Optical measurements performed on the sample

Abstract

Apparatus and methods for a blood sample (48, 50) are described that include measuring a hemoglobin concentration of at least a portion (48) of the blood sample by performing a first measurement on the blood sample. The mean corpuscular hemoglobin of the blood sample (48, 50) is measured by performing a second measurement on the blood sample (48, 50). A parameter of the blood sample is determined based on a relationship between the hemoglobin concentration and the mean corpuscular hemoglobin. Other applications are described.

Description

[0001] This application is a continuation-in-part of the application entitled "Optical Measurements Performed on a Sample" having application number 2017800279083, filing date May 11, 2017.

[0002] Cross Reference to Related Applications

[0003] This application claims priority to U.S. Provisional Patent Application No. 62 / 334,517, entitled "Method and Apparatus for Estimating Dilution and Concentration," by Zait, filed May 11, 2016.

[0004] This application is related to the International Application entitled "Sample carrier for optical measurements," filed on the same day as the present application, which claims priority to U.S. Provisional Patent Application entitled "Sample carrier for optical measurements," by Pollak, filed May 11, 2016.

[0005] The above-referenced applications are incorporated herein by reference. TECHNICAL FIELD

[0006] Some applications of the presently disclosed subject matter relate generally to analyzing biological samples, particularly by performing optical measurements. BACKGROUND

[0007] There are a variety of methods for quantifying parameters in a sample, such as a blood sample. In some such methods, the sample is diluted and then analyzed. For example, a blood sample can be diluted in order to increase the visibility of sample components in a microscopic image of the sample, and / or a staining substance can be added to the blood sample, thereby staining given components in the sample.

[0008] In some cases, a sample is analyzed using more than one type of measurement device. For example, a microscope is sometimes used in order to analyze individual cells in a sample, while an imaging device such as a spectrographic camera is used to analyze the sample at a bulk level (e.g., by performing optical absorption, transmission, fluorescence, and / or luminescence measurements). SUMMARY

[0009] According to some applications of the present application, a portion of the blood sample is diluted using dilution techniques, such as the techniques described by Pollak in US 2015 / 0316477, which is incorporated herein by reference. Typically, the portion of the blood sample is imaged using a microscope system, which can be manual or automated. For some applications, the microscopic images are analyzed (e.g., manually or using a computer processor running suitable computer software) to identify different blood cells.

[0010] For some applications of the present application, variations and / or errors that occur during dilution have been accounted for. Typically, a 10% error in the dilution factor occurs, which can directly correspond to a 10% error in, for example, the count of red blood cells per unit volume (e.g., per microliter) of blood. Such dilution errors can arise from a variety of sources. Illustrative examples of such error sources, which are not intended to limit the scope of the present application, include inaccuracy or error of a pipette, inaccuracy or error of calibration, inaccuracy or error of mixing, etc. Accordingly, according to some applications of the present application, measurements are made on a source sample portion (e.g., an undiluted blood sample portion), from which the diluted sample portion is extracted. Such measurements typically correspond to at least one measurement made on the diluted sample portion. For example, measurements made on the source sample portion can include measurements of: hemoglobin content, white blood cell content, red blood cell content, hematocrit, content of specific white blood cell types, platelet content, and / or any measurement that is measured or can be inferred for the diluted sample portion. For some applications, a normalization factor is determined, which is a property of the source sample portion that is related to other measurements (e.g., the number of red blood cells per unit area or the number of red blood cells per unit volume in the source sample portion). Typically, based on the normalization factor, the measured variable in the sample (e.g., the source sample portion) is determined, as described in more detail below.

[0011] For some applications, hematocrit is measured by performing a first measurement on a blood sample, and mean corpuscular volume is measured by performing a second measurement on the blood sample. For example, hematocrit can be measured using a microhematocrit method (in which blood is centrifuged), and / or by performing ultrasound and / or impedance measurements on a first portion of a blood sample, and mean corpuscular volume can be measured by analyzing microscopic images obtained from a second portion of the blood sample. Typically, the second portion of the sample is diluted relative to the first portion of the blood sample, for example to improve visibility of individual cells, to stain the second portion of the sample, and / or for a different reason. For some applications, a relationship between the first portion of the sample and the second portion of the sample is determined based on a relationship between hematocrit and mean corpuscular volume. For some applications, a parameter of the source sample portion is determined based on a relationship between hematocrit and mean corpuscular volume. Typically, red blood cell count in the sample (e.g., count per unit volume) is determined by dividing hematocrit by mean corpuscular volume. For some applications, a count of one or more other components in the sample (e.g., red blood cells of a given type, white blood cells, white blood cells of a given type, circulating tumor cells, platelets, platelets of a given type, bacteria, pathogens, pathogens of a given type, reticulocytes, and / or Howell-Jolly bodies) is determined based on the red blood cell count in the sample. For example, a ratio of red blood cell count to count of one or more other components in the sample portion is determined by analyzing microscopic images of the sample portion. Then, a count of one or more other components can be determined based on the red blood cell count in the sample, and the ratio of red blood cell count to count of one or more other components in the sample portion.

[0012] For some applications, hemoglobin concentration is measured by performing a first measurement on a blood sample, and mean corpuscular hemoglobin in the blood sample is measured by performing a second measurement on the blood sample. For example, hemoglobin concentration can be measured by performing an optical density measurement on a first portion of the blood sample, and mean corpuscular hemoglobin is measured by analyzing fibergram images obtained from a second portion of the blood sample. Typically, the second portion of the sample is diluted relative to the first portion of the blood sample, for example to improve visibility of individual cells, and / or to stain the second portion of the sample, and / or for a different reason. For some applications, a relationship between the first portion of the sample and the second portion of the sample is determined based on a relationship between hemoglobin concentration and mean corpuscular hemoglobin. For some applications, a parameter of the source sample portion is determined based on a relationship between hemoglobin concentration and mean corpuscular hemoglobin. Typically, red blood cell count in the sample (e.g., count per unit volume) is determined by dividing hemoglobin concentration by mean corpuscular hemoglobin. For some applications, a count of one or more other components in the sample (e.g., red blood cells of a given type, white blood cells, white blood cells of a given type, circulating tumor cells, platelets, platelets of a given type, bacteria, pathogens, pathogens of a given type, reticulocytes, and / or Howell-Jolly bodies) is determined based on red blood cell count in the sample. The ratio of red blood cell count to count of one or more other components in the sample portion is determined, for example by analyzing microscopic images of the sample portion. Then, a count of one or more other components can be determined based on red blood cell count in the sample, and the ratio of red blood cell count to count of one or more other components in the sample portion.

[0013] For some applications of the application, two or more measurements are made on a biological sample, which are typically optical measurements. Typically, the biological sample is a blood sample. For some applications, a bulk level of a measured variable of the sample is measured by a first measurement on the sample, and a cell level of the measured variable of the sample is measured by a second measurement on the sample. For the purposes of this application, the term "cell level of a measured variable" is understood to mean a measured variable relating to one or more parameters of individual cells or other non-dissolved constituents in the sample, such as mean corpuscular volume, mean corpuscular hemoglobin, mean platelet volume, and / or the like. Measurement of a cell level of a measured variable typically involves a first step of identifying individual cells or other non-dissolved constituents in the sample (such as identifying such constituents in a fibergram image), and a second step of identifying parameters of such individually identified constituents. Typically, a cell level of a measured variable is measured by analyzing one or more microscopic images of the sample. For the purposes of this application, the term "bulk level of a measured variable" is understood to mean a measured variable relating to a parameter of the sample as a whole, and which does not require the two steps of identifying individual cells or other non-dissolved constituents in the sample and identifying parameters of such individually identified constituents. For example, such measured variables can include optical density of a given constituent (which is measured by a measurement on a bulk volume of the sample, such as even after dissolution of individual constituents in the bulk volume), count / unit volume of a given constituent (which is typically measured by identifying such constituents, but without identifying parameters of individual identified constituents), and / or concentration of a given constituent (such as red blood cell concentration, hemoglobin concentration, white blood cell concentration, platelet concentration, and / or hematocrit). Typically, a bulk level of a measured variable is measured by a measurement on a bulk volume of the sample. For example, such measurements can include ultrasonic, impedance, optical absorption, transmission, fluorescence, microscopic, and / or luminescence measurements on a bulk volume of the sample. Typically, a parameter of the sample is determined based on a relationship between the bulk level of the measured variable and the cell level of the measured variable.

[0014] For some applications, the first and second optical measurements on the sample are made using one or more optical measurement devices under different sets of measurement conditions from each other. A measured variable of the sample is measured based on the first optical measurement, and a measured variable of the sample is measured based on the second optical measurement. Depending on the application, the measured variable measured based on the second optical measurement is the same as the measured variable measured based on the first optical measurement, or is different from the measured variable measured based on the first optical measurement. Depending on the application, the first and second optical measurements are made on the same portion of the sample, or on different portions of the sample. For some applications, one optical measurement is made on a portion of the sample, wherein the portion of the sample is diluted relative to the portion of the sample on which the other optical measurement is made.

[0015] Typically, a relationship between measurement conditions of one or more optical measurement devices used to perform the first and second optical measurements is determined based on a relationship between the measured variable measured on the basis of the first optical measurement and the measured variable measured on the basis of the second optical measurement. For example, the first and second optical measurements can be performed on respective portions of a sample, wherein the respective portions of the sample are disposed in respective portions of one or more sample chambers having respective dimensions (e.g., respective heights). For some such applications, a relationship between the dimensions of the respective portions of the one or more sample chambers is determined based on a relationship between the measured variable measured on the basis of the first optical measurement and the measured variable measured on the basis of the second optical measurement. Alternatively or additionally, a field of view range is determined based on a relationship between the measured variable measured on the basis of the first optical measurement and the measured variable measured on the basis of the second optical measurement, wherein one of the first and second optical measurements (e.g., a micrograph) is measured by the field of view range, and / or a level of magnification is determined based on a relationship between the measured variable measured on the basis of the first optical measurement and the measured variable measured on the basis of the second optical measurement, wherein one of the first and second optical measurements (e.g., a micrograph) is measured at the level of magnification. For some applications, the first and second measurements are normalized with respect to one another. Subsequently, a parameter of the sample is determined based on the normalization of the first and second measurements with respect to one another.

[0016] According to some applications of the present application, therefore, there is provided a method for a blood sample, the method comprising:

[0017] - measuring a hemoglobin concentration in at least a portion of the blood sample by performing a first measurement on the blood sample;

[0018] - measuring a mean corpuscular hemoglobin of the blood sample by performing a second measurement on the blood sample; and

[0019] - determining a parameter of the blood sample based on a relationship between the hemoglobin concentration and the mean corpuscular hemoglobin.

[0020] In some applications, determining the parameter of the blood sample comprises normalizing the first and second measurements with respect to one another based on the relationship between the hemoglobin concentration and the mean corpuscular hemoglobin.

[0021] In some applications, performing the first measurement on the blood sample comprises performing an optical density measurement on the blood sample.

