Apparatus and methods for microscopic analysis of biological samples

Abstract

An apparatus (20) and method for analyzing a blood sample are provided. The blood sample is mixed with a fluorescently labeled antibody and placed in a sample chamber (52). The sample chamber containing the blood sample is placed in an optical measurement unit (31). One or more fluorescence microscope images of the blood sample in the sample chamber are acquired using the microscope of the optical measurement unit. A computer processor (28) performs an immunophenotyping of the blood sample by analyzing one or more fluorescence microscope images of the blood sample and generates an output at least partially accordingly. Other applications are also described.

Claims

1. The preparation of blood samples for analysis, Mixing the fluorescently labeled antibody with the blood sample, The blood sample is placed inside the sample chamber, The blood sample is prepared for analysis by placing the sample chamber containing the blood sample inside within the optical measurement unit. Using the microscope of the optical measurement unit, one or more fluorescence microscope images of the blood sample in the sample chamber are acquired. Using a computer processor, By analyzing one or more fluorescence microscope images of the blood sample, an immunophenotypic test is performed on the blood sample. To generate output in accordance with this, at least partially. A method that includes this.

2. The method according to claim 1, further comprising: counting entities in the blood sample by analyzing one or more microscopic images of the blood sample; determining the characteristics of the blood sample based on the counting of entities in the sample and the immunophenotyping test, wherein generating the output includes generating the output in accordance with the determination of the characteristics of the blood sample.

3. The method according to claim 1, wherein mixing the fluorescently labeled antibody with the blood sample includes mixing a fluorescently labeled antibody against one or more cell surface markers listed in Table 1, and performing an immunophenotypic test of the blood sample by analyzing one or more fluorescence microscope images of the blood sample includes identifying an index corresponding to the one or more cell surface markers listed in Table 1.

4. The method according to claim 1, wherein performing an immunophenotypic test of the blood sample by analyzing one or more fluorescence microscope images of the blood sample includes determining the average fluorescence intensity level of cells bound to the fluorescently labeled antibody in one or more of the microscope images of the blood sample.

5. The method according to claim 1, wherein mixing the fluorescently labeled antibody with the blood sample comprises mixing a fluorescently labeled antibody against at least one of CD14 and CD16.

6. The method according to claim 1, wherein placing the blood sample into the sample chamber includes placing the blood sample into the sample chamber in an unwashed state.

7. The method according to claim 1, wherein mixing the fluorescently labeled antibody with the blood sample comprises mixing with an anti-CD3 fluorescently labeled antibody, and performing an immunophenotypic test of the blood sample by analyzing one or more fluorescence microscope images of the blood sample comprises identifying T cell lymphocytes in the blood sample.

8. The method according to claim 1, wherein mixing the fluorescently labeled antibody with the blood sample includes mixing with an anti-CD3 fluorescently labeled antibody, and performing an immunophenotypic test of the blood sample by analyzing one or more fluorescence microscope images of the blood sample includes counting T-cell lymphocytes in the blood sample.

9. The method according to claim 1, wherein mixing the fluorescently labeled antibody with the blood sample includes mixing with an anti-CD19 fluorescently labeled antibody, and performing an immunophenotypic test of the blood sample by analyzing one or more fluorescence microscope images of the blood sample includes identifying B cell lymphocytes in the blood sample.

10. The method according to any one of claims 1 or 3 to 9, wherein mixing the fluorescently labeled antibody with the blood sample comprises mixing an anti-CD64 fluorescently labeled antibody, and generating the output comprises generating an output that, at least in part, based on the immunophenotyping test, the subject from whom the blood sample was taken is suspected to have an infectious disease.

11. The method according to claim 10, wherein generating the output includes generating an output that, at least in part, based on the immunophenotypic test, the subject from whom the blood sample was taken is suspected to have sepsis.

12. The method further includes identifying one or more biomarkers selected from the group consisting of procalcitonin (PCT), C-reactive protein (CRP), interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-10 (IL-10), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), presepsin, thrombomodulin, lactate-soluble urokinase-type plasminogen-activating factor receptor (suPAR), soluble trigger receptor-1 (sTREM-1) expressed in bone marrow cells, lipopolysaccharide-binding protein (LBP), N-terminal pro-brain natriuretic peptide (NT-proBNP), neutrophil gelatinase-binding lipocalin (NGAL), adrenomedullin, resistin, pro-adrenomedullin (MR-proADM), D-dimer, and pancreatic stone protein (PSP). The method of claim 10, wherein generating the output indicating that the subject from which the sample was taken is suspected to be suffering from an infectious disease comprises generating the output indicating that the subject from which the sample was taken is suspected to be suffering from an infectious disease based on the immunophenotyping test and one or more biomarkers selected from the group.

