METHOD FOR DIGITALLY STAINING CELLS

Abstract

The invention relates to a method for digitally staining a cell and/or a medical preparation, the method comprising the following steps: determining three-dimensional information of a cell and/or of a medical preparation by means of an analyser for analysing a medical sample, the analyser comprising an apparatus for determining the three-dimensional information of the cell and/or of the medical preparation; digitally staining the cell and/or the medical preparation according to a predetermined correlation between the three-dimensional information of the cell and/or of the medical preparation and the staining of a corresponding cell and/or medical preparation and/or cellular and/or sub-cellular structures of the cell and/or of the medical preparation by means of a staining protocol; representing the digitally stained cell and/or the preparation, the representation involving a predetermined defocus region, and regions of the cell and/or of the preparation being represented by means of different digital staining in the area of the defocus region as corresponding modulations of colour intensities and/or as mixed colour.

Claims

1. A method for digitally staining a cell or medical preparation, the method comprising the steps of: a) ascertaining three-dimensional information about a cell or medical preparation using an analyzer for analyzing a medical sample, the analyzer comprising an apparatus for ascertaining the three-dimensional information about the cell or medical preparation, b) digitally staining the cell or medical preparation in accordance with a predetermined assignment between the three-dimensional information about the cell or medical preparation and the staining of a corresponding cell or medical preparation or cellular or subcellular structures of the cell or medical preparation using a staining protocol, and c) representing the digitally stained cell or preparation, where the representation takes place with a predetermined defocus range, and where regions of the cell or preparation with different digital staining in the region of the defocus range are represented as corresponding modulations of color intensities or as mixed color.

2. The method as claimed in claim 1, wherein the representing in step c) comprises a displaying or storage of a two-dimensional image of the digitally stained cell or preparation, where the displaying of the two-dimensional image corresponds to the image in a selected focal plane of the cell or preparation and where mutually superposed regions of the cell or preparation with different digital staining in the region of the defocus range are displayed as corresponding modulations of color intensities or as mixed color.

3. The method as claimed in claim 1, wherein the three-dimensional information ascertained in step a) comprises information about the sample at cellular or subcellular orders of magnitude.

4. The method as claimed in claim 3, wherein step a) comprises segmentation of the ascertained three-dimensional information for recognition of cellular or subcellular structures of the sample.

5. The method as claimed in claim 1, wherein the three-dimensional information ascertained in step a) is three-dimensional image information.

6. The method as claimed in claim 1, wherein the three-dimensional information ascertained in step a) comprises holographic information.

7. The method as claimed in claim 1, wherein the analyzer for ascertaining the three-dimensional information in step a) comprises an optical microscope for imaging a light field in an object region for imaging the sample, the microscope comprising a light source for illuminating the sample and an objective comprising a converging lens for converging and focusing light beams emanating from the illuminated sample, and a digital recording device for recording the light beams, where a light field camera for recording the light field from the object region that is imaged in the microscope is provided on the microscope, where the light field camera comprises the digital recording device.

8. The method as claimed in claim 7, wherein the light field camera comprises a microlens array with lenses of different focal lengths, where the microlens array is able to image an intermediate image of the light field imaged in the microscope onto the digital recording device.

9. The method as claimed in claim 1, wherein the analyzer is an automated hematology analyzer, and where the cell is a blood cell.

10. The method as claimed in claim 7, wherein the microscope is an amplitude contrast microscope or a phase contrast microscope or a differential interference contrast (DIC) microscope.

11. The method as claimed in claim 7, wherein a further camera for recording an image in an object plane in the object region is provided on the microscope, where the further camera has a lateral resolution which is higher than the lateral resolution of the light field camera and where the further camera has a larger field of view than the light field camera.

12. The method as claimed in claim 11, wherein the further camera is a color camera.

13. The method as claimed in claim 7, wherein an incoherent illumination sigma on the microscope is greater than 0.8, where sigma is given as the ratio of the numerical aperture of the illumination of the sample to the numerical aperture of the objective, and where the microscope is a differential interference contrast (DIC) microscope.