[0022] In some applications, measuring the hemoglobin concentration in at least a portion of the blood sample comprises measuring the hemoglobin concentration in a first portion of the blood sample, measuring the mean corpuscular hemoglobin comprises measuring the mean corpuscular hemoglobin in a second portion of the blood sample, and determining the parameter of the sample comprises determining a relationship between the first portion of the sample and the second portion of the sample based on a relationship between the hemoglobin concentration and the mean corpuscular hemoglobin.

[0023] In some applications, determining a parameter of the sample comprises determining a count of a component of the blood, wherein said component is selected from the group consisting of: red blood cells, red blood cells of a given type, white blood cells, white blood cells of a given type, circulating tumor cells, platelets, platelets of a given type, bacteria, pathogens, pathogens of a given type, reticulocytes, and Howell-Jolly bodies.

[0024] In some applications, determining a parameter of the sample comprises determining a concentration of a component of the blood, wherein said component is selected from the group consisting of: red blood cells, red blood cells of a given type, white blood cells, white blood cells of a given type, circulating tumor cells, platelets, platelets of a given type, bacteria, pathogens, pathogens of a given type, reticulocytes, and Howell-Jolly bodies.

[0025] In some applications, determining a parameter of the sample comprises determining the hematocrit of the sample.

[0026] In some applications, measuring the hemoglobin concentration comprises measuring the hemoglobin concentration in a first portion of the blood sample, and measuring the mean corpuscular hemoglobin in the blood sample comprises measuring the mean corpuscular hemoglobin in a second portion of the blood sample, wherein said second portion of the blood sample is diluted relative to the first portion of the blood sample.

[0027] In some applications, determining a parameter of the blood sample comprises determining a normalization factor by determining a property of a first portion of the sample portion, as a reference that can be used to correct measurements in a second portion.

[0028] In some applications, determining a parameter of the blood sample comprises determining the red blood cell count in the sample by dividing the concentration of hemoglobin by the mean corpuscular hemoglobin.

[0029] In some applications, determining a parameter of the blood sample further comprises determining a count of one or more components in the sample based on the red blood cell count in the sample.

[0030] In some applications, determining a count of one or more components in the sample comprises:

[0031] - determining a ratio of the red blood cell count to the count of one or more components in the sample portion by analyzing the microscopic image of the sample portion; and

[0032] - determining the count of one or more components based on the red blood cell count in the sample, and the ratio of the red blood cell count to the count of one or more components in the sample portion.

[0033] According to some applications of the application, there is further provided an apparatus for a blood sample, the apparatus comprising:

[0034] - at least one computer processor configured to:

[0035] • measure a hemoglobin concentration in at least a portion of the blood sample by performing a first measurement of the blood sample;

[0036] • measure a mean corpuscular hemoglobin in the blood sample by performing a second measurement of the blood sample; and

[0037] • determine a parameter of the blood sample based on a relationship between the hemoglobin concentration and the mean corpuscular hemoglobin.

[0038] According to some applications of the application, there is further provided a computer software product for a blood sample, the computer software product comprising a non-transitory computer readable medium having stored thereon program instructions that, when read by a computer, cause the computer to perform the steps of:

[0039] - measuring a hemoglobin concentration in at least a portion of the blood sample by performing a first measurement of the blood sample;

[0040] - measuring a mean corpuscular hemoglobin in the blood sample by performing a second measurement of the blood sample; and

[0041] - determining a parameter of the blood sample based on a relationship between the hemoglobin concentration and the mean corpuscular hemoglobin.

[0042] According to some applications of the application, there is further provided a method for a blood sample, the method comprising:

[0043] - measuring a hematocrit of the blood sample by performing a first measurement of the blood sample;

[0044] - measuring a mean corpuscular volume of the blood sample by performing a second measurement of the blood sample; and

[0045] - determining a parameter of the blood sample based on a relationship between the hematocrit and the mean corpuscular volume.

[0046] In some applications, determining the parameter of the blood sample comprises normalizing the first and second measurements relative to each other based on the relationship between the hematocrit and the mean corpuscular volume.

[0047] In some applications, performing the first measurement of the blood sample comprises performing a measurement of the blood sample selected from the group consisting of: an ultrasonic measurement and an impedance measurement.

[0048] In some applications, performing the first measurement of the blood sample comprises centrifuging the blood sample.

[0049] In some applications, performing the second measurement comprises performing the second measurement by analyzing a microscopic image of the portion of the blood sample.

[0050] In some applications, measuring the hematocrit comprises measuring the hematocrit of a first portion of the blood sample, measuring the mean corpuscular volume of the blood sample comprises measuring the mean corpuscular volume of a second portion of the blood sample, and determining the parameter of the sample comprises determining a relationship between the first portion of the sample and the second portion of the sample based on a relationship between the hematocrit and the mean corpuscular volume.

[0051] In some applications, determining the parameter of the sample comprises determining a count of a blood component selected from the group consisting of red blood cells, red blood cells of a given type, white blood cells, white blood cells of a given type, circulating tumor cells, platelets, platelets of a given type, bacteria, pathogens, pathogens of a given type, reticulocytes, and Howell-Jolly bodies.

[0052] In some applications, determining the parameter of the sample comprises determining a concentration of a blood component selected from the group consisting of red blood cells, red blood cells of a given type, white blood cells, white blood cells of a given type, circulating tumor cells, platelets, platelets of a given type, bacteria, pathogens, pathogens of a given type, reticulocytes, and Howell-Jolly bodies.

[0053] In some applications, measuring the hematocrit comprises measuring the hematocrit of a first portion of the blood sample, and measuring the mean corpuscular volume of the blood sample comprises measuring the mean corpuscular volume of a second portion of the blood sample, wherein the second portion of the blood sample is diluted relative to the first portion of the blood sample.

[0054] In some applications, determining the parameter of the blood sample comprises determining a normalization factor by determining a property of a first portion of the same portion, which is used as a reference that can correct measurements of a second portion.

[0055] In some applications, determining the parameter of the blood sample comprises determining a count of red blood cells in the sample by dividing the hematocrit by the mean corpuscular volume.

[0056] In some applications, determining the parameter of the blood sample further comprises determining a count of one or more components in the sample based on the count of red blood cells in the sample.

[0057] In some applications, determining the count of one or more components in the sample comprises:

[0058] - determining a ratio of the count of red blood cells to the count of one or more components in the sample portion by analyzing the microscopic image of the sample portion; and

[0059] - determining the count of one or more components based on the count of red blood cells in the sample and the ratio of the count of red blood cells to the count of one or more components in the sample portion.

[0060] According to some applications of the application, there is further provided an apparatus for a blood sample, the apparatus comprising:

[0061] - at least one computer processor configured for:

[0062] • measuring the hematocrit of the blood sample by performing a first measurement on the blood sample;

[0063] • measuring the mean corpuscular volume in the blood sample by performing a second measurement on the blood sample; and

[0064] • determining a parameter of the blood sample based on a relationship between the hematocrit and the mean corpuscular volume.

[0065] According to some applications of the application, there is further provided a computer software product for a blood sample, the computer software product comprising a non-transitory computer readable medium having stored thereon program instructions that, when executed by a computer, cause the computer to perform the steps of:

[0066] - measuring the hematocrit of the blood sample by performing a first measurement on the blood sample;

[0067] - measuring the mean corpuscular volume in the blood sample by performing a second measurement on the blood sample; and - determining a parameter of the blood sample based on a relationship between the hematocrit and the mean corpuscular volume.

[0068] According to some applications of the application, there is further provided a method for a first portion of a blood sample and a second portion of the blood sample, wherein the second portion of the blood sample is diluted relative to the first portion of the blood sample, the method comprising:

[0069] - measuring the relative amounts of the first and second components in the first portion of the blood sample;

[0070] - measuring a measured variable of the second portion of the blood sample; and

[0071] - determining a parameter of the blood sample based on a relationship between the relative amounts of the first and second components in the first portion of the blood sample and the measured variable of the second portion of the blood sample.

[0072] In some applications, measuring the relative amounts of the first and second components in the first portion of the blood sample comprises analyzing a microscopic image of the first portion of the blood sample.

[0073] In some applications, measuring the relative amounts of the first and second components in the first portion of the blood sample comprises measuring the relative amounts of at least two components in the first portion of the blood sample, said two components being selected from the group consisting of: all leukocyte types, neutrophils, eosinophils, basophils, lymphocytes, monocytes, and leukocyte precursors.

[0074] In some applications, measuring the relative amounts of the first and second components in the first portion of the blood sample comprises measuring the relative amounts of at least two components in the first portion of the blood sample, said two components being selected from the group consisting of: red blood cells, reticulocytes, inclusions, red blood cells with a given morphology, and Howell-Jolly bodies.

[0075] In some applications, measuring the relative amounts of the first and second components in the first portion of the blood sample comprises measuring the relative amounts of a given type of platelets in the first portion of the blood sample.

[0076] In some applications, measuring the measured variable in the second portion of the sample comprises measuring the absolute amount of a given type of cells in the second portion of the blood sample.

[0077] In some applications, measuring the measured variable in the second portion of the sample comprises measuring the concentration of a given component in the second portion of the blood sample.

[0078] In some applications, measuring the measured variable in the second portion of the sample comprises performing a batch level measurement on the second portion of the blood sample.

[0079] According to some applications of the application, there is further provided an apparatus for a blood sample, the apparatus comprising:

[0080] - at least one computer processor configured for:

[0081] • measuring the relative amounts of the first and second components in the first portion of the blood sample;

[0082] • measuring the measured variable in the second portion of the blood sample; and

[0083] • determining a parameter of the blood sample based on a relationship between the relative amounts of the first and second components in the first portion of the blood sample and the measured variable in the second portion of the blood sample.

[0084] According to some applications of the application, there is further provided a computer software product for a blood sample, the computer software product comprising a non-transitory computer readable medium having stored thereon program instructions that, when executed by a computer, cause the computer to perform the steps of:

[0085] • measuring the relative amounts of the first and second components in the first portion of the blood sample;

[0086] - measuring the measured variable in the second portion of the blood sample; and

[0087] - determining a parameter of the blood sample based on a relationship between the relative amounts of the first and second components in the first portion of the blood sample and the measured variable in the second portion of the blood sample.

[0088] According to some applications of the application, there is further provided a method for a biological sample, the method comprising:

[0089] - measuring a measured variable at a bulk level of the sample by performing a first measurement on the sample;

[0090] - measuring a measured variable at a cellular level of the sample by performing a second measurement on the sample; and - determining a parameter of the sample based on a relationship between the measured variable at the bulk level and the measured variable at the cellular level.

[0091] In some applications, determining a parameter of the blood sample comprises normalizing the first and second measurements with respect to the batch based on a relationship between the measured variable at the bulk level and the measured variable at the cellular level.

[0092] In some applications, measuring the measured variable at the bulk level comprises determining an optical density of a given component in the sample.

[0093] In some applications, measuring the measured variable at the cellular level comprises analyzing a microscopic image of the sample.