13. This further includes receiving the results of one or more diagnostic tests selected from the group consisting of vital sign measurement, blood culture, urine culture, chest X-ray, CT scan, ultrasound, lumbar puncture, sputum culture and Gram staining, wound culture, echocardiography, arterial blood gas (ABG) analysis, complete blood count (CBC), basic biochemistry (BMP), lactate concentration, coagulation profile, urinalysis, access to electronic medical records (EMR), execution of EMR-based sepsis algorithms, central venous pressure (CVP) monitoring, and cultures from body parts. The method of claim 10, wherein generating the output indicating that the subject from which the sample was taken is suspected to have an infectious disease comprises generating the output indicating that the subject from which the sample was taken is suspected to have an infectious disease based on the immunophenotyping test and the results of one or more diagnostic tools selected from the group.

14. The method of claim 10, further comprising performing a hemocytometry of the blood sample, and generating the output indicating that the subject from whom the sample was taken is suspected to have an infectious disease, wherein the output indicating that the subject from whom the sample was taken is suspected to have an infectious disease is generated based on the immunophenotyping test and the hemocytometry.

15. The method of claim 10, further comprising identifying the morphological characteristics of entities in the blood sample, and generating the output indicating that the subject from whom the sample was taken is suspected to have an infectious disease, wherein the output indicating that the subject from whom the sample was taken is suspected to have an infectious disease is generated based on the immunophenotyping test and the morphological characteristics of entities in the blood sample.

16. The method according to claim 15, wherein identifying the morphological characteristics of the entity in the blood sample includes identifying at least one of nuclear shape, nuclear density, cytoplasm, and / or cell contour.

17. The method according to claim 10, wherein performing an immunophenotypic test on the blood sample includes determining the average fluorescence intensity level of the fluorescently labeled antibody in one or more fluorescence microscope images of the blood sample.

18. The method according to claim 10, wherein performing the immunophenotypic test includes determining the number of neutrophils expressing CD64 in the blood sample, and generating the output indicating that the subject from whom the blood sample was collected is suspected of having an infectious disease includes generating the output according to the number of neutrophils expressing CD64.

19. The method according to claim 10, wherein performing the immunophenotypic test includes determining the number of lymphocytes expressing CD64 in the blood sample, and generating the output indicating that the subject from whom the blood sample was collected is suspected of having an infectious disease includes generating the output according to the number of lymphocytes expressing CD64.

20. The method according to claim 10, wherein performing the immunophenotypic test includes determining the number of monocytes expressing CD64 in the blood sample, and generating the output indicating that the subject from whom the blood sample was collected is suspected of having an infectious disease includes generating the output according to the number of monocytes expressing CD64.

21. The method according to claim 10, wherein performing the immunophenotypic test includes determining the number of white blood cells expressing CD64 in the blood sample, and generating the output indicating that the subject from whom the blood sample was collected is suspected of having an infectious disease includes generating the output according to the number of white blood cells expressing CD64.

22. The method according to claim 10, wherein performing the immunophenotypic test includes quantifying the expression of CD64 in neutrophils in the blood sample, and generating the output indicating that the subject from whom the blood sample was collected is suspected of having an infectious disease includes generating the output in accordance with the expression of CD64 in the neutrophils in the blood sample.

23. The method according to claim 10, wherein performing the immunophenotypic test includes quantifying the expression of CD64 in lymphocytes in the blood sample, and generating the output indicating that the subject from whom the blood sample was collected is suspected of having an infectious disease includes generating the output in accordance with the expression of CD64 in lymphocytes in the blood sample.

24. The method according to claim 10, wherein performing the immunophenotypic test includes quantifying the expression of CD64 in monocytes in the blood sample, and generating the output indicating that the subject from whom the blood sample was collected is suspected of having an infectious disease includes generating the output in accordance with the expression of CD64 in the monocytes in the blood sample.

25. The method according to claim 10, wherein performing the immunophenotypic test includes quantifying the expression of CD64 in the white blood cells in the blood sample, and generating the output indicating that the subject from whom the blood sample was collected is suspected of having an infectious disease includes generating the output in accordance with the expression of CD64 in the white blood cells in the blood sample.

26. The method according to any one of claims 1 to 6, wherein mixing the fluorescently labeled antibody with the blood sample comprises mixing with an anti-CD19 fluorescently labeled antibody, and performing an immunophenotypic test of the blood sample by analyzing one or more fluorescence microscope images of the blood sample comprises counting B cell lymphocytes in the blood sample.

27. The method according to claim 26, wherein mixing the fluorescently labeled antibody with the blood sample further comprises mixing an anti-CD3 fluorescently labeled antibody, and performing an immunophenotypic test of the blood sample by analyzing one or more fluorescence microscope images of the blood sample comprises counting T-cell lymphocytes in the blood sample.