14. The method as claimed in claim 1, wherein the sample in step a) is supplied to the analyzer via a flow cell and where the object plane of the microscope lies in the flow cell, or wherein the sample in step a) is supplied to the analyzer on an object carrier via a sample supply facility for object carriers.

15. The method as claimed in claim 1, wherein the cells or medical preparations are not stained.

16. The method as claimed in claim 1, further comprising the step of: d) assigning the cell to a cell type on the basis of predetermined information and the three-dimensional information about the cell ascertained in step a).

17. The method as claimed in claim 1, wherein the staining protocol is the May-Grünwald-Giemsa staining protocol, modified Wright staining, Wright-Giemsa staining or Romanowsky staining or is a predetermined color table which does not correspond to a chemical staining protocol.

18. A method for training a method for digitally staining a cell or medical preparation, the method comprising the steps of: aa) ascertaining three-dimensional information about a cell or medical preparation using a first analyzer for analyzing a medical sample, the first analyzer comprising an apparatus for ascertaining the three-dimensional information about the cell or medical preparation, bb) coloring the cell or medical preparation by implementing a staining protocol, cc) ascertaining three-dimensional information about the cell or medical preparation using a second analyzer for analyzing a medical sample, the second analyzer comprising an apparatus for ascertaining the three-dimensional information about the cell or medical preparation comprising a color contrast, where the color contrast is sensitive to the coloration of the staining protocol, and dd) compiling and storing an assignment between the three-dimensional information about the cell or medical preparation ascertained in step aa) and step cc).

19. The method as claimed in claim 18, wherein step cc) comprises segmentation of the ascertained three-dimensional information comprising the color contrast for recognizing cellular or subcellular structures of the sample.

20. The method as claimed in claim 18, wherein the compiling of the assignment in step dd) comprises applying techniques of machine learning or artificial intelligence using a computer.

21. The method as claimed in claim 18, wherein the first and second analyzers are the same device.

22. The method for digitally staining a cell or medical preparation as claimed in claim 1, wherein the assignment in step b) has been determined by a method for training a method for digital staining of a cell or medical preparation comprising: aa) ascertaining three-dimensional information about a cell or medical preparation using a first analyzer for analyzing a medical sample, the first analyzer comprising an apparatus for ascertaining the three-dimensional information about the cell or medical preparation, bb) coloring the cell or medical preparation by implementing a staining protocol, cc) ascertaining three-dimensional information about the cell or medical preparation using a second analyzer for analyzing a medical sample, the second analyzer comprising an apparatus for ascertaining the three-dimensional information about the cell or medical preparation comprising a color contrast, where the color contrast is sensitive to the coloration of the staining protocol, and dd) compiling and storing an assignment between the three-dimensional information about the cell or medical preparation ascertained in step aa) and step cc).

Description

BRIEF DESCRIPTION OF THE DRAWING

[0231] The analyzer of the invention is elucidated in more detail by means of a specific exemplary embodiment, on the basis of the appended drawing. The example shown represents one preferred embodiment of the invention. In the drawing:

[0232] FIG. 1 shows an automated analyzer for analyzing cells in a sample.

DETAILED DESCRIPTION OF THE INVENTION

[0233] The automated 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 converging and focusing light beams (6) which emanate from 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) further comprises a light field camera (8) comprising a digital recording device, the recording device comprising a CCD chip or a CMOS chip, for recording the light field imaged in the microscope (1). The microscope (1) further comprises an additional camera for recording an image in the object plane, this camera being embodied as a high-resolution three-chip color camera (9). The lateral resolution of the color camera (9) is four times the effective lateral resolution of the light field camera (8). The focus of the color camera (9) is set on a central region of the measuring range of the light field camera (8), e.g., to a virtual depth in the range from 3 to 5. The incoherent illumination sigma on the microscope (1) is 1.0. The microscope (1) is a microscope which may also be operated as a differential interference contrast (DIC) microscope (1).

LIST OF REFERENCE SYMBOLS

[0234] 1 Microscope [0235] 2 Sample [0236] 3 Blood cell [0237] 4 Light source [0238] 6 Light beams [0239] 8 Light field camera [0240] 9 Color camera [0241] 10 Flow cell