[0094] In some applications, performing the first measurement on the sample comprises performing the first measurement on the sample using a first set of measurement conditions, performing the second measurement on the sample comprises performing the second measurement on the sample using a second set of measurement conditions, and determining a parameter of the sample comprises determining a relationship between the measurement conditions used to perform the first and second measurements based on a relationship between the measured variable at the bulk level and the measured variable at the cellular level.

[0095] In some applications, performing the first measurement comprises performing the first measurement on a first portion of the sample, and performing the second measurement comprises performing the second measurement on the first portion of the sample.

[0096] In some applications, performing the first measurement comprises performing a first measurement on a first portion of the sample, and performing the second measurement comprises performing a second measurement on a second portion of the sample different from the first portion of the sample. In some applications, determining the parameter of the sample comprises determining a relationship between the first portion of the sample and the second portion of the sample based on a relationship between the measured variable at the bulk level and the measured variable at the cellular level. In some applications, performing the second measurement on the second portion of the sample comprises performing a second measurement on the second portion of the sample, wherein the second portion of the sample is diluted relative to the first portion of the sample. In some applications, determining the parameter of the sample comprises determining a property of the first portion of the sample portion, determining a normalization factor as a reference that can correct the measurement in the second portion. In some applications, determining the parameter of the sample comprises determining a dilution ratio by which the second portion of the sample is diluted relative to the first portion of the sample.

[0097] In some applications, the biological sample comprises a blood sample, and determining the parameter of the sample comprises determining a parameter of the blood sample.

[0098] In some applications:

[0099] - measuring the measured variable at the bulk level of the sample comprises measuring the hematocrit of the blood sample;

[0100] - measuring the measured variable at the cellular level of the sample comprises measuring the mean corpuscular volume of the blood sample;

[0101] and

[0102] - determining the parameter of the sample comprises determining the parameter of the sample based on a relationship between the hematocrit and the mean corpuscular volume.

[0103] In some applications:

[0104] - measuring the measured variable at the bulk level of the sample comprises measuring the hemoglobin concentration in at least a portion of the blood sample;

[0105] - measuring the measured variable at the cellular level of the sample comprises measuring the mean corpuscular hemoglobin of the blood sample; and

[0106] - determining the parameter of the sample comprises determining the parameter of the sample based on a relationship between the hemoglobin concentration and the mean corpuscular hemoglobin.

[0107] According to some applications of the application, there is further provided an apparatus for a biological sample, the apparatus comprising:

[0108] - at least one computer processor configured for:

[0109] • measuring a measured variable at a bulk level of the sample by performing a first measurement on the sample;

[0110] • measuring the measured variable at the cell level of the sample by performing a second measurement on the sample; and

[0111] • determining the parameter of the sample based on the relationship between the measured variable at the bulk level and the measured variable at the cell level.

[0112] According to some applications of the application, there is further provided a computer software product for a biological sample, the computer software product comprising a non-transitory computer readable medium having stored therein program instructions which, when read by a computer, cause the computer to perform the steps of:

[0113] - measuring the measured variable at the bulk level of the sample by performing a first measurement on the sample;

[0114] - measuring the measured variable at the cell level of the sample by performing a second measurement on the sample; and - determining the parameter of the sample based on the relationship between the measured variable at the bulk level and the measured variable at the cell level.

[0115] According to some applications of the application, there is further provided a method for a biological sample, the method comprising:

[0116] - performing first and second optical measurements on the sample using one or more optical measurement devices under a plurality of sets of measurement conditions which are different from each other;

[0117] - measuring the measured variable of the sample based on the first optical measurement;

[0118] - measuring the measured variable of the sample based on the second optical measurement; and

[0119] - determining the relationship between the measurement conditions of the one or more optical measurement devices used to perform the first and second optical measurements based on the relationship between the measured variable measured based on the first optical measurement and the measured variable measured based on the second optical measurement.

[0120] In some applications, the biological sample comprises a blood sample, and performing the first and second optical measurements on the sample comprises performing the first and second optical measurements on the blood sample.

[0121] In some applications:

[0122] - performing the first and second optical measurements on the sample comprises performing the first and second optical measurements on a plurality of portions of the sample, wherein the plurality of portions of the sample are disposed in a plurality of portions of one or more sample chambers having a plurality of dimensions; and

[0123] - determining the relationship between the measurement conditions of the one or more optical measurement devices includes determining a relationship between the dimensions of a plurality of portions of the one or more sample chambers, wherein the optical measurement devices are used to perform the first and second optical measurements.

[0124] In some applications, performing the first and second optical measurements on the sample includes performing at least one of the first and second optical measurements by acquiring an image of at least a portion of the sample, and determining the relationship between the measurement conditions of the one or more optical measurement devices includes determining a field of view of the image, wherein the optical measurement devices are used to perform the first and second optical measurements.

[0125] In some applications, performing the first and second optical measurements on the sample includes performing at least one of the first and second optical measurements by acquiring an image of at least a portion of the sample, and determining the relationship between the measurement conditions of the one or more optical measurement devices includes determining a magnification level of the image, wherein the optical measurement devices are used to perform the first and second optical measurements.

[0126] In some applications:

[0127] - measuring the measured variable of the sample based on the first optical measurement includes measuring a given measured variable of the sample based on the first optical measurement;

[0128] - measuring the measured variable of the sample based on the second optical measurement includes measuring the same given measured variable of the sample based on the second optical measurement; and

[0129] - determining the relationship between the measurement conditions of the one or more optical measurement devices (used to perform the first and second optical measurements) includes determining the relationship between the measurement conditions of the one or more optical measurement devices (used to perform the first and second optical measurements) based on a relationship between the given measured variable measured based on the first optical measurement and the given measured variable measured based on the second optical measurement.

[0130] In some applications, performing the first optical measurement includes performing the first optical measurement using a given optical measurement device, and performing the second optical measurement includes performing the second optical measurement using the same given optical measurement device.

[0131] In some applications, performing the first optical measurement includes performing the first optical measurement using a first optical measurement device, and performing the second optical measurement includes performing the second optical measurement using a second optical measurement device that is different from the first optical measurement device.

[0132] In some applications:

[0133] - performing the first optical measurement comprises performing the first optical measurement using a first optical measurement device configured for measuring a parameter of one or more components in the sample, the parameter being selected from the group consisting of: optical absorption, transmission, fluorescence, and luminescence; and

[0134] - performing the second optical measurement comprises performing the second optical measurement using a microscope configured for acquiring a microscopic image of the sample.

[0135] In some applications:

[0136] - measuring the measured variable of the sample based on the first optical measurement comprises measuring a first measured variable of the sample based on the first optical measurement; and

[0137] - measuring the measured variable of the sample based on the second optical measurement comprises measuring a second measured variable of the sample different from the first measured variable based on the second optical measurement; and

[0138] - determining the relationship between the measurement conditions of the one or more optical measurement devices used for performing the first and second optical measurements comprises determining the relationship between the measurement conditions of the one or more optical measurement devices used for performing the first and second optical measurements based on the relationship between the first and second measured variables.

[0139] In some applications, measuring the first measured variable comprises measuring the measured variable at a bulk level of the sample, and measuring the second measured variable comprises measuring the measured variable at a cellular level of the sample.

[0140] According to some applications of the present application, there is further provided an apparatus for biological samples, the apparatus comprising:

[0141] - at least one computer processor configured for:

[0142] • performing first and second optical measurements of the sample using one or more optical measurement devices under a plurality of sets of measurement conditions different from each other;

[0143] • measuring a measured variable of the sample based on the first optical measurement;

[0144] • measuring the measured variable of the sample based on the second optical measurement; and

[0145] • determining a relationship between the measurement conditions of the one or more optical measurement devices used for performing the first and second optical measurements based on a relationship between the measured variable measured based on the first optical measurement and the measured variable measured based on the second optical measurement.

[0146] According to some applications of the application, there is further provided a computer software product for a biological sample, the computer software product comprising a non-transitory computer readable medium having stored thereon program instructions that, when executed by a computer, cause the computer to perform the steps of:

[0147] - performing first and second optical measurements of the sample using one or more optical measurement devices under a plurality of sets of measurement conditions that are different from each other;

[0148] - measuring a measured variable of the sample based on the first optical measurement;

[0149] - measuring the measured variable of the sample based on the second optical measurement; and

[0150] - determining a relationship between the measurement conditions of the one or more optical measurement devices used to perform the first and second optical measurements based on a relationship between the measured variable measured based on the first optical measurement and the measured variable measured based on the second optical measurement.

[0151] According to some applications of the application, there is further provided a method for a biological sample, the method comprising:

[0152] - performing first and second optical measurements of the sample using one or more optical measurement devices under a plurality of sets of measurement conditions that are different from each other;

[0153] - measuring a measured variable of the sample based on the first optical measurement;

[0154] - measuring the measured variable of the sample based on the second optical measurement;

[0155] - normalizing the measured variable measured based on the first optical measurement and the measured variable measured based on the second optical measurement with respect to each other; and

[0156] - determining a parameter of the sample based on at least one of the normalized measured variable measured based on the first optical measurement and the normalized measured variable measured based on the second optical measurement.

[0157] According to some applications of the application, there is further provided an apparatus for a biological sample, the apparatus comprising:

[0158] - at least one computer processor configured to:

[0159] • perform first and second optical measurements of the sample using one or more optical measurement devices under a plurality of sets of measurement conditions that are different from each other;

[0160] • measure a measured variable of the sample based on the first optical measurement;

[0161] • measuring a measured variable of the sample based on the second optical measurement;

[0162] • normalizing the measured variable measured on the basis of the first optical measurement and the measured variable measured on the basis of the second optical measurement with respect to each other; and

[0163] • determining a parameter of the sample based on at least one of the normalized measured variable measured on the basis of the first optical measurement and the normalized measured variable measured on the basis of the second optical measurement.

[0164] According to some applications of the application, there is further provided a computer software product for a biological sample, the computer software product comprising a non-transitory computer readable medium having stored therein program instructions which, when read by a computer, cause the computer to perform the steps of:

[0165] - performing first and second optical measurements of the sample using one or more optical measurement devices under a plurality of sets of measurement conditions different from each other;

[0166] - measuring a measured variable of the sample based on the first optical measurement;

[0167] - measuring a measured variable of the sample based on the second optical measurement;

[0168] - normalizing the measured variable measured on the basis of the first optical measurement and the measured variable measured on the basis of the second optical measurement with respect to each other; and

[0169] - determining a parameter of the sample based on at least one of the normalized measured variable measured on the basis of the first optical measurement and the normalized measured variable measured on the basis of the second optical measurement.

[0170] The application will be more fully understood from the following detailed description of embodiments thereof, taken together with the drawings in which: BRIEF DESCRIPTION OF DRAWINGS

[0171] Figure 1 a block diagram of components of a biological sample analysis system according to some applications of the application;

[0172] Figure 2 a schematic illustration of a sample carrier according to some applications of the application;

[0173] Figure 3 a flowchart of steps of performing calculations according to some applications of the application;

[0174] Figure 4 a flowchart of steps of performing calculations according to some applications of the application;

[0175] Figure 5A flowchart of the steps performed in carrying out the operations for some applications in accordance with the present application;

[0176] Figure 6 A flowchart of the steps performed in carrying out the operations for some applications in accordance with the present application;

[0177] Figure 7 A flowchart of the steps performed in carrying out the operations for some applications in accordance with the present application; and

[0178] Figure 8 A schematic cross-sectional view of a sample carrier defining stepped height variations for some applications in accordance with the present application. DETAILED DESCRIPTION

[0179] Reference is now made to Figure 1 which is a block diagram showing components of a biological sample analysis system 20 for some applications in accordance with the present application. Typically, a biological sample (e.g., a blood sample) is placed on a sample carrier 22. When the sample is disposed on the sample carrier, one or more optical measurement devices 24 are used to make optical measurements of the sample. For example, the optical measurement devices can include microscopes (e.g., digital microscopes), spectrometers, photometers, spectrophotometers, cameras, spectral cameras, hyperspectral cameras, fluorometers, spectrofluorometers, and / or photodetectors (e.g., photodiodes, photoresistors, and / or phototransistors). For some applications, the optical measurement devices include dedicated light sources (e.g., light emitting diodes, incandescent light sources, etc.) and / or optical elements (e.g., lenses, diffusers, filters, etc.) for manipulating light collection and / or light emission. For some applications, a microscope system is used that is generally similar to the microscope system described by Greenfield in US 2014 / 0347459, which is incorporated herein by reference.

[0180] The computer processor 28 typically receives and processes optical measurements performed by the optical measurement device. More typically, the computer processor controls acquisition of optical measurements performed by one or more optical measurement devices. The computer processor is in communication with the memory 30. A user (e.g., a laboratory technician) sends instructions to the computer processor through the user interface 32. For some applications, the user interface includes a keyboard, a mouse, a joystick, a touch screen device (e.g., a smart phone or tablet), a touch pad, a trackball, a voice command interface, and / or other types of user interfaces known in the art. Typically, the computer processor generates output through the output device 34. More typically, the output device includes a display, e.g., a detector meter, and the output includes output displayed on the display. For some applications, the processor generates output on different types of visual, textual, graphical, tactile, audio, and / or video output devices (e.g., a speaker, headphones, a smart phone or tablet). For some applications, the user interface 32 serves as an input interface and an output interface, e.g., it serves as an input / output interface. For some applications, the processor generates output on a computer readable medium (e.g., a non-transitory computer readable medium), e.g., a disk or a portable USB drive, or on a printer.

[0181] Reference is now made to Figure 2 which is a schematic illustration of a sample carrier 22, in accordance with some applications of the present application. For some applications, the sample carrier includes a source sample portion chamber 40, and a dilution sample portion chamber 42. Typically, the chambers 40 and 42 are filled through respective access ports 44 and 46.

[0182] For some applications, the dilution sample portion chamber 42 is filled with a second portion 50 of the biological sample (e.g., a portion of the blood sample), which is diluted relative to the first portion 48 of the sample placed in the source sample portion chamber 40. For example, a portion of the sample can be diluted in order to identify and / or count components of the sample that can not be readily identifiable and / or countable in an undiluted portion of the sample. For some applications, the dilution portion comprises a staining substance. For example, the dilution portion can be prepared using the techniques described by Pollak in US 2015 / 0316477, which is incorporated by reference herein, and which describes methods of preparation of a blood sample for analysis involving a dilution step that facilitates identification and / or counting of components in a microscopic image of the sample. Typically, in such applications, although the degree of dilution is typically set as part of the protocol, small variations in the degree of dilution can lead to corresponding errors in the determination of the quantity of different components and / or analytes in the sample. According to some applications of the present application, two different measurements are performed on the first portion 48 of the sample placed in chamber 40 (i.e., the source sample portion) and on the second portion 50 of the sample placed in chamber 42 (i.e., the dilution sample portion), which is diluted relative to the first portion (i.e., the dilution sample portion). For some applications, based on the measurements, the dilution factor (i.e., the dilution ratio, and / or the degree of dilution of the second portion relative to the first portion) is determined. Typically, a normalization factor is determined, which is a property of the source sample portion (e.g., the number of red blood cells per unit area or the number of red blood cells per unit volume in the source sample portion) that is related to the other measurements. More typically, based on the normalization factor, the measured variable in the sample (e.g., in the source sample portion) is determined, as described in further detail below.

[0183] For some applications, the methods described herein are performed on a source sample portion and a dilution sample portion, which are not portions of a sample placed in separate chambers of a single sample carrier, as shown in Figure 2 For some applications, the methods described herein are performed on a first and a second portion of a sample, which are not diluted relative to each other (modified as appropriate). For some such applications, each measurement (e.g., each optical measurement) performed on the first and second portions using the techniques described herein is normalized relative to the other.

[0184] For some applications, the source sample portion 48 placed in the source sample portion chamber is a native, undiluted biological fluid (e.g., a blood sample or a urine sample), or is a sample that has been altered in some way, e.g., including one or more of dilution (e.g., dilution in a controlled manner), addition of an ingredient or reagent, or fractionation. The diluted sample portion 50 placed in the diluted sample portion chamber is typically diluted relative to the sample portion placed in the source sample portion chamber. For example, the diluent can include a pH buffer, a stain, a fluorescent stain, an antibody, a globin, a lysing agent, etc.

[0185] In general, the diluted sample portion 50 is tested to provide a plurality of measurements, with the assumption that the measurements are in good relative agreement with one another. More generally, the source sample portion 48 is tested, where the source sample portion produces a measurement that is comparable to at least one measurement performed on the diluted sample portion. For some applications, at least one measurement performed on the diluted sample portion 50 is normalized by using a measurement measured by and / or derived from the portions 48 and 50.

[0186] For example, a blood sample can be diluted using the dilution techniques described by Pollak in US 2015 / 0316477, which is incorporated by reference herein, and the smear can be appropriately stained and imaged using a microscope system (which can be manual or automated). For some applications, the microscope system is a type of optical measurement device 24, as described above with reference to Figure 1

[0187] For some applications, the microscopic images are analyzed (e.g., manually or using a computer processor running appropriate computer software) to identify different blood cells. However, an error of 10% in the dilution factor can directly correspond to a 10% error in the count of red blood cells per unit volume (e.g., per microliter) of, for example, blood. Such an error in dilution can arise from a variety of sources. Illustrative examples of sources of such errors (which are not intended to limit the scope of the application) include inaccuracy or error in pipetting, inaccuracy or error in calibration, inaccuracy or error in mixing, etc.

[0188] ​For some applications, the methods described herein are used to account for at least some variation in dilution by measuring a source sample portion (e.g., an undiluted blood sample portion) that is used to extract a diluted sample portion. Such a measurement is typically equivalent to at least one measurement taken on the diluted sample portion. For example, a measurement taken on the source sample portion can include a measurement of hemoglobin content, white blood cell content, red blood cell content, hematocrit, content of specific white blood cell types, platelet content, and / or any measured variable measured on or inferable from the diluted sample portion. For some applications, a normalization factor is determined that is a property of the source sample portion related to other measurements (e.g., number of red blood cells per unit area or number of red blood cells per unit volume in the source sample portion). Typically, the measured variable in the sample (e.g., the source sample portion) is measured based on the normalization factor, as described in more detail below. For some applications, multiple measurements are taken on the source sample portion 48 (e.g., two or more of the measurements described above), and a normalization factor (e.g., dilution factor) is determined based on the multiple measurements. Typically, in such cases, the normalization factor is determined based on different portions of the data using statistical methods (e.g., averaging, regression, curve fitting, or other techniques known in the art). For some applications, the use of a large number of analytical measurements increases the accuracy of the normalization relative to the use of only a single measurement, as described above.

[0189] Typically, the methods described above are performed on two or more portions of a sample at different dilution levels (i.e., a source sample portion and a diluted sample portion), as described above, whereby the amount or concentration of a different component in one sample portion is determined based on the dilution factor between the two sample portions. For example, the methods described can be used to determine the amount or concentration of different blood components in a whole blood count test performed on a diluted blood sample portion.

[0190] For some applications, the dilution factor (i.e., the dilution ratio (e.g., 1 : 100) and / or the degree of dilution of the second portion relative to the first portion) is determined. For some applications, the same measured variable is measured in both the diluted and source sample portions. For example, the count per unit volume of a component, the concentration of a component, and / or the optical density of a component can be measured. The dilution factor of the diluted sample portion relative to the source sample portion is derived from the ratio of the measured variable measured in both sample portions (e.g., the ratio of the count per unit volume of a component, the concentration of a component, and / or the optical density of a component measured in both sample portions). The dilution factor is typically used to determine a parameter related to one or more other components (e.g., count per unit volume, concentration, and / or optical density).

[0191] As noted above, generally, a normalization factor is determined, which is a property of the source sample portion (e.g., red blood cell number / unit area or red blood cell number / unit volume in the source sample portion) that is related to other measurements. For some applications, one measured variable is measured in the diluted sample portion, and a different measured variable is measured in the diluted sample portion. For example, hemoglobin concentration (Hb) can be measured in the source blood sample portion, and mean corpuscular hemoglobin (MCH) can be measured in the diluted blood sample portion. Alternatively, hematocrit can be measured in the source blood sample portion, and mean corpuscular volume (MCV) can be measured in the diluted blood sample portion. Generally, a relationship between the two measurements is determined, and based on the relationship, the concentration, or number / unit volume, of the reference component in the source sample portion is inferred. Thereafter, parameters related to one or more other components (e.g., number / unit volume, concentration, and / or optical density) are determined in association with the ratio of the other components to the reference component. The techniques described above are more easily understood by the following examples.

[0192] For some applications (e.g., with respect to a complete blood count), total hemoglobin concentration ("Hb") is determined by performing a measurement on the undiluted sample, e.g., using an optical density measurement performed on undiluted blood. Generally, the measurement is performed using an optical measurement device 24, such as a spectrometer, a spectrophotometer, a camera, a spectral camera, a hyperspectral camera. Figure 1 Mean corpuscular hemoglobin (MCH) is determined using the diluted blood sample portion. The optical density measurement can be performed at the cellular level (i.e., with respect to individual cells), for example. A microscope can be used as the optical measurement device 24, for example. Figure 1Cells can be imaged using bright-field imaging at violet or green wavelengths. For some applications, the red blood cell count per unit volume ("RBC") in the source sample fraction is inferred by dividing the hemoglobin concentration by the mean erythrocyte hemoglobin (since RBC = Hb / MCH). For some applications, the counts of other components in the source sample fraction are determined based on the RBC count per unit volume. For example, using a microscopic image of a diluted sample fraction, the ratio of the counts of other blood components (e.g., given types of red blood cells, white blood cells, circulating tumor cells, platelets, bacteria, pathogens, reticulocytes, and Howell-Jolly bodies) to the red blood cell count can be determined. Alternatively or additionally, the ratio of the counts of other blood components to the red blood cell count can be determined using an undiluted sample fraction, wherein the microscopic image forms a monolayer with a sufficiently low cell density for identifying a single component within the monolayer (e.g., by placing the fraction in a sample chamber at a relatively low height). By multiplying this ratio by the red blood cell count, the absolute counts of other components in the source sample portion are determined. For example, once based on (WBC / RBC)... 稀释 To determine the ratio of white blood cells to red blood cells in a diluted sample, the WBC count is used. 计数 =(WBC / RBC) 稀释 ×RBC=(WBC / RBC) 稀释 ×Hb / MCH, calculates the white blood cell count per unit volume ("WBC count"). 计数 For some applications, the hematocrit in a sample is determined based on the measured red blood cell count. For other applications, the mean corpuscular volume (MCV) can be measured relative to a diluted portion of the sample, and the hematocrit can be determined by multiplying the MCV by the red blood cell count.

[0193] For some applications (e.g., in the case of whole blood counting), such as the microhematocrit method (in which a certain amount of blood is centrifuged) or using ultrasound and / or impedance measurement, the hematocrit ("HCT") is determined on an undiluted sample. Using a diluted portion of the blood sample, the mean erythrocyte volume ("MCV") is determined. For example, a microscope can be used as an optical measuring device 24 to image the red blood cells (…). Figure 1The image can be used to derive the mean erythrocyte volume (RBC). For some applications, the red blood cell count per unit volume ("RBC") in the source sample portion is inferred by dividing the hematocrit by the mean erythrocyte volume (since RBC = HCT / MCV). For some applications, the counts of other components in the source sample portion are determined based on the RBC per unit volume. For example, using a microscopic image of a diluted sample portion, the ratio of the counts of other blood components (e.g., given types of red blood cells, white blood cells, circulating tumor cells, platelets, bacteria, pathogens, reticulocytes, and Howell-Jolly bodies) to the red blood cell count can be determined. Alternatively or additionally, an undiluted sample portion can be used to determine the ratio of the counts of other blood components to the red blood cell count, wherein the microscopic image forms a monolayer with a sufficiently low cell density for identifying a single component within the monolayer (e.g., by placing the portion in a sample chamber at a relatively low height). The absolute count of other components in the source sample portion is determined by multiplying this ratio by the red blood cell count. For example, once according to WBC / RBC) diluted To determine the ratio of white blood cells to red blood cells in a diluted sample, the WBC count is used. eount =(WBC / RBC) dilutsd ×RBC=(WBC / RBC) diluted ×HCT / MCV, calculate the white blood cell count per unit volume (WBC) in the source sample. count For some applications, the concentration of hemoglobin in a sample is determined based on the measured red blood cell count. For example, mean corpuscular hemoglobin (MCH) can be measured relative to a diluted portion of the sample, and the concentration of hemoglobin can be determined by multiplying MCH by the red blood cell count.

[0194] For some applications (e.g., with respect to a complete blood count), the total white blood cell count per unit volume in the source sample portion is determined. The total white blood cell count per unit volume can be determined, for example, by (a) lysing the red blood cells in the sample (so that the red blood cells do not scatter light); (b) imaging the sample using a DNA-specific stain (e.g., methylene blue) that has a high absorption wavelength at which the absorbance of hemoglobin from the red blood cells is low; and (c) measuring the absorbance at these wavelengths. For some applications, the counts of other components in the source sample portion are determined based on the white blood cell count per unit volume in the source sample portion. For example, using microscopic images of a diluted sample portion, the ratio of the counts of other blood components (e.g., red blood cells, red blood cells of a given type, white blood cells of a given type, circulating tumor cells, platelets, platelets of a given type, bacteria, pathogens, pathogens of a given type, reticulocytes, and Howell-Jolly bodies) to the basophil count can be determined. By multiplying this ratio by the white blood cell count, the absolute counts of the other components in the source sample portion are determined.

[0195] For some applications, the even source sample portion 48 is not a native biological sample, but is itself diluted. For some such applications, the counts and / or concentrations of components in a native sample from which the source sample portion was generated are derived. For example, a native blood sample can be diluted in a controlled and precise manner to generate the source sample portion, and the dilution factor of the dilution step is known exactly. The source sample portion is then used to generate a diluted sample portion 50 as described above, and the counts per unit volume and / or concentrations of some blood components in the source sample portion are derived as described above. Based on the counts per unit volume and / or concentrations of the blood components in the source sample portion, the counts per unit volume and / or concentrations of these components in the native sample are derived.

[0196] For some applications, a native sample is diluted to generate a source sample portion 48, which is further diluted to generate a diluted sample portion 50. Based on the parameters determined for each of the source sample portion and the diluted sample portion, the parameters of the native sample are extrapolated without directly estimating the dilution factor. For example, the ratio of white blood cells to red blood cells can be determined using microscopic images of the diluted sample portion, while the ratio of basophils to white blood cells can be determined for the source sample portion, as described above. In addition, the red blood cell count per unit volume of the native sample can be determined. Thus, the basophil count per unit volume of the native sample can be determined using the red blood cell count per unit volume of the native sample, in combination with the ratios.

[0197] Referring again to Figure 2For some applications, the techniques described herein are implemented using a carrier 22 having at least two chambers for each patient (or source), as described above. Typically, the source sample portion chamber 40 is configured for testing a small volume, e.g., 1 to 30 microliters, of blood, so that, for example, more blood need not be drawn. More typically, the source sample portion chamber and the diluted sample portion chamber are in close proximity to each other, e.g., as shown in Figure 2 FIG. 2, by being disposed on a single sample carrier. For some applications, the source and diluted sample chambers being in close proximity to each other advantageously reduces the risk of source sample and diluted sample mix-up.

[0198] As described above, for some applications, the methods described herein are implemented for both the source sample portion and the diluted sample portion without placing the portions in separate chambers of a single sample carrier, as shown in Figure 2 FIG. 2. For some applications, the methods described herein are implemented for first and second portions of a sample without diluting the portions relative to each other (modified as appropriate). For such applications, the various measurements (e.g., optical measurements) implemented on the first and second portions using the techniques described herein are normalized relative to each other.

[0199] Although some of the examples described above are described with reference to certain measurements implemented on source and diluted sample portions of a blood sample, the scope of the present application generally includes combinations of measurements (e.g., optical measurements) implemented on a sample (and / or portions thereof) from which parameters of the sample are derived, as described with reference to the flowchart shown in Figures 3-7 FIG. 3.

[0200] Reference is now made to Figure 3which is a flow chart showing the steps of performing the operations according to some applications of the present application. According to some applications of the present application, two or more measurements are performed on a biological sample, which are typically optical measurements. Typically, the biological sample is a blood sample. For some applications, in a first step 60, a bulk level of a measured variable of the sample is measured by performing a first measurement on the sample. More typically, in a second step 62, a cell level of a measured variable of the sample is measured by performing a second measurement on the sample. For the purposes of the present application, the term "cell level of a measured variable" is understood to refer to a measured variable related to one or more parameters of individual cells or other non-dissolved components in the sample, such as mean corpuscular volume, mean corpuscular hemoglobin, mean platelet volume and / or mean platelet red cell distribution width, etc. Measuring a cell level of a measured variable typically involves a first step of identifying individual cells or other non-dissolved components in the sample (such as identifying the components in a microscopic image), and a second step of identifying parameters of such individual identified components. For some applications, the cell level of a measured variable is measured by analyzing one or more microscopic images of the sample. For the purposes of the present application, the term "bulk level of a measured variable" is understood to refer to a measured variable related to a parameter of the sample as a whole, and which does not require two steps of identifying individual cells or other non-dissolved components in the sample and identifying parameters of such individual identified components. For example, such measured variables can include optical density of a given component (which is measured by performing a measurement on a bulk volume of the sample, such as even after lysing individual components in the bulk volume), count per unit volume of a given component (which is typically measured by identifying such components, but without identifying parameters of individual identified components), and / or concentration of a given component (such as red blood cell concentration, hemoglobin concentration, white blood cell concentration, platelet concentration and / or hematocrit, i.e. red blood cell concentration). Typically, the bulk level of a measured variable is measured by performing a measurement on a bulk volume of the sample. For example, such measurements can include ultrasonic, impedance, optical absorption, transmission, fluorescence, microscopy and / or luminescence measurements performed on a bulk volume of the sample. According to various applications, the first and second measurements are performed on the same portion of the sample, or on different portions of the sample.

[0201] Typically, in a third step 64, a parameter of the sample is determined based on a relationship between the measured variable at the bulk level and the measured variable at the cell level. For some applications, in a sub-step 66 of step 64, the first measurement is normalized with respect to the second measurement. Typically, a relationship between the two measurements is determined (e.g. the two measurements can be divided by each other), and based on the relationship, the concentration or count per unit volume of the reference component is inferred, as described above. For some applications, a second measurement is performed on a second portion of the sample, wherein the second portion of the sample is diluted with respect to the first portion of the sample on which the first measurement is performed, and in sub-step 66, a dilution ratio of the second portion of the sample with respect to the first portion of the sample is determined. For some applications, in another sub-step 68 of step 64, based on the normalization, and other measurements performed on the sample, a parameter of the sample is determined, as described in further detail herein.

[0202] For some applications, the first measurement is performed using a first set of measurement conditions, and the second measurement is performed using a second set of measurement conditions. For some such applications, in a sub-step 70 of step 64, a relationship between the sets of measurement conditions is determined. Typically, in another sub-step 72, based on the relationship between the sets of measurement conditions, a parameter of the sample is determined. For example, first and second optical measurements can be performed on portions of the sample, wherein the sample is disposed in portions of one or more sample chambers having respective dimensions (e.g. respective heights). For some such applications, based on a relationship between the measured variable at the bulk level and the measured variable at the cell level, a relationship between the dimensions of the portions of the one or more sample chambers is determined. Alternatively or in addition, a field of view is determined from which one of the first and second optical measurements is measured (e.g. a microscopic image is acquired), and / or a level of magnification is determined at which one of the first and second optical measurements is measured (e.g. a microscopic image is acquired). For some applications, the measured variable at the bulk level and the measured variable at the cell level are normalized with respect to each other. Subsequently, based on the normalization of the measured variable at the bulk level and the measured variable at the cell level with respect to each other, a parameter of the sample is determined.

[0203] Reference is now made to Figure 4 which is a flowchart showing steps of performing calculations according to some applications of the present application. For some applications, in a first step 80, the hematocrit is measured by performing a first measurement on a blood sample, and in a second step 82, the mean corpuscular volume in the blood sample is measured by performing a second measurement on the blood sample. The hematocrit can be measured, for example, using a micro-hematocrit method, or using ultrasound and / or impedance measurements, and microscopic images obtained from a second portion of the blood sample can be analyzed to measure the mean corpuscular volume. Typically, the second portion of the sample is diluted with respect to the first portion of the blood sample.

[0204] Generally, in the third step 84, a parameter of the sample is determined based on a relationship between hematocrit and mean corpuscular volume. For some applications, in a sub-step 86 of step 84, a first portion of the sample and a second portion of the sample are normalized relative to one another based on a relationship between hematocrit and mean corpuscular volume. Generally, a red blood cell count (e.g., count per unit volume) of the sample is determined by dividing hematocrit by mean corpuscular volume, such that the red blood cell count can thereby serve as a reference parameter against which other parameters can be normalized. For some applications, a count of one or more other components (e.g., red blood cells of a given type, white blood cells, white blood cells of a given type, circulating tumor cells, platelets, platelets of a given type, bacteria, pathogens, pathogens of a given type, reticulocytes, and / or Howell-Jolly bodies) in the sample is determined based on the red blood cell count in the sample. For example, in a sub-step 88 of step 84, a ratio between the red blood cell count and the count of one or more other components in the sample portion can be determined by analyzing microscopic images of the diluted portion of the sample. Subsequently, in a sub-step 89 of step 84, the count of the one or more other components is determined based on the red blood cell count in the source sample portion and the relationship between the red blood cell count and the count of the one or more other components in the diluted portion of the sample.

[0205] Reference is now made to Figure 5 which shows a flowchart of steps implementing an operation in accordance with some applications of the present application. For some applications, in a first step 90, a hemoglobin concentration is measured by performing a first measurement on a blood sample, and in a second step 92, a mean corpuscular hemoglobin in the blood sample is measured by performing a second measurement on the blood sample. For example, the hemoglobin concentration can be measured by performing an optical density measurement on a first portion of the blood sample, and the mean corpuscular hemoglobin can be measured by performing an optical density measurement on a second portion of the sample at the cellular level (i.e., relative to individual cells). Generally, the second portion of the sample is diluted relative to the first portion of the blood sample.

[0206] Generally, in the third step 94, a parameter of the sample is determined based on the relationship between hemoglobin concentration and mean corpuscular hemoglobin. For some applications, in a sub-step 96 of step 94, a first portion of the sample and a second portion of the sample are normalized relative to one another based on the relationship between hemoglobin concentration and mean corpuscular hemoglobin. By dividing the hemoglobin concentration by the mean corpuscular hemoglobin, a red blood cell count (e.g., count / unit volume) in the sample is determined, such that the red blood cell count can thereby serve as a reference parameter relative to which other parameters are normalized. For some applications, based on the red blood cell count in the sample, a count of one or more other components (e.g., red blood cells of a given type, white blood cells, white blood cells of a given type, circulating tumor cells, platelets, platelets of a given type, bacteria, pathogens, pathogens of a given type, reticulocytes, and / or Howell-Jolly bodies) in the sample is determined. For example, in a sub-step 98 of step 94, a ratio between the red blood cell count and the count of one or more other components in a diluted portion of the sample can be determined by analyzing microscopic images of the diluted portion of the sample. Subsequently, in a sub-step 99 of step 94, based on the red blood cell count in the source sample portion and the ratio between the red blood cell count and the count of one or more other components in the diluted portion of the sample, the count of the one or more other components is determined.

[0207] Reference will now be made to Figure 6 which is a flowchart showing steps for performing calculations according to some applications of the present application. For some applications, in a first step 100, the relative amounts of first and second components in a first portion of a blood sample are measured. In a second step 102, a measured variable in a second portion of the blood sample is measured. In a third step 104, a parameter of the blood sample is determined based on the relationship between the relative amounts of the first and second components in the first portion of the blood sample and the measured variable in the second portion of the blood sample. For some applications, the steps described in the flowchart shown in FIG. 1 are performed in conjunction with the steps described in any one of the other flowcharts. Figure 6 the steps described in the flowchart shown in FIG. 1.

[0208] Generally, step 100 is performed by analyzing microscopic images of the first portion of the blood sample. For some applications, the first portion is diluted relative to the second portion, e.g., as described above. (It should be noted that the dilution and source portions of the sample can be described interchangeably with the first and second portions of the sample.)

[0209] For some applications, in step 100, the relative amounts of all white blood cell types, neutrophils, eosinophils, basophils, lymphocytes, monocytes, and / or white blood cell precursors are measured, e.g., by analyzing microscopic images of the first portion of the blood sample. In step 102, the absolute amounts of all types of white blood cells are determined. For some applications, step 102 is performed by performing a bulk level measurement, e.g., by performing an optical density measurement on the source sample portion. In step 104, based on steps 100 and 102, the absolute amounts of various types of white blood cells (or a given type of white blood cell) are determined.

[0210] For some applications, in step 100, the relative amounts of red blood cells, reticulocytes, inclusions, red blood cells with a given morphology, and / or Howell-Jolly bodies are measured, e.g., by analyzing microscopic images of the first portion of the blood sample. In step 102, the absolute amounts of all types of the above components are determined, e.g., by performing an optical density measurement on the source sample portion. In step 104, based on steps 100 and 102, the absolute amounts of various types of the above components (or a given type of the above components) are determined.

[0211] For some applications, in step 100, the relative amounts of reticulated platelets, giant platelets, and / or regular platelets are measured, e.g., by analyzing microscopic images of the first portion of the blood sample. In step 102, the absolute amounts of all platelet types are determined, e.g., by performing an optical density measurement on the source sample portion. In step 104, based on steps 100 and 102, the absolute amounts of various types of platelets (or a given type of platelet) are determined.

[0212] For some applications, a combination of different cell types is analyzed using the techniques described Figure 6 For example, in the first portion, the proportions of any combination of red blood cells, red blood cells of a given type, white blood cells, white blood cells of a given type, platelets, platelets of a given type, inclusions, precursor cells, circulating tumor cells, pathogens, pathogens of a given type, reticulocytes, and / or Howell-Jolly bodies, etc., can be measured, and in the second portion, the absolute counts of any one of the above components can be measured, thereby deriving the absolute counts of another component (modified as appropriate).

[0213] Reference is now made to the following examples, which further illustrate the application. Figure 7which is a flow chart showing steps of performing operations in accordance with some applications of the present application. For some applications, in a first step 110, one or more optical measurement devices are used to perform first and second optical measurements of a sample under different sets of measurement conditions from each other. Typically, in a second step 112, a measured variable of the sample is measured based on the first optical measurement, and in a third step 114, a measured variable of the sample is measured based on the second optical measurement. Depending on the application, the measured variable measured based on the second optical measurement is the same as the measured variable measured based on the first optical measurement, or is different from the measured variable measured based on the first optical measurement. Depending on the application, the first and second optical measurements are performed on the same portion of the sample or on different portions of the sample. For some applications, one optical measurement is performed on a portion of the sample, wherein the portion of the sample is diluted relative to a portion of the sample on which the other optical measurement is performed.

[0214] Typically, in a fourth step 116, a relationship between the measurement conditions of the one or more optical measurement devices used to perform the first and second optical measurements is determined based on a relationship between the measured variable measured based on the first optical measurement and the measured variable measured based on the second optical measurement. For example, a field of view is determined from which one of the first and second optical measurements is measured (e.g., a microscopic image is acquired), and / or a level of magnification is determined at which one of the first and second optical measurements is measured (e.g., a microscopic image is acquired). For some applications, the first and second optical measurements are normalized relative to each other, and a parameter of the sample is determined based on the normalized measurements, e.g., using techniques described herein.

[0215] For some applications, the first measurement is performed using a first type of optical measurement device (e.g., a device configured to perform measurements at the cellular level, e.g., a microscope), and the second measurement is performed using a second type of optical measurement device (e.g., a device configured to perform measurements at the bulk level, e.g., a spectrometer, a luminometer, a spectrophotometer, a camera, a spectral camera, a hyperspectral camera, a fluorometer, a spectrofluorometer, and / or a photodetector). The measurements using the various types of devices are normalized relative to each other to account for errors and / or inaccuracies of one or both of the devices. For example, normalization can account for errors in the magnification level of the microscope, and / or the gain of the device (e.g., a spectrometer, a luminometer, a spectrophotometer, a camera, a spectral camera, a hyperspectral camera, a fluorometer, a spectrofluorometer, and / or a photodetector) configured to perform measurements at the bulk level.

[0216] Reference is now made to Figure 8which is a schematic cross-sectional view of a sample carrier 22 according to some applications of the present application. For some applications, the sample carrier defines one or more sample chambers 120 in which a sample is placed, and the one or more sample chambers define at least a first region 122 (which is shallower) and a second region 124 (which is deeper), the height of the one or more sample chambers changing between the first region and the second region. (For example, as shown, the height of the first region is h, and the height of the second region is (h + Δh)). For some applications, a first optical measurement is performed on a first portion of the sample, wherein the first portion of the sample is disposed in the first region, and a second optical measurement is performed on a second portion of the sample, wherein the second portion of the sample is disposed in the second region. Such measurements can be performed, for example, in accordance with the techniques described in the international application entitled "Sample carrier for optical measurements", filed on even date herewith, which is incorporated herein by reference. For some such applications, the techniques described with reference to Figure 7 step 116 are performed, wherein in step 116, a relationship between the height of the various portions of the one or more sample chambers is determined based on a relationship between the measured variable measured on the basis of the first optical measurement and the measured variable measured on the basis of the second optical measurement.

[0217] Referring to Figures 3-7 the flowchart shown in FIG. 1, it should be noted that the steps of the flowchart need not be performed in the order that they are shown in the flowchart. For some applications, Figures 3-7 the steps of the flowchart shown in FIG. 1 are performed in conjunction with one another. Further, it should be noted that, generally, a response to a sample parameter determined using the techniques described herein results in an output, for example, through the user interface 32 and / or the output device 34, both of which are shown in FIG. 1. Figure 1

[0218] For some applications, the sample described herein is a sample comprising blood or a component thereof (e.g., a diluted or undiluted whole blood sample, a sample comprising primarily red blood cells, or a diluted sample comprising primarily red blood cells), and a parameter related to a component in the blood is determined, for example, a platelet, a white blood cell, an abnormal white blood cell, a circulating tumor cell, a red blood cell, a reticulocyte, a Howell-Jolly body, etc.

[0219] ​Although some applications of the present application have been described with reference to performing a complete blood count and / or with reference to a typical blood analysis, the scope of the present application includes performing other types of analyses using the techniques described herein (modified as appropriate). For example, the techniques described herein can be used in methods related to quantifying blood cells and / or other analytes in blood, in methods for analyzing urine (e.g., for cell clumps), cerebral spinal fluid (CSF), gynecological samples, fecal samples, synovial fluid samples, saliva, semen, sweat, sputum, vaginal secretions, breast milk, bronchoalveolar lavage fluid, gastric lavage fluid, tears, nasal discharge, biological excretions or other biological samples derived from humans or other species. The techniques described are not limited to counting cells, and can be used for quantifying other analytes, such as proteins, peptides, small molecules, infectious agents, etc. The biological samples described can be derived from any living organism, and typically are derived from a homeothermic animal. For some applications, the biological sample is a sample derived from a mammal, such as a sample derived from a human body. For some applications, the sample is taken from any domestic, zoo, and farm animals, including but not limited to dogs, cats, horses, cows, and sheep. Alternatively or additionally, the biological sample is taken from an animal that is a carrier of a disease, including deer or rats.