28. The method according to claim 27, wherein generating the output includes generating an output indicating the relative number of B-cell lymphocytes and T-cell lymphocytes in the blood sample.

29. The method according to any one of claims 1 to 9, further comprising: identifying morphological features of entities within a blood sample by analyzing one or more microscopic images of the blood sample; determining the characteristics of the blood sample based on the morphological features of entities within the sample and the immunophenotypic test, wherein generating an output includes generating an output in accordance with the determination of the characteristics of the blood sample.

30. The method according to claim 29, wherein identifying the morphological features of the entity in the sample includes identifying at least one of nuclear shape, nuclear density, cytoplasm, and / or cell contour.

31. A device for use with a sample chamber that contains a blood sample mixed with a fluorescently labeled antibody, An optical measurement unit configured to receive the sample chamber, comprising a microscope configured to acquire one or more fluorescence microscope images of the blood sample in the sample chamber, At least one computer processor, By analyzing one or more fluorescence microscope images of the blood sample, an immunophenotypic test is performed on the blood sample. A computer processor configured to generate an output in accordance with this, at least partially, A device equipped with the following features.

32. The aforementioned computer processor further, By analyzing one or more microscopic images of the blood sample, the number of entities in the blood sample is counted. The system is configured to determine the characteristics of the blood sample based on the count of entities in the sample and the immunophenotypic test. The apparatus according to claim 31.

33. Apparatus for use with a sample chamber containing a blood sample mixed with a fluorescently labeled antibody against one or more cell surface markers listed in Table 1, wherein at least one computer processor is configured to perform an immunophenotypic test on the blood sample by identifying an index corresponding to the one or more cell surface markers listed in Table 1.

34. The apparatus according to claim 31, wherein the computer processor is configured to perform an immunophenotypic test of a blood sample by analyzing one or more fluorescence microscopy images of the blood sample, by determining the average fluorescence intensity level of cells bound to the fluorescently labeled antibody in one or more of the microscope images of the blood sample.

35. The apparatus according to claim 31, for use in conjunction with a sample chamber containing a blood sample mixed with an anti-CD3 fluorescently labeled antibody, wherein the computer processor is configured to perform the immunophenotyping test by identifying T cell lymphocytes in the blood sample.

36. The apparatus according to claim 31, for use in conjunction with a sample chamber containing the blood sample mixed with an anti-CD3 fluorescently labeled antibody, wherein the computer processor is configured to perform the immunophenotypic test by counting T cell lymphocytes in the blood sample.

37. The apparatus for use with a sample chamber containing a blood sample mixed with an anti-CD19 fluorescently labeled antibody, wherein the computer processor is configured to perform the immunophenotyping test by identifying B cell lymphocytes in the blood sample.

38. Apparatus for use with a sample chamber containing the blood sample mixed with an anti-CD64 fluorescently labeled antibody, wherein the computer processor is configured to generate an output that, at least in part, the subject from whom the blood sample was taken is suspected to be infected with an infectious disease, according to any one of claims 31 or 33 to 37.

39. The apparatus according to claim 38, wherein the computer processor is configured to generate an output indicating that the subject from whom the blood sample was taken is suspected to have sepsis, at least in part based on the immunophenotypic test.

40. The aforementioned computer processor Identify one or more biomarkers selected from the group consisting of procalcitonin (PCT), C-reactive protein (CRP), interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-10 (IL-10), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), presepsin, thrombomodulin, lactate-soluble urokinase-type plasminogen-activating factor receptor (suPAR), soluble trigger receptor-1 (sTREM-1) expressed in bone marrow cells, lipopolysaccharide-binding protein (LBP), N-terminal pro-brain natriuretic peptide (NT-proBNP), neutrophil gelatinase-binding lipocalin (NGAL), adrenomedullin, resistin, pro-adrenomedullin (MR-proADM), D-dimer, and pancreatic stone protein (PSP). Based on the immunophenotypic test and one or more biomarkers selected from the group, the system is configured to generate an output indicating that the subject from whom the sample was taken is suspected of having the infectious disease. The apparatus according to claim 38.

41. The aforementioned computer processor The results of one or more diagnostic tests selected from the group consisting of vital sign measurement, blood culture, urine culture, chest X-ray, CT scan, ultrasound, lumbar puncture, sputum culture and Gram staining, wound culture, echocardiography, arterial blood gas (ABG) analysis, complete blood count (CBC), basic biochemistry (BMP), lactate concentration, coagulation profile, urinalysis, access to electronic medical records (EMR), execution of EMR-based sepsis algorithms, central venous pressure (CVP) monitoring, and cultures from body parts are received. Based on the immunophenotypic test and the results of one or more diagnostic tools selected from the group, the system is configured to generate an output indicating that the subject from whom the sample was taken is suspected to be suffering from the infectious disease. The apparatus according to claim 38.

42. The apparatus according to claim 38, wherein the computer processor is configured to perform a blood cell count of the blood sample and to generate an output indicating that the subject from whom the sample was taken is suspected to be suffering from the infectious disease, based on the immunophenotyping test and the blood cell count.

43. The apparatus according to claim 38, wherein the computer processor is configured to identify the morphological characteristics of entities in the blood sample and to generate an output indicating that the subject from whom the sample was taken is suspected of having the infectious disease, based on the immunophenotyping test and the morphological characteristics of entities in the blood sample.

44. The method according to claim 43, wherein the computer processor is configured to identify the morphological characteristics of entities in the blood sample by identifying at least one of nuclear shape, nuclear density, cytoplasm, and / or cell contour.

45. The apparatus according to claim 38, wherein the computer processor is configured to perform an immunophenotypic test of a blood sample by analyzing one or more fluorescence microscopy images of the blood sample, by determining the average fluorescence intensity level of cells bound to the CD64 fluorescently labeled antibody in one or more of the microscope images of the blood sample.

46. The apparatus according to claim 38, wherein the computer processor is configured to determine the number of neutrophils expressing CD64 in the blood sample and to generate the output based on the number of neutrophils expressing CD64.

47. The apparatus according to claim 38, wherein the computer processor is configured to determine the number of lymphocytes expressing CD64 in the blood sample and to generate the output based on the number of lymphocytes expressing CD64.

48. The apparatus according to claim 38, wherein the computer processor is configured to determine the number of monocytes expressing CD64 in the blood sample and to generate the output based on the number of monocytes expressing CD64.

49. The apparatus according to claim 38, wherein the computer processor is configured to determine the number of white blood cells expressing CD64 in the blood sample and to generate the output based on the number of white blood cells expressing CD64.

50. The apparatus according to claim 38, wherein the computer processor is configured to quantify the CD64 expression in neutrophils in the blood sample and to generate the output based on the CD64 expression in neutrophils in the blood sample.

51. The apparatus according to claim 38, wherein the computer processor is configured to quantify the CD64 expression in lymphocytes in the blood sample and to generate the output based on the CD64 expression in lymphocytes in the blood sample.

52. The apparatus according to claim 38, wherein the computer processor is configured to quantify the CD64 expression in monocytes in the blood sample and to generate the output based on the CD64 expression in monocytes in the blood sample.

53. The apparatus according to claim 38, wherein the computer processor is configured to determine the CD64 expression in the white blood cells in the blood sample and to generate the output based on the CD64 expression in the white blood cells in the blood sample.

54. The apparatus for use in conjunction with a sample chamber containing a blood sample mixed with an anti-CD19 fluorescently labeled antibody, wherein the computer processor is configured to perform the immunophenotypic test by counting B-cell lymphocytes in the blood sample, according to any one of claims 31 to 34.

55. The apparatus for use with a sample chamber containing a blood sample mixed with an anti-CD3 fluorescently labeled antibody and an anti-CD19 fluorescently labeled antibody, wherein the computer processor is configured to perform the immunophenotypic test by counting T-cell lymphocytes and B-cell lymphocytes in the blood sample.

56. The apparatus according to claim 55, wherein the computer processor is configured to generate the output by generating an output indicating the relative number of B-cell lymphocytes and T-cell lymphocytes in the blood sample.

57. The aforementioned computer processor further, By analyzing one or more microscopic images of the blood sample, the morphological characteristics of the organisms within the sample are identified. The apparatus according to any one of claims 31 to 37, configured to determine the characteristics of the blood sample based on the morphological characteristics of the entity in the sample and the immunophenotypic test.

58. The apparatus according to claim 57, wherein the computer processor is configured to identify the morphological characteristics of entities in the sample by identifying at least one of nuclear shape, nuclear density, cytoplasm, and / or cell contour.

59. Placing a blood sample into the sample chamber, The sample chamber containing the blood sample is placed inside the optical measurement unit, Using the microscope of the optical measurement unit, one or more microscopic images of the blood sample in the sample chamber are acquired. Using a computer processor, The process involves analyzing one or more microscopic images of the blood sample to count the actual substances within the blood sample, By analyzing one or more microscopic images of the blood sample, an immunophenotypic test is performed on the blood sample. Based on the count of entities in the sample and the immunophenotypic test, the characteristics of the blood sample are determined. To generate output in accordance with this, at least partially. A method that includes this.

60. The method according to claim 59, wherein the immunophenotyping test comprises mixing a fluorescently labeled antibody against one or more cell surface markers listed in Table 1 with the sample, and identifying an index corresponding to the one or more cell surface markers listed in Table 1.

61. The method according to claim 59, further comprising mixing a fluorescently labeled antibody with the blood sample.

62. The method according to claim 59, wherein the counting of entities includes counting at least one of red blood cells, neutrophils, eosinophils, basophils, lymphocytes, monocytes, and / or platelets.

