Differential distribution method

Abstract

The present invention relates to a method for diluting a blood sample for analysis and an apparatus for implementing the method. In the method, an aliquoting device is used, such that a single collection can be performed, a first diluent is formed in a chamber, a portion of the first diluent is collected to form a second diluent in another chamber, blood cells in the first and second chambers are counted, differentiation is performed based on the first diluent, the first chamber is rinsed, a third diluent is formed according to a certain amount of the first diluent liquid retained in the aliquoting device, and then reticulocyte differentiation is performed according to the third diluent.

Description

[0001] The present invention relates to a method for diluting a sample for analysis and a hematology device for implementing such a method. The sample may be blood or other biological fluids, such as puncture fluids, for example cerebrospinal fluid (CSF) containing white blood cells or red blood cells.

[0002] Typically, a hematology device can count and characterize different types of cells present in the blood.

[0003] The known document US 7,661,326 (Beckman Coulter) describes a hematology device that includes a dispensing valve for dividing and dispensing multiple volumes of blood into more than two chambers. Such a device requires a large amount of blood to be collected for simultaneous dispensing into multiple chambers. Therefore, the jet network at the inlet and outlet of the sampling valve is complex, and there is a risk of blockage in the sampling valve due to the circulation of whole blood in the dispensing valve.

[0004] The known document US 6,333,197 (ABX) also describes a needle for collecting blood and injecting it as a reagent into different chambers simultaneously to produce a uniform dilution. The system described in document US 6,333,197 requires a large amount of whole blood to be collected, which is not fully utilized in its design. In addition, since the arrival of the needle and the reagent needs to be aligned, the positioning of the needle in different chambers is complex. Finally, these chambers are specifically designed to homogenize the blood and the reagent.

[0005] The object of the present invention is a new dispensing method that is fast and easy to implement.

[0006] Another object of the present invention is a new method for characterizing blood cells, such as white blood cells and reticulocytes, using a small amount of whole blood.

[0007] At least one of the above objects is achieved by a method for diluting a blood sample for analysis, the method comprising the following steps:

[0008] a) performing a single collection of the sample by an aliquoting device,

[0009] b) injecting the sample into at least one chamber,

[0010] c) diluting the sample in the chamber (referred to as the first chamber if there are multiple chambers) with a dilution reagent to form a first dilution liquid,

[0011] d) collecting a portion of the first dilution liquid by the aliquoting device,

[0012] e) Perform at least two other dilutions to obtain a second diluted liquid and a third diluted liquid, each of the first diluted liquid and the second diluted liquid being directly obtained from the first diluted liquid contained in the aliquoting device, and

[0013] f) During steps a) to e), perform at least one analysis on the first diluted liquid and / or the second diluted liquid and / or the third diluted liquid.

[0014] In the method according to the invention, with a single sample collection, three dilutions are performed, making it possible to perform a complete analysis of the sample. This single collection can be, for example, an amount of 20 μl of whole blood or diluted blood, while in the prior art, this collection is typically about 120 μl or more. Because the first diluted liquid is cleverly retained in the aliquoting device for the second and third dilutions, this method has the advantage of being simple to implement.

[0015] In addition, the method according to the invention allows for a high analysis speed.

[0016] It should be understood that the second diluted liquid and the third diluted liquid are obtained independently of each other, i.e., the third diluted liquid is not obtained from the second diluted liquid, but directly from the first diluted liquid retained in the aliquoting device. The dilution can be carried out continuously in a single chamber or multiple chambers.

[0017] Depending on the configuration selected with one chamber or more chambers, all or part of the analysis can be carried out continuously or simultaneously (in parallel).

[0018] According to one embodiment, the analysis can include characterizing the first diluted liquid and / or the second diluted liquid and / or the third diluted liquid by optical measurement for counting and / or differentiating the particles contained in the liquid.

[0019] According to one embodiment, the analysis can include counting the particles in the first diluted liquid and / or the second diluted liquid and / or the third diluted liquid by a resistance sensor.

[0020] The optical measurement can be carried out on an optical bench or in the chamber for dilution, which is equipped with optical means.

[0021] According to the invention, one or more resistance sensors can be connected or incorporated into at least one chamber or optical bench.

[0022] "Optical bench" means a device capable of performing the following operations:

[0023] - Counting particles by optical means,

[0024] - Characterizing particles by optical means,

[0025] - Counting particles by incorporating one or more resistance sensors therein, and

[0026] - Characterizing the particles by an optical device enhanced with information from a resistance device.

[0027] According to an embodiment of the present invