Method and a system for determination of somatic cells in milk

• US 6,731,100 B1
• Filed: 11/01/1999
• Issued: 05/04/2004
• Est. Priority Date: 05/05/1997
• Status: Expired due to Term

• Current Assignee: ChemoMetec A/S
• Original Assignee: ChemoMetec A/S
• Inventors: Jeppesen, Jesper Myron, Glensbjerg, Martin, Hansen, Frans Ejner Rvan, Arnvidarson, Borkur
• Primary Examiner(s): Cuneo, Kamand
• Assistant Examiner(s): PATEL, PARESH H
• Application Number:
  US09/403,959
• Time in Patent Office: 1,646 Days
• Field of Search: 324/71.4, 324/638, 324/637, 324/639, 356/72, 356/336
• US Class Current: 324/71.4
• CPC Class Codes: G01N 15/1433 using image recognition
• CPC Class Codes: G01N 2015/1486 Counting the particles
• CPC Class Codes: G01N 21/51 inside a container, e.g. in an ampoule G01N21/53 takes precedence
• CPC Class Codes: G01N 21/59 Transmissivity G01N21/25 takes precedence
• CPC Class Codes: G01N 21/6486 Measuring fluorescence of biological material, e.g. DNA, RNA, cells G01N21/6428 takes precedence
• CPC Class Codes: G01N 33/04 Dairy products

Abstract

Assessment of the number of somatic cells in a liquid milk or milk product analyte material comprises arranging a volume of a liquid sample representing the analyte material in a sample compartment and allowing electromagnetic signals from the sample in the compartment to pass through and be exposed to the exterior. There is exposed onto an array of active detection elements, an at least one-dimensional spatial representation of electromagnetic signals from the sample. The representation is detectable as an intensity by individual active detection elements. Representations of electromagnetic signs from the somatic cells are identified. The size of the volume of the liquid sample is sufficiently large to permit the assessment of the number of somatic cells to fulfill a predetermined requirement to the statistically quality of the assessment based on substantially one exposure.

Claims

1. A method for the assessment of the number of somatic cells in a liquid milk or milk product analyte material, comprising

arranging a volume of a liquid sample representing the analyte material in a sample compartment having a wall part defining an exposing area, the wall part allowing electromagnetic signals from the sample in the compartment to pass through the wall and to be exposed to the exterior,

exposing, onto an array of active detection elements, an at least one dimensional spatial representation of said electromagnetic signals having passed through the wall part from the sample in the sample compartment,

detecting the representation as intensities by individual active detection elements,

processing the intensities in order to identify representations of electromagnetic signals from the somatic cells as distinct from representations of electromagnetic signals from background,

correlating the results of the processing to the number of somatic cells in the liquid analyte material, and

the size of the volume of the liquid sample being more than 0.01 μ

l and the cells therein being identified by one exposure, that the assessment of the number of somatic cells fulfills a predetermined requirement to the statistical quality of the assessment based on substantially one exposure.

2. The method according to claim 1, wherein the representation of the electromagnetic signals is a one-dimensional image representation.

3. The method according to claim 1, wherein the spatial representation of the electromagnetic signals is a two-dimensional image representation.

4. The method according to claim 1, wherein the array of detection elements is arranged in such a way that the series of detection elements form a straight line.

5. The method according to claim 4, wherein the array of detection elements is arranged in two directions in such a way that the detection elements form a series of parallel straight lines, the series forming a rectangle.

6. The method according to claim 1, wherein the exposure of the spatial representation of electromagnetic signals onto the array of detection elements is performed by focusing an image of electromagnetic signals from at least a part of the exposing area onto the array of detection elements by means of a focusing means.

7. The method according to claim 6, wherein the focusing means is a lens comprising of one or several elements.

8. The method according to claim 1, wherein the spatial representation exposed onto the array of detection elements is subject to such a linear enlargement that the ratio of the image of a linear dimension on the array of detection elements to the original linear dimension in the exposing area is between 3:

1 and 1;

100.

9. The method according to claim 1, wherein the individual somatic cells particles to be assessed are imaged on at the most 5 detection elements.

10. The method according to claim 1, wherein the interior of the sample compartment has an average thickness of between 20 μ

m and 200 μ

m.

11. The method according to claim 1, wherein the sample compartment has dimensions, in a direction parallel to the array of detection elements, in the range between 1 mm by 1 mm and 10 mm by 10 mm.