63. The method according to claim 59, wherein performing an immunophenotypic test includes performing an immunophenotypic test for CD3, and generating the output includes generating an output indicating the number of T cell lymphocytes in the blood sample.

64. The method according to claim 59, wherein performing an immunophenotypic test includes performing an immunophenotypic test for CD19, and generating the output includes generating an output indicating the number of B-cell lymphocytes in the blood sample.

65. The method according to claim 59, wherein performing an immunophenotypic test comprises performing an immunophenotypic test for CD3 and CD19, and generating the output comprises generating an output indicating the number of B-cell lymphocytes and T-cell lymphocytes in the blood sample.

66. The method according to claim 59, wherein performing an immunophenotyping test includes performing an immunophenotyping test for at least one of CD14 and CD16.

67. The method according to any one of claims 59 to 66, wherein performing an immunophenotypic test includes performing an immunophenotypic test for CD64, and generating the output includes generating an output indicating that the subject from which the sample was taken is suspected to have an infectious disease.

68. The apparatus according to claim 67, wherein generating the output includes generating an output indicating that the subject from whom the sample was taken is suspected to be suffering from sepsis.

69. A device for use with a sample chamber for containing blood samples, An optical measuring unit configured to receive the sample chamber, comprising a microscope configured to acquire one or more microscopic images of the blood sample in the sample chamber, At least one computer processor, By analyzing one or more microscopic images of the blood sample, the number of entities in the blood sample is counted. By analyzing one or more microscopic images of the blood sample, an immunophenotypic test is performed on the blood sample. Based on the count of entities in the sample and the immunophenotypic test, the characteristics of the blood sample are determined. A computer processor configured to generate an output in accordance with this, at least partially, A device equipped with the following features.

70. Apparatus for use with a sample chamber containing a blood sample mixed with a fluorescently labeled antibody against one or more cell surface markers listed in Table 1, wherein at least one computer processor is configured to perform an immunophenotypic test of the blood sample by identifying an index corresponding to the one or more cell surface markers listed in Table 1.

71. The apparatus according to claim 69, wherein the computer processor is configured to perform a count of entities by counting at least one of red blood cells, neutrophils, eosinophils, basophils, lymphocytes, monocytes, and / or platelets.

72. The apparatus according to claim 69, wherein the computer processor is configured to perform an immunophenotypic test by performing an immunophenotypic test for CD3, and the computer processor is configured to generate an output indicating the number of T cells in the blood sample.

73. The apparatus according to claim 69, wherein the computer processor is configured to perform an immunophenotypic test by performing an immunophenotypic test for CD19, and the computer processor is configured to generate an output indicating the number of B cell lymphocytes in the blood sample.

74. The apparatus according to claim 69, wherein the computer processor is configured to perform an immunophenotypic test by performing an immunophenotypic test on CD3 and CD19, and the computer processor is configured to generate an output indicating the number of B-cell lymphocytes and T-cell lymphocytes in the blood sample.

75. The apparatus according to claim 69, wherein the computer processor is configured to perform an immunophenotypic test by performing an immunophenotypic test on at least one of CD14 and CD16.

76. The apparatus according to any one of claims 69 to 75, wherein the computer processor is configured to perform an immunophenotypic test by performing an immunophenotypic test on CD64, and the computer processor is configured to generate an output indicating that the subject from whom the sample was taken is suspected to be suffering from an infectious disease.

77. The apparatus according to claim 76, wherein the computer processor is configured to generate the output by generating an output indicating that the subject from whom the sample was taken is suspected to be suffering from sepsis.

78. Placing a blood sample into the sample chamber, The sample chamber containing the blood sample is placed inside the optical measurement unit, Using the microscope of the optical measurement unit, one or more microscopic images of the blood sample in the sample chamber are acquired. Using a computer processor, By analyzing one or more microscopic images of the blood sample, the morphological characteristics of the organisms within the sample are identified. By analyzing one or more microscopic images of the blood sample, an immunophenotypic test is performed on the blood sample. The characteristics of the blood sample are determined based on the morphological characteristics of the entity in the sample and the immunophenotypic test, To generate output in accordance with this, at least partially. A method that includes this.

79. The method according to claim 78, wherein the immunophenotyping test comprises mixing a fluorescently labeled antibody against one or more cell surface markers listed in Table 1 with the sample, and identifying an index corresponding to the one or more cell surface markers listed in Table 1.

80. The method according to claim 78, further comprising mixing a fluorescently labeled antibody with the blood sample.

81. The method according to claim 78, wherein identifying the morphological features of the entities in the sample includes identifying at least one of nuclear shape, nuclear density, cytoplasmic shape, cytoplasmic granularity, and / or cell contour.