[0220] For some applications, techniques similar to those described herein are used for non-body samples. For some applications, the sample is an environmental sample, such as a water (e.g., ground water) sample, a surface swab, a soil sample, an air sample, or any combination thereof. In some embodiments, the sample is a food sample, such as a meat sample, a dairy sample, a water sample, a wash sample, a beverage sample, and any combination thereof. For some applications, the techniques described herein are applicable to the analysis of non-biological materials, such as the analysis of analytes in an industrial context.

[0221] Applications described herein can take the form of a computer program product accessible from a computer-usable or computer-readable medium (e.g., a non-transitory computer-readable medium) providing program code for use by or in connection with an instruction execution system (e.g., a computer processor 28). For the purpose of this description, a computer-usable or computer-readable medium can be any apparatus that can contain, store, communicate, propagate, or transport the program code for use by or in connection with the instruction execution system, apparatus, or device. The medium can be electronic, magnetic, optical, electromagnetic, infrared, or semiconductor system (or apparatus or device) or a propagation medium. Typically, the computer-usable or computer-readable medium is a non-transitory computer-usable or computer-readable medium.

[0222] Examples of computer-readable media include semiconductor or solid state memory, magnetic tape, a removable computer diskette, a random access memory (RAM), a read-only memory (ROM), a rigid magnetic disk and an optical disk. Current examples of optical disks include compact disk - read only memory (CD-ROM), compact disk - read / write (CD-R / W) and DVD.

[0223] A data processing system suitable for storing and / or executing program code includes at least one processor (e.g., a computer processor 28) coupled, directly or indirectly, with a system bus to memory elements (e.g., memory 30). The memory elements can include local memory of the processor 28 providing temporary storage of at least some program code to speed the execution process, a removable nonvolatile memory element, or a nonremovable nonvolatile memory element into which system software and / or application programs may be loaded. The data processing system can also read the instructions of the present application from another computer readable medium, such as a storage device or a computer

[0224] A network adapter can also be coupled to the data processing system to enable the data processing system to become coupled to other data processing systems or remote printers or storage devices through intervening private or public networks. Modems, cable modems, and Ethernet cards are just a few of the currently available types of network adapters.

[0225] The computer program code for carrying out operations of the present application can be written in any combination of one or more programming languages, including an object oriented programming language such as Java, Smalltalk, C++ or the like and conventional procedural programming languages, such as the "C" programming language or similar programming languages.

[0226] It should be understood that Figure 3The blocks in the flowcharts shown in 4, 5, 6, and 7, as well as combinations of blocks in the flowcharts, can be implemented by computer program instructions. These computer program instructions can be provided to the processor of a general-purpose computer, a special-purpose computer, or other programmable data processing apparatus to produce a machine such that the instructions, executed by the computer processor (e.g., computer processor 28) or other programmable data processing apparatus, create means for implementing the functions / actions described in detail in the flowcharts and / or the algorithms described in this invention. These computer program instructions can also be stored in a computer-readable medium (e.g., a non-transitory computer-readable medium) that can instruct a computer or other programmable data processing apparatus to function in a particular manner, such that the instructions stored in the computer-readable medium produce an article of manufacture, including means of instruction for implementing the functions / actions described in detail in the flowcharts and algorithms. The computer program instructions can also be loaded into a computer or other programmable data processing apparatus to produce a series of operational steps that execute on the computer or other programmable apparatus, thereby generating a computer-implemented process such that the instructions that execute on the computer or other programmable apparatus provide a process for implementing the functions / actions described in detail in the flowcharts and / or algorithms described in this application.

[0227] Computer processor 28 is typically a hardware device programmed using computer program instructions to create a computer for a specific purpose. For example, when computer processor 28 is programmed to implement reference... Figure 3 When using the algorithms described in 4, 5, 6, and 7, the computer processor typically serves as a special-purpose sample analyzer. Generally, the operations performed by the computer processor 28 according to the present invention convert the physical state of the memory 30 (which is a real physical artifact) into different magnetic poles, charges, etc., depending on the technology of the memory used.

[0228] The apparatus and method described in this invention can be used in conjunction with the apparatus and method described in any of the following patent applications, all of which are incorporated herein by reference:

[0229] Bachelet's US 2012 / 0169863;

[0230] Greenfield's US 2014 / 0347459;

[0231] Pollak's US 2015 / 0037806;

[0232] Pollak's US 20150316477;

[0233] Pollak's US 20160208306;

[0234] US 20160246046 to Yorav Raphael;

[0235] US 20160279633 to Bachelet;

[0236] WO 16 / 030897 to Yorav Raphael;

[0237] WO 17 / 046799 to Eshel; and

[0238] International Application PCT / IL2017 / 050363 to Eshel.

[0239] According to some applications of the present application, the following concepts of the present application are provided:

[0240] 1. A method for a biological sample, the method comprising:

[0241] - measuring a bulk level of a measured variable of the sample by performing a first measurement on the sample;

[0242] - measuring a cellular level of the measured variable of the sample by performing a second measurement on the sample; and

[0243] - determining a parameter of the sample based on a relationship between the bulk level of the measured variable and the cellular level of the measured variable.

[0244] 2. The method of concept 1 of the present application, wherein measuring a parameter of the blood sample comprises normalizing the first and second measurements relative to each other based on a relationship between the bulk level of the measured variable and the cellular level of the measured variable.

[0245] 3. The method of concept 1 of the present application, wherein measuring the bulk level of the measured variable comprises determining an optical density of a given component in the sample.

[0246] 4. The method of concept 1 of the present application, wherein measuring the cellular level of the measured variable comprises analyzing a microscopic image of the sample.

[0247] 5. The method of concept 1 of the present application, wherein performing the first measurement on the sample comprises performing the first measurement on the sample using a first set of measurement conditions, wherein performing the second measurement on the sample comprises performing the second measurement on the sample using a second set of measurement conditions, and wherein determining a parameter of the sample comprises determining a relationship between the measurement conditions used to perform the first and second measurements based on a relationship between the bulk level of the measured variable and the cellular level of the measured variable.

[0248] 6. The method according to concept 1, wherein performing the first measurement comprises performing the first measurement on a first portion of the sample, and wherein performing the second measurement comprises performing the second measurement on the first portion of the sample.

[0249] 7. The method according to any one of concepts 1-5, wherein performing the first measurement comprises performing the first measurement on a first portion of the sample, and wherein performing the second measurement comprises performing the second measurement on a second portion of the sample, wherein the second portion of the sample is different from the first portion of the sample.

[0250] 8. The method according to concept 7, wherein determining the parameter of the sample comprises determining a relationship between the first portion of the sample and the second portion of the sample based on a relationship between the measured variable at the bulk level and the measured variable at the cellular level.

[0251] 9. The method according to concept 7, wherein determining the second portion of the sample to perform the second measurement comprises determining the second portion of the sample to perform the second measurement, wherein the second portion of the sample is diluted relative to the first portion of the sample.

[0252] 10. The method according to concept 9, wherein determining the parameter of the sample comprises determining a normalization factor by determining a property of the first portion of the sample as a parameter that can correct the measurement of the second portion.

[0253] 11. The method according to concept 9, wherein determining the parameter of the sample comprises determining a dilution ratio by which the second portion of the sample is diluted relative to the first portion of the sample.

[0254] 12. The method according to any one of concepts 1-6, wherein the biological sample comprises a blood sample, and wherein determining the parameter of the sample comprises determining a parameter of the blood sample.

[0255] 13. The method according to concept 12, wherein:

[0256] - measuring the measured variable at the bulk level of the sample comprises measuring the hematocrit of the blood sample;

[0257] - measuring the measured variable at the cellular level of the sample comprises measuring the mean corpuscular volume of the blood sample; and

[0258] - determining a parameter of the sample comprises determining a parameter of the sample based on a relationship between the hematocrit and the mean corpuscular volume.

[0259] 14. The method according to concept 12 of the present application, wherein:

[0260] - measuring a bulk level measured variable of the sample comprises measuring a hemoglobin concentration in at least a portion of the blood sample;

[0261] - measuring a cellular level measured variable of the sample comprises measuring a mean corpuscular volume of the blood sample; and

[0262] - determining a parameter of the sample comprises determining a parameter of the sample based on a relationship between the hemoglobin concentration and the mean corpuscular hemoglobin.

[0263] 15. An apparatus for a biological sample, the apparatus comprising:

[0264] - at least one computer processor configured to:

[0265] • measure a bulk level measured variable of the sample by performing a first measurement on the sample;

[0266] • measure a cellular level measured variable of the sample by performing a second measurement on the blood sample; and

[0267] • determine a parameter of the sample based on a relationship between the bulk level measured variable and the cellular level measured variable.

[0268] 16. A computer software product for a biological sample, the computer software product comprising a non-transitory computer readable medium storing program instructions which, when read by a computer, cause the computer to perform the steps of:

[0269] - measuring a bulk level measured variable of the sample by performing a first measurement on the sample;

[0270] - measuring a cellular level measured variable of the sample by performing a first measurement on the sample; and

[0271] - determining a parameter of the sample based on a relationship between the bulk level measured variable and the cellular level measured variable.

[0272] 17. A method for a biological sample, the method comprising:

[0273] - performing first and second optical measurements on the sample using one or more optical measurement devices under measurement conditions that differ from each other for each group;

[0274] - measuring the measured variable of the sample based on the first optical measurement;

[0275] - measuring the measured variable of the sample based on the second optical measurement; and

[0276] - determining the parameter of the sample based on a relationship between the measured variable at the batch level and the measured variable at the cell level.

[0277] - determining a relationship between measurement conditions of one or more optical measurements based on a relationship between the measured variable measured based on the first optical measurement and the measured variable measured based on the second optical measurement, wherein the measurement devices are used for the first and second optical measurements.

[0278] 18. The method according to concept 17, wherein the biological sample comprises a blood sample, and wherein performing first and second optical measurements on the sample comprises performing first and second optical measurements on the blood sample.

[0279] 19. The method according to concept 17, wherein:

[0280] - performing first and second optical measurements on the sample comprises performing first and second optical measurements on portions of the sample, wherein the sample is disposed in portions of one or more sample chambers having various dimensions; and

[0281] - determining a relationship between measurement conditions of the one or more optical measurement devices comprises determining a relationship between the dimensions of the portions of the one or more sample chambers, wherein the optical measurement devices are used to perform the first and second optical measurements.

[0282] 20. The method according to concept 17, wherein performing first and second optical measurements on the sample comprises performing at least one of the first and second optical measurements by acquiring an image of at least a portion of the sample, and wherein determining a relationship between measurement conditions of the one or more optical measurement devices comprises determining a field of view of the image, wherein the measurement devices are used to perform the first and second optical measurements.

[0283] 21. The method according to concept 17, wherein performing the first and second optical measurements on the sample comprises performing at least one of the first and second optical measurements by taking an image of at least a portion of the sample, and wherein determining the relationship between the measurement conditions of the one or more optical measurement devices used to perform the first and second optical measurements comprises determining a magnification level of the image.