12. The method according to claim 1, wherein the volume of the liquid sample from which electromagnetic radiation is exposed onto the array is in the range between 0.04 μ

l and 4 μ

l.

13. The method according to claim 1, wherein the sample in the sample compartment is at stand still during the exposure.

14. The method according to claim 1, wherein the sample in the sample compartment is moved through the sample compartment during the exposure, and the exposure is performed over a short period of time to substantially obtain stand still condition during the exposure.

15. The method according to claim 1, wherein at least a major part of the electromagnetic radiation emitted from the sample during exposure originates from or is caused by electromagnetic radiation supplied to the sample from a light source, at least a major part of the radiation from the light source having a direction which is transverse to the wall of the sample compartment.

16. The method according to claim 1, wherein the size of the volume of the liquid sample allows identification therein of at least four somatic cells.

17. The method according to claim 1, wherein arranging a volume of a liquid sample comprises arranging a volume of 0.04 μ

or more of the liquid sample representing the analyte material in the sample compartment, and

wherein processing the intensities comprises using an enlargement so that the ratio of the image of a linear dimension on the array of detection elements to the original linear dimension in the exposing area is between 3;

1 and 1;

100, and such that individual somatic cells are imaged on at the most 25 detection elements of the array of detection elements, and

wherein the sample in the sample compartment being at stand still or substantially at stand still during the exposure, and at least a major part of the electromagnetic radiation emitted from the sample during exposure originates from or is caused by electromagnetic radiation supplied to the sample from a light source at least a major part of the radiation from which has a direction which is transverse to the wall of the sample compartment.

18. The method according to claim 1, wherein the signal which is detected by the detecting elements originates from one or more molecules which bind to, are retained within, or interact with, the biological particles, such molecules being added to the sample or the isolated particles before or during exposure, the molecules being molecules giving rise to one or several of the following phenomena:

attenuation of electromagnetic radiation, photoluminescence when illuminated with electromagnetic radiation, scatter of electromagnetic radiation, raman scatter.

19. The method according to claim 18, wherein an effective amount of one or more nucleic acid dyes and/or one or more potentiometric membrane dyes is added.

20. The method according to claim 19, wherein a nucleic acid dye or nucleic acid dyes is/are added in an amount of 0.3-30 μ

g per ml of the sample.

21. The method according to claim 19, wherein the one or more nucleic acid dyes are selected from the group consisting of:

phenanthridines, acridine dyes, cyanine dyes, indoles and imidazoles.

22. The method according to claim 1, wherein the duration of the exposure is in the range from 100 milliseconds to 5 seconds.

23. The method according to claim 22, wherein the duration of the exposure is in the range of 0.5 to 3 seconds.

24. The method according to claim 22, wherein the exposure is performed as a single exposure.

25. The method according to claim 1, wherein the sample compartment is part of a disposable unit.

26. The method according to claim 1, wherein the assessment is performed in an automated system at a rate of at least 300 assessments per hour.

27. The method according to claim 1, wherein the assessment is performed substantially simultaneously with milking.

28. The method according to claim 27, wherein the assessment is performed at-line.

29. The method according to claim 27, wherein the sample is a milk sample collected during milking.

30. The method according to claim 27, wherein the results of the assessments are transferred to one or several information storage means, the information storage means also being able to store other information about the milking.

31. The method according to claim 30, wherein the information storage means includes means to indicate whether the milk being milked should be directed to one or several of storage facilities or outlets, the indication being based on the assessment of the number of somatic cells per volume, as well as on other information present in the information storage means about milking of individual animals or bulk tank milk, the other information being selected from:

conductivity, impedance, temperature, fat content, protein content, lactose content, urea content, citric acid content, ketone content, somatic cell count.

32. The method according to claim 31, wherein the purpose of the direction of any milk being milked to one or several of storage facilities or outlets is to adjust the properties of any bulk of milk with regard to the number of somatic cells per volume by mixing the bulk of milk with milk having a lower somatic cell count, and/or by excluding milk having a somatic cell count exceeding a predetermined limit.

33. The method according to claim 1, wherein the assessment is carried out after the milking has taken place, the milk not being altered before measurement.

34. The method according to claim 1, wherein the assessment is carried out after the milking has taken place, the milk being modified before measurement, the modification being one or several of addition of one or more chemical components which substantially inhibit bacterial growth in the sample material, addition of one or more chemical components which substantially inhibit the growth of fungus, addition of one or more chemical components which have colouring properties, said colouring being used to aid visual identification of the milk.

35. The method according to claim 1, wherein the assessment is carried out simultaneously with the assessment of the amount of any constituent in said sample material, the constituent being selected from:

fat, protein, lactose, urea, citric acid, glucose, ketones, carbon dioxide, oxygen, pH, potassium, calcium, sodium.

36. The method according to claim 35, wherein the assessment of any chemical constituent is based on spectrophotometric measurement selected from:

mid-infrared attenuation, near-infrared attenuation, visible attenuation, ultra-violet attenuation, photoluminescence, raman scatter, nuclear magnetic resonance.

37. The method according to claim 35, wherein the assessment of any chemical constituent is based on potentiometric measurement.

38. The method according to claim 1, wherein the sample material is a milk sample taken from one quarter of the udder.

39. The method according to claim 1, wherein the somatic cells or fragments thereof are determined, and the sample material is a milk sample, the sample of the sample material is illuminated in the sample compartment with electromagnetic radiation where at least a portion of said electromagnetic radiation has energy which can give rise to a photoluminescence signal, the signal originating at least from said somatic cells or portions of said somatic cells or components interacting with or bound to the somatic cells or portions thereof.

40. The method according to claim 39, wherein the signal originates from one or several types of molecules intentionally added to said sample which bind to or interact with the somatic cells or parts of the somatic cells.

41. The method according to claim 1, wherein somatic cells or fragments thereof are determined, and the sample material is a milk sample, the purpose of the assessment being to obtain information about the health status of a milking animal, the sample of the sample material is placed in a sample compartment by the use of a flow means capable of replacing the sample within the sample compartment with a different sample, the sample of the sample material is illuminated in the sample compartment with electromagnetic radiation where at least a portion of said electromagnetic radiation has energy which can give rise to a photoluminescence signal, the signal originating at least from said somatic cells or portions of said somatic cells or components interacting with or bound to the somatic cells or portions thereof.

42. The method according to claim 1, wherein somatic cells or fragments thereof are determined, and the sample material is a milk sample, the sample of the sample material is placed in a sample compartment by the use of a flow means capable of replacing the sample within the sample compartment with a different sample the time between the replacement of sample material being shorter than 30 seconds, the sample of the sample material is illuminated in the sample compartment with electromagnetic radiation where at least a portion of said electromagnetic radiation has energy which can give rise to a photoluminescence signal, the signal originating at least from said somatic cells or portions of said somatic cells or components interacting with or bound to the somatic cells or portions thereof.

43. The method according to claim 42, wherein the signal originates from one or several types of molecules intentionally added to said sample which interact or bind to or interact with the somatic cells or parts of the somatic cells.

44. The method according to claim 1, wherein somatic cells or fragments thereof are determined, and the sample material is a milk sample, the assessment being performed at the beginning of milking, or during milking, or immediately after milking has taken place, the sample of the sample material is placed in a sample compartment by the use of a flow means capable of replacing the sample within the sample compartment with a different sample flowing milk directly from a milking unit or flowing milk from an intermediate reservoir which is gradually filled during milking, the sample of the sample material is illuminated in the sample compartment with electromagnetic radiation where at least a portion of said electromagnetic radiation has energy which can give rise to a photoluminescence signal, the signal originating at least from said somatic cells or portions of said somatic cells or components interacting with or bound to the somatic cells or portions thereof.

45. The method according to claim 44, wherein the signal originates from one or several types of molecules intentionally added to said sample which interact or bind to or interact with the somatic cells or parts of the somatic cells.

46. The method according to claim 1, wherein somatic cells or fragments thereof are determined, and the sample material is a milk sample, a portion of the sample material is placed in a sample compartment being at least a part of a unit which can be replaced between every assessment or where each of said units can only be used for said assessment of one of said sample materials, the sample of the sample material is illuminated in the sample compartment with electromagnetic radiation where at least a portion of said electromagnetic radiation has energy which can give rise to a photoluminescence signal, the signal originating at least from said somatic cells or portions of said somatic cells or components interacting with or bound to the somatic cells or portions thereof.

47. The method according to claim 46, wherein the signal originates from one or several types of molecules intentionally added to said sample which interact or bind to or interact with the somatic cells or parts of the somatic cells.

48. The method according to claim 1, wherein the sample is milk which is undiluted, except for the addition of the reagents used in the assessment, the reagents being non-aqueous.