82. The method according to claim 78, wherein performing an immunophenotypic test includes performing an immunophenotypic test for CD3, and generating the output includes generating an output indicating the number of T cell lymphocytes in the blood sample.

83. The method according to claim 78, wherein performing an immunophenotypic test includes performing an immunophenotypic test for CD19, and generating the output includes generating an output indicating the number of B-cell lymphocytes in the blood sample.

84. The method according to claim 78, wherein performing an immunophenotypic test comprises performing an immunophenotypic test for CD3 and CD19, and generating the output comprises generating an output indicating the number of B-cell lymphocytes and T-cell lymphocytes in the blood sample.

85. The method according to claim 78, wherein performing an immunophenotyping test includes performing an immunophenotyping test for at least one of CD14 and CD16.

86. The method according to any one of claims 78 to 85, wherein performing an immunophenotypic test includes performing an immunophenotypic test for CD64, and generating the output includes generating an output indicating that the subject from which the sample was taken is suspected to have an infectious disease.

87. The apparatus according to claim 86, wherein generating the output includes generating an output indicating that the subject from whom the sample was taken is suspected to be suffering from sepsis.

88. A device for use with a sample chamber for containing blood samples, An optical measuring unit configured to receive the sample chamber, comprising a microscope configured to acquire one or more microscopic images of the blood sample in the sample chamber, At least one computer processor, By analyzing one or more microscopic images of the blood sample, the morphological characteristics of the organisms within the sample are identified. By analyzing one or more microscopic images of the blood sample, an immunophenotypic test is performed on the blood sample. Based on the morphological characteristics of the substance in the sample and the immunophenotypic test, the characteristics of the blood sample are determined. A computer processor configured to generate an output in accordance with this, at least partially, A device equipped with the following features.

89. Apparatus for use with a sample chamber containing a blood sample mixed with a fluorescently labeled antibody against one or more cell surface markers listed in Table 1, wherein at least one computer processor is configured to perform an immunophenotypic test of the blood sample by identifying an index corresponding to the one or more cell surface markers listed in Table 1.

90. The apparatus according to claim 88, wherein the computer processor is configured to identify the morphological characteristics of entities in the sample by identifying at least one of nuclear shape, nuclear density, cytoplasmic shape, cytoplasmic granularity, and / or cell contour.

91. The apparatus according to claim 88, wherein the computer processor is configured to perform an immunophenotypic test by performing an immunophenotypic test for CD3, and the computer processor is configured to generate an output indicating the number of T cells in the blood sample.

92. The apparatus according to claim 88, wherein the computer processor is configured to perform an immunophenotypic test by performing an immunophenotypic test for CD19, and the computer processor is configured to generate an output indicating the number of B-cell lymphocytes in the blood sample.

93. The apparatus according to claim 88, wherein the computer processor is configured to perform an immunophenotypic test by performing an immunophenotypic test on CD3 and CD19, and the computer processor is configured to generate an output indicating the number of B-cell lymphocytes and T-cell lymphocytes in the blood sample.

94. The apparatus according to claim 88, wherein the computer processor is configured to perform an immunophenotypic test by performing an immunophenotypic test on at least one of CD14 and CD16.

95. The apparatus according to any one of claims 88 to 94, wherein the computer processor is configured to perform an immunophenotypic test by performing an immunophenotypic test on CD64, and the computer processor is configured to generate an output indicating that the subject from which the sample was taken is suspected to be suffering from an infectious disease.

96. The apparatus according to claim 95, wherein the computer processor is configured to generate the output by generating an output indicating that the subject from whom the sample was taken is suspected to be suffering from sepsis.

97. During the training phase, the blood sample analysis system was used. Mixing antibodies fluorescently labeled with fluorescent dyes with multiple blood samples, The fluorescent imaging modality configured to excite the aforementioned fluorescent dye is used to acquire one or more fluorescence microscope images of each blood sample. The second imaging modality involves acquiring one or more separate microscopic images of each blood sample, Using a computer processor, By analyzing one or more fluorescence microscope images of each blood sample, an immunophenotypic test is performed on each blood sample. By analyzing one or more separate microscopic images of each blood sample, the characteristics of each blood sample can be identified. To identify the correlation between the results of the immunophenotypic test and the characteristics of the blood sample that can be identified in the microscopic image acquired by the second imaging modality, A method that includes training to perform immunophenotypic testing of test blood samples without using antibodies.

98. The method according to claim 97, wherein mixing the antibody fluorescently labeled with the fluorescent dye with the plurality of blood samples comprises mixing a fluorescently labeled antibody against one or more cell surface markers listed in Table 1 with the plurality of blood samples, and performing an immunophenotypic test of the blood samples by analyzing one or more fluorescence microscope images of each blood sample comprises identifying an index corresponding to one or more cell surface markers listed in Table 1 in at least some of the blood samples.

