell (Flowcell) leads and the other doctors then this can follow without own controlling the system.
  • each of the doctors can also focus on his image independently of the others and / or color the image according to his usual staining protocol.
  • 1 shows an automatic analyzer for analyzing cells in a sample.
  • the automatic analyzer shown in FIG. 1 for analyzing cells in a sample comprises an optical microscope (1) comprising a light source (4) for illuminating a sample (2) and a converging lens for collecting and focusing light rays (6) generated by go out of the illuminated sample (2).
  • the sample (2) is a blood sample containing blood cells (3).
  • the sample (2) is located in a microfluidic flow cell (10).
  • the microscope (1) comprises a light field camera (8) comprising a digita les recording device, the recording device comprising egg nen CCD chip or a CMOS chip, for receiving the micro-scope (1) imaged light field.
  • the microscope (1) comprises another camera for taking an image in the object plane, which is designed as a high-resolution 3-chip color camera (9).
  • the lateral resolution of the color camera (9) be four times the effective lateral resolution of the light field camera (8).
  • the focus of the color camera (9) is on one Set the central range of the measuring range of the light field camera (8), for example to a virtual depth in the range of 3 to 5.
  • the incoherent illumination sigma on the microscope (1) is 1.0.
  • the microscope (1) is a micro-scope that can also be operated as a differential interference contrast (DIC) microscope (1).
  • DIC differential interference contrast

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Abstract

L'invention concerne un analyseur permettant d'analyser un échantillon médical, ledit analyseur comprenant : un microscope optique pour représenter un champ lumineux dans une zone d'objet destinée à la représentation de l'échantillon, le microscope comprenant une source lumineuse pour éclairer l'échantillon et un objectif comprenant une lentille convergente pour collecter et focaliser des faisceaux lumineux à partir de l'échantillon éclairé. L'invention concerne également un appareil d'enregistrement numérique pour enregistrer les faisceaux lumineux, le microscope étant pourvu d'une caméra à champ lumineux pour recevoir le champ lumineux représenté dans le microscope à partir de la zone d'objet.
EP19728133.0A 2018-05-30 2019-05-16 Analyseur destiné à l'analyse tridimensionnelle d'un échantillon médical au moyen d'une caméra à champ lumineux Pending EP3803495A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
EP18000484.8A EP3575848A1 (fr) 2018-05-30 2018-05-30 Analyseur destiné à l'analyse tridimensionnelle d'un échantillon médical au moyen d'un appareil photographique plénoptique
PCT/IB2019/054051 WO2019229573A1 (fr) 2018-05-30 2019-05-16 Analyseur destiné à l'analyse tridimensionnelle d'un échantillon médical au moyen d'une caméra à champ lumineux

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EP3803495A1 true EP3803495A1 (fr) 2021-04-14

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JP2021525866A (ja) 2021-09-27
CN112236705A (zh) 2021-01-15
US20210217190A1 (en) 2021-07-15
WO2019229573A1 (fr) 2019-12-05

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