[0284] 22. The method according to concept 17, wherein:

[0285] - measuring the measured variable of the sample based on the first optical measurement comprises measuring a given measured variable of the sample based on the first optical measurement;

[0286] - measuring the measured variable of the sample based on the second optical measurement comprises measuring the same given measured variable of the sample based on the second optical measurement; and

[0287] - determining the relationship between the measurement conditions of the one or more optical measurement devices used to perform the first and second optical measurements comprises determining the relationship between the measurement conditions of the one or more optical measurement devices used to perform the first and second optical measurements based on a relationship between a given measured variable measured based on the first optical measurement and a given measured variable measured based on the second optical measurement.

[0288] 23. The method according to concept 17, wherein performing the first optical measurement comprises performing the first optical measurement using a given optical measurement device, and performing the second optical measurement comprises performing the second optical measurement using the same given optical measurement device.

[0289] 24. The method according to any one of concepts 17-22, wherein performing the first optical measurement comprises performing the first optical measurement using a first optical measurement device, and performing the second optical measurement comprises performing the second optical measurement using a second optical measurement device different from the first relationship measurement device.

[0290] 25. The method according to concept 24, wherein:

[0291] - performing the first optical measurement comprises performing the first optical measurement using a first optical measurement device configured to measure a parameter of one or more components in the sample selected from the group consisting of: optical absorption, transmission, fluorescence, and luminescence; and

[0292] - performing the second optical measurement comprises performing the second optical measurement using a microscope configured for acquiring a microscopic image of the sample.

[0293] 26. The method according to any one of the concepts 17-21 or 23 of the present application, wherein:

[0294] - measuring the measured variable of the sample based on the first optical measurement comprises measuring a first measured variable of the sample based on the first optical measurement; and

[0295] - measuring the measured variable of the sample based on the second optical measurement comprises measuring a second measured variable of the sample different from the first measured variable based on the second optical measurement; and

[0296] - determining the relationship between the measurement conditions of the one or more optical measurement devices used for performing the first and second optical measurements comprises determining the relationship between the measurement conditions of the one or more optical measurement devices used for performing the first and second optical measurements based on the relationship between the first and second measured variables.

[0297] 27. The method according to concept 26 of the present application, wherein measuring the first measured variable comprises measuring a batch level measured variable of the sample, and measuring the second measured variable comprises measuring a cell level measured variable of the sample.

[0298] 28. An apparatus for a biological sample, the apparatus comprising:

[0299] - at least one computer processor configured for:

[0300] • performing first and second optical measurements of the sample using one or more optical measurement devices under each set of mutually different measurement conditions;

[0301] • measuring a measured variable of the sample based on the first optical measurement;

[0302] • measuring a measured variable of the sample based on the second optical measurement; and

[0303] • determining the relationship between the measurement conditions of the one or more optical measurement devices used for performing the first and second optical measurements based on the relationship between the measured variable measured based on the first optical measurement and the measured variable measured based on the second optical measurement.

[0304] 29. A computer software product for a biological sample, the computer software product comprising a non-transitory computer readable medium storing program instructions which, when read by a computer, cause the computer to perform the steps of:

[0305] - performing first and second optical measurements on the sample using one or more optical measurement devices under respective sets of different measurement conditions from each other;

[0306] - measuring a measured variable of the sample based on the first optical measurement;

[0307] - measuring a measured variable of the sample based on the second optical measurement; and

[0308] - determining a relationship between the measurement conditions of the one or more optical measurement devices used to perform the first and second optical measurements based on a relationship between the measured variable measured based on the first optical measurement and the measured variable measured based on the second optical measurement.

[0309] 30. A method for a biological sample, the method comprising:

[0310] - performing first and second optical measurements on the sample using one or more optical measurement devices under respective sets of different measurement conditions from each other;

[0311] - measuring a measured variable of the sample based on the first optical measurement;

[0312] - measuring a measured variable of the sample based on the second optical measurement;

[0313] - normalizing the measured variable measured based on the first optical measurement and the measured variable measured based on the second optical measurement with respect to each other; and

[0314] - determining a parameter of the sample based on at least one of the normalized measured variable measured based on the first optical measurement and the normalized measured variable measured based on the second optical measurement.

[0315] 31. An apparatus for a biological sample, the apparatus comprising:

[0316] - at least one computer processor configured to:

[0317] • perform first and second optical measurements on the sample using one or more optical measurement devices under respective sets of different measurement conditions from each other;

[0318] • measure a measured variable of the sample based on the first optical measurement;

[0319] • measuring a measured variable of the sample on the basis of the second optical measurement;

[0320] • normalizing the measured variable measured on the basis of the first optical measurement and the measured variable measured on the basis of the second optical measurement with respect to each other; and

[0321] • determining a parameter of the sample on the basis of at least one of the normalized measured variable measured on the basis of the first optical measurement and the normalized measured variable measured on the basis of the second optical measurement.

[0322] 32. A computer software product for a biological sample, the computer software product comprising a non-transitory computer readable medium storing program instructions which, when read by a computer, cause the computer to carry out the steps of:

[0323] - carrying out first and second optical measurements of the sample using one or more optical measurement devices under each set of mutually different measurement conditions;

[0324] - measuring a measured variable of the sample on the basis of the first optical measurement;

[0325] - measuring a measured variable of the sample on the basis of the second optical measurement

[0326] - normalizing the measured variable measured on the basis of the first optical measurement and the measured variable measured on the basis of the second optical measurement with respect to each other; and

[0327] - determining a parameter of the sample on the basis of at least one of the normalized measured variable measured on the basis of the first optical measurement and the normalized measured variable measured on the basis of the second optical measurement.

[0328] Those skilled in the art will understand that the application is not limited to the particular examples and embodiments described above. Moreover, the scope of the application includes combinations and sub-combinations of the various features described above, as well as variants and modifications thereof which are not prior art and which can be realized by those skilled in the art upon reading the above description.

Claims

1. A method for a blood sample, the method comprising: measuring a hematocrit of a portion of the blood sample by performing a first measurement on a first portion of the blood sample; measuring a mean corpuscular volume of the blood sample by performing a second measurement on a second portion of the blood sample, the second portion being diluted relative to the first portion; and determining a parameter of the blood sample based on a relationship between the hematocrit measured in the first portion and the mean corpuscular volume measured in the second portion by determining a normalization factor defining a property of the first portion of the sample for use as a reference to which other measurements within the sample can be correlated; wherein determining the normalization factor comprises determining a red blood cell count of the sample by dividing the hematocrit by the mean corpuscular volume; and wherein determining a parameter of the blood sample comprises determining a count of one or more components in the sample based on the red blood cell count in the sample by: determining a ratio between the red blood cell count and the count of the one or more components in the portion of the sample by analyzing a microscopic image of the portion of the sample; and and determining the count of the one or more components based on the red blood cell count of the sample and the ratio between the red blood cell count and the count of the one or more components in the portion of the sample.

2. The method of claim 1, wherein performing the first measurement on the blood sample comprises performing a measurement on the blood sample selected from the group consisting of: an ultrasonic measurement and an impedance measurement.

3. The method of claim 1, wherein determining the count of the one or more components based on the red blood cell count of the sample and the ratio between the red blood cell count and the count of the one or more components in the portion of the sample comprises determining a count of a blood component selected from the group consisting of: white blood cells, circulating tumor cells, platelets, pathogens, reticulocytes, and Howell-Jolly bodies.

4. The method of claim 1, wherein determining the count of the one or more components based on the red blood cell count of the sample and the ratio between the red blood cell count and the count of the one or more components in the portion of the sample comprises determining a count of a blood component selected from the group consisting of: red blood cells of a given type, white blood cells of a given type, platelets of a given type, and pathogens of a given type.

5. The method of claim 1, wherein determining a parameter of the sample further comprises determining a concentration of a blood component selected from the group consisting of: hemoglobin, red blood cells, white blood cells, circulating tumor cells, platelets, pathogens, reticulocytes, and Howell-Jolly bodies.

6. The method of claim 1, wherein determining a parameter of the sample further comprises determining a concentration of a blood component selected from the group consisting of: red blood cells of a given type, white blood cells of a given type, platelets of a given type, and pathogens of a given type.

7. The method of claim 1, wherein performing a second measurement on the second portion of the blood sample comprises performing the second measurement by analyzing a microscopic image of the second portion of the blood sample.

8. An apparatus for a blood sample, the apparatus comprising: at least one computer processor configured to: measure a hematocrit of a first portion of the blood sample by performing a first measurement on the first portion of the blood sample; measure a mean corpuscular volume of the blood sample by performing a second measurement on a second portion of the blood sample, the second portion being diluted relative to the first portion; and determine a parameter of the blood sample by determining a normalization factor defining a property of the first portion of the sample for use as a reference for other measurements within the sample that can be correlated based on a relationship between the hematocrit measured within the first portion and the mean corpuscular volume measured within the second portion; wherein determining the normalization factor comprises determining a red blood cell count of the sample by dividing the hematocrit by the mean corpuscular volume; and wherein determining a parameter of the blood sample comprises determining a count of one or more components in the sample based on the red blood cell count in the sample by: determining a ratio between the red blood cell count and the count of one or more components in the portion of the sample by analyzing a microscopic image of the portion of the sample; and determining the count of the one or more components based on the red blood cell count of the sample and the ratio between the red blood cell count and the count of one or more components in the portion of the sample.

9. The apparatus of claim 8, wherein the computer processor is configured to perform the first measurement on the blood sample by performing a measurement selected from the group consisting of: an ultrasonic measurement and an impedance measurement.

10. The apparatus of claim 8, wherein the computer processor is configured to determine the count of the one or more components based on the red blood cell count of the sample and the ratio between the red blood cell count and the count of one or more components in the portion of the sample by determining a count of a blood component selected from the group consisting of: white blood cells, circulating tumor cells, platelets, pathogens, reticulocytes, and Howell-Jolly bodies.

11. The apparatus of claim 8, wherein the computer processor is configured to determine the count of the one or more components based on the red blood cell count of the sample and the ratio between the red blood cell count and the count of one or more components in the portion of the sample by determining a count of a blood component selected from the group consisting of: a given type of red blood cell, a given type of white blood cell, a given type of platelet, and a given type of pathogen.

12. A computer software product for a blood sample, the computer software product comprising a non-transitory computer readable medium storing program instructions which, when read by a computer, cause the computer to perform the steps of: ​ measuring a hematocrit of the first portion of the blood sample by performing a first measurement on the first portion of the blood sample; measuring a mean corpuscular volume of the blood sample by performing a second measurement on a second portion of the blood sample, the second portion being diluted relative to the first portion; and determining a parameter of the blood sample based on a relationship between the hematocrit measured in the first portion and the mean corpuscular volume measured in the second portion by determining a normalization factor defining a property of the first portion of the sample for use as a reference against which other measurements within the sample can be correlated; wherein determining the normalization factor includes determining a red blood cell count of the sample by dividing the hematocrit by the mean corpuscular volume; and wherein determining a parameter of the blood sample includes determining a count of one or more components in the sample based on the red blood cell count in the sample by: determining a ratio between the red blood cell count and the count of one or more components in the portion of the sample by analyzing microscopic images of the portion of the sample; and and determining the count of the one or more components based on the red blood cell count of the sample and the ratio between the red blood cell count and the count of one or more components in the portion of the sample.

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