99. The method according to claim 97, further comprising, in the next step, performing an immunophenotypic test on the test blood sample without using an antibody.

100. Mixing the antibody fluorescently labeled with the fluorescent dye with the plurality of blood samples includes mixing the anti-CD64 antibody fluorescently labeled with the fluorescent dye with the plurality of blood samples. Identifying the characteristics of each blood sample includes identifying the morphological characteristics of the entities within each blood sample. The method according to claim 99, further comprising determining that the subject from whom the test blood sample was collected is suspected to be suffering from an infectious disease, in accordance with an immunophenotypic test of the test blood sample without the use of antibodies.

101. The method according to claim 100, wherein identifying the morphological characteristics of entities in each blood sample includes identifying at least one of the nuclear shape, nuclear density, cytoplasmic shape, cytoplasmic granularity, and / or cell contour of entities in each blood sample.

102. A blood analysis system equipped with a microscope, At least one computer processor associated with the blood analysis system, wherein during the training phase, To receive one or more fluorescence microscope images of each of several blood samples mixed with an antibody fluorescently labeled with a fluorescent dye, the fluorescence microscope images being acquired with a fluorescence imaging modality configured to excite the fluorescent dye, To receive one or more separate microscopic images of each blood sample, and one or more separate microscopic images acquired with a second imaging modality, By analyzing one or more fluorescence microscope images of each blood sample, an immunophenotypic test is performed on each blood sample. By analyzing one or more separate microscopic images of each blood sample, the characteristics of each blood sample can be identified. To identify the correlation between the results of the immunophenotypic test and the characteristics of the blood sample that can be identified in the microscopic image acquired by the second imaging modality, A system comprising at least one computer processor configured to train the blood sample analysis system to perform immunophenotypic testing of test blood samples without using antibodies, A device equipped with the following features.

103. The apparatus for use with a sample chamber containing a blood sample mixed with a fluorescently labeled antibody against one or more cell surface markers listed in Table 1, wherein the at least one computer processor is configured to perform an immunophenotypic test on the blood sample by identifying an index corresponding to the one or more cell surface markers listed in Table 1.

104. The apparatus according to claim 102, wherein the computer processor is configured to perform an immunophenotypic test of the test blood sample without using an antibody in the next step.

105. An apparatus for use with a sample chamber that contains a blood sample in which a fluorescently labeled anti-CD64 antibody, which has been fluorescently labeled with a fluorescent dye, has been mixed with multiple blood samples, The aforementioned computer processor By identifying the morphological characteristics of the organisms within each blood sample, the characteristics of each blood sample can be identified. The system is configured to determine whether the subject from whom the test blood sample was collected is suspected of having an infectious disease, based on the immunophenotypic testing of the test blood sample without the use of antibodies. The apparatus according to claim 104.

106. The apparatus according to claim 105, wherein the computer processor is configured to identify the morphological characteristics of entities in each blood sample by identifying at least one of the nuclear shape, nuclear density, cytoplasmic shape, cytoplasmic granularity, and / or cell contour of entities in each blood sample.

107. A method for use in a training phase with a computer processor trained to identify features that correlate with antibody expression in a training blood sample, Obtain at least one microscopic image of a test blood sample that does not contain added antibodies, Using a computer processor, By analyzing the aforementioned microscopic images and identifying features that are determined to correlate with antibody expression in the training blood sample during the training phase, an immunophenotypic test of the test blood sample is performed. To generate output accordingly, A method that includes this.

108. A computer processor trained to identify features that correlate with antibody expression in a training blood sample during the training phase, We receive one or more microscopic images of test blood samples that do not contain added antibodies. By analyzing the aforementioned microscopic images and identifying features that are determined to correlate with antibody expression in the training blood sample during the training phase, an immunophenotypic test is performed on the test blood sample. A computer processor configured to generate output accordingly. A device equipped with the following features.

109. A method for use with blood samples, Obtain one or more microscopic images of the aforementioned blood sample, Using a computer processor, By analyzing one or more of the aforementioned microscope images, neutrophils expressing CD64 in the blood sample are identified, To generate output accordingly, A method that includes this.

110. A method for use with a blood sample that has not had an antibody added, comprising identifying neutrophils that express CD64 in the blood sample, wherein identifying neutrophils that express CD64 in the blood sample that has not had an antibody added.

111. A method for use with a blood sample to which an anti-CD64 antibody fluorescently labeled with a fluorescent dye has been added, wherein identifying neutrophils expressing CD64 in the blood sample includes identifying the anti-CD64 antibody fluorescently labeled with the fluorescent dye in the microscopic image.

112. The method according to any one of claims 109 to 111, further comprising determining the number of neutrophils expressing CD64 in the blood sample.

