IMAGE FORMING CYTOMETER

• US 20190323944A1
• Filed: 07/03/2019
• Published: 10/24/2019
• Est. Priority Date: 05/28/2013
• Status: Active Grant

• Current Assignee: ChemoMetec A/S
• Original Assignee: ChemoMetec A/S
• Inventors: Glensbjerg, Martin, Holm, Johan, Kjaerulff, Soren, Ravn Hansen, Frans Ejner
• Granted Patent: US 10,921,234 B2
• Time in Patent Office: Days
• CPC Class Codes: G01N 15/01 specially adapted for biological cells, e.g. blood cells investigating sedimentation of particle suspensions in blood G01N15/05
• CPC Class Codes: G01N 15/0227 using imaging; using holography
• CPC Class Codes: G01N 15/1433 using image recognition
• CPC Class Codes: G01N 15/1434 Optical arrangements
• CPC Class Codes: G01N 2015/1006 for cytology
• CPC Class Codes: G01N 2015/1438 Using two lasers in succession
• CPC Class Codes: G01N 2015/144 Imaging characterised by its optical setup
• CPC Class Codes: G01N 2015/1493 Particle size
• CPC Class Codes: G01N 2015/1495 Deformation of particles
• CPC Class Codes: G06T 2207/10056 Microscopic image
• CPC Class Codes: G06T 2207/10064 Fluorescence image
• CPC Class Codes: G06T 2207/30024 Cell structures in vitro; Tissue sections in vitro
• CPC Class Codes: G06T 7/0012 Biomedical image inspection
• CPC Class Codes: G06V 20/69 Microscopic objects, e.g. biological cells or cellular parts

Abstract

The present invention relates to methods and systems for image cytometry analysis, typically at low optical magnification, where analysis is based on detection of biological particles using UV bright field and optionally one or more sources of excitation light.

Claims

1. An imaging apparatus, comprising:

a first light source configured for emitting light into a sample region;

a collimator for forming collimated light from the first light source and directing the collimated light along an optical axis of the imaging apparatus and into the sample region;

a second light source comprising a first excitation light source configured for emitting excitation light into the sample region;

an image forming element for forming at least one image of at least part of the sample region on an array of detection elements;

wherein the sample region is located between the collimator and the array of detection elements;

wherein the imaging apparatus is configured to be interchanged between a bright field mode, a dark field mode and a fluorescence mode.

2. The imaging apparatus according to claim 1, wherein the first light source provides the light for the bright field mode and the dark field mode.

3. The imaging apparatus according to claim 1, further comprising a modulator located between the sample region and the array of detection elements, the modulator operable to interchange between forming the bright-field image and the dark-field image.

4. The imaging apparatus according to claim 3, wherein the modulator comprises an aperture configured for attenuating light during the formation of the bright-field image, wherein the light is passing the sample region as collimated light.

5. The imaging apparatus according to claim 3, wherein the modulator comprises an obstruction configured for attenuating light leaving the sample region as collimated light during the formation of the dark-field image.

6. The imaging apparatus according to claim 5, wherein the obstruction comprises an at least partly opaque centre surrounded by an at least partly transparent area.

7. The imaging apparatus according to claim 3, wherein the image forming element is located between the sample region and the array of detection elements, and the modulator is located at or close to the focal plane of the image forming element.

8. The imaging apparatus according to claim 6, wherein the at least partly opaque centre is located in or close to a focal point of the image forming element.

9. The imaging apparatus according to claim 3, wherein the modulator comprises a phase contrast microscopy modulator.

10. The imaging apparatus according claim 1, wherein the excitation light is at an incidence angle between 10 and 80 degrees, or of 90 degrees, or between 110 and 180 degrees relative to the optical axis so as to provide the fluorescence image.

11. The imaging apparatus according to claim 1, wherein the first and/or second light source(s) is/are configured for emitting light in duration between 0.0001 and 0.1000 second.

12. The imaging apparatus according to claim 1, wherein the first and/or second light source(s) is/are configured for emitting light in duration for more than a 1 second.

13. The imaging apparatus according to claim 1, wherein the light source(s) is/are optically connected to an optical means configured for providing light with a substantially uniform intensity across the sample region and/or across a region imaged by the array of detection elements.

14. The imaging apparatus according to claim 13, wherein the optical means comprises an array of micro lenses.

15. The imaging apparatus according claim 13, wherein the optical means comprise two arrays of cylindrical micro lenses, wherein the lenses in one of the arrays are orientated with a major axis being perpendicular to a major axis of the lenses of the other array.

16. The imaging apparatus according to claim 1, wherein the image forming element is configured for transmitting light in the wavelength region of between 200 nm and 1000 nm.

17. The imaging apparatus according to claim 1, wherein the sample region is adapted to hold a liquid sample with biological particles.

18. The imaging apparatus according to claim 1, wherein the image forming element is an objective and/or one or more lenses and/or one or more curved mirrors.

19. The imaging apparatus according to claim 1, wherein the image forming element is configured for providing a linear enlargement of the sample.

20. The imaging apparatus according to claim 19, wherein the linear enlargement is smaller than 20:

1.

21. The imaging apparatus according to claim 1, wherein the first light source and/or the second light source is/are a light emitting diode (LED) and/or a diode laser and/or laser.

22. A method for the assessment of at least one quantity parameter and/or one quality parameter of a biological sample, comprising:

applying a volume of the biological sample to a sample compartment having parallel wall parts defining an exposing area, the wall parts allowing light from an imaging apparatus to pass through the wall parts of the sample compartment, wherein the imaging apparatus comprises a first light source configured for emitting light into the sample compartment and a second light source comprising a first excitation light source configured for emitting excitation light into the sample compartment;

illuminating the sample compartment with collimated light from the first light source, and exposing, onto a 2-dimensional array of active detection elements, light having passed through the sample compartment, thus recording a bright-field image of spatial light intensity information;

illuminating the sample compartment with collimated light from the first light source, and exposing, onto a 2-dimensional array of active detection elements, light having interacted with the biological sample, thus recording a dark-field image of spatial light intensity information;

illuminating the sample compartment with excitation light from the first excitation light source, and exposing, onto the 2-dimensional array of active detection elements, fluorescent light having passed through the sample compartment, thus recording a fluorescent image of spatial light intensity information;

processing all three images in such a manner that light intensity information from individual biological objects are identified as distinct from light intensity information from the background; and

correlating the results of the processing to the at least one quantity parameter and/or quality parameter of biological particles in the biological sample.

23. The method according to claim 22, wherein two or more images of spatial light intensity information are recorded, each of the images being recorded with two or more substantially different modulators.

24. The method according to claim 22, wherein the parameter to be assessed is location of biological particles in the spatial light intensity image.

25. The method according to claim 24, wherein the location of biological particles in the first spatial light intensity image is used to determine presence of light intensity information in another recorded image of light intensity information associated to biological particles, where the other light intensity information is fluorescence.

26. The method according to claim 24, wherein the location of biological particles is determined by combining information in a first and a second image of spatial light intensity information, where the images are recorded using illumination from the first light source and applying substantially different modulators for the images.

27. The method according to claim 26, wherein the images are dark-field and bright-field images.