113. The method according to claim 112, wherein generating the output includes outputting the number of neutrophils expressing CD64 in the blood sample.

114. The method according to any one of claims 109 to 111, further comprising determining the percentage of neutrophils expressing CD64 in the blood sample.

115. The method according to claim 8b, wherein generating the output includes outputting the percentage of neutrophils expressing CD64 in the blood sample.

116. The method according to any one of claims 109 to 111, further comprising using the computer processor to determine the quantification of CD64 expression in the neutrophils in the blood sample.

117. The method according to 116, wherein generating the output includes generating an output indicating that the subject from whom the sample was taken is suspected to have an infectious disease, based on the quantification of CD64 expression in the neutrophils in the blood sample.

118. The method further includes identifying one or more biomarkers selected from the group consisting of procalcitonin (PCT), C-reactive protein (CRP), interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-10 (IL-10), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), presepsin, thrombomodulin, lactate-soluble urokinase-type plasminogen-activating factor receptor (suPAR), soluble trigger receptor-1 (sTREM-1) expressed in bone marrow cells, lipopolysaccharide-binding protein (LBP), N-terminal pro-brain natriuretic peptide (NT-proBNP), neutrophil gelatinase-binding lipocalin (NGAL), adrenomedullin, resistin, pro-adrenomedullin (MR-proADM), D-dimer, and pancreatic stone protein (PSP). The method according to claim 117, wherein generating the output indicating that the subject from whom the sample was collected is suspected to be suffering from an infectious disease comprises generating the output indicating that the subject from whom the sample was collected is suspected to be suffering from an infectious disease based on the quantification of CD64 expression in the neutrophils in the blood sample and one or more biomarkers selected from the group.

119. This further includes receiving the results of one or more diagnostic tests selected from the group consisting of vital sign measurement, blood culture, urine culture, chest X-ray, CT scan, ultrasound, lumbar puncture, sputum culture and Gram staining, wound culture, echocardiography, arterial blood gas (ABG) analysis, complete blood count (CBC), basic biochemistry (BMP), lactate concentration, coagulation profile, urinalysis, access to electronic medical records (EMR), execution of EMR-based sepsis algorithms, central venous pressure (CVP) monitoring, and cultures from body parts. The method according to claim 117, wherein generating the output indicating that the subject from whom the sample was taken is suspected to be suffering from an infectious disease includes generating the output indicating that the subject from whom the sample was taken is suspected to be suffering from an infectious disease based on the quantification of CD64 expression in the neutrophils in the blood sample and the results of one or more diagnostic tools selected from the group.

120. The method according to claim 117, further comprising performing a hemocytometry of the blood sample, and generating the output indicating that the subject from whom the sample was taken is suspected to have an infectious disease, wherein the output is generated based on the quantification of CD64 expression in the neutrophils in the blood sample and the hemocytometry, indicating that the subject from whom the sample was taken is suspected to have an infectious disease.

121. The method according to claim 117, further comprising identifying the morphological characteristics of entities in the blood sample, and generating the output indicating that the subject from whom the sample was taken is suspected of having an infectious disease, wherein the output indicating that the subject from whom the sample was taken is suspected of having an infectious disease is generated based on the quantification of CD64 expression in the neutrophils in the blood sample and the morphological characteristics of entities in the blood sample.

122. The method according to claim 121, wherein identifying the morphological characteristics of the entity in the blood sample includes identifying at least one of nuclear shape, nuclear density, cytoplasm, and / or cell contour.

123. Placing a blood sample into the sample chamber, The sample chamber containing the blood sample is placed inside the optical measurement unit, Using the microscope of the optical measurement unit, one or more microscopic images of the blood sample in the sample chamber are acquired. Using a computer processor, By analyzing one or more microscopic images of the blood sample, the morphological characteristics of the organisms within the sample are identified. Based on the identification of the aforementioned morphological characteristics, it is determined that the subject from whom the sample was taken is suspected of having an infectious disease, To generate output in accordance with this, at least partially. A method that includes this.

124. The method according to claim 123, further comprising using the computer processor to identify one or more cell surface markers listed in Table 1, wherein determining that the subject from which the sample was taken is suspected of having an infectious disease is based on the identification of the morphological characteristics and the identification of one or more cell surface markers listed in Table 1.

125. The method according to claim 123 or claim 124, further comprising using the computer processor to identify one or more cell markers listed in Table 2, wherein determining that the subject from which the sample was taken is suspected of having an infectious disease is based on the identification of the morphological characteristics and the identification of one or more cell markers listed in Table 2.

126. The method according to claim 125, further comprising using the computer processor to identify the expression level of CD64 in the blood sample, wherein determining that the subject from whom the sample was collected is suspected of having an infectious disease is based on the identification of the morphological characteristics and the identification of the expression level of CD64 in the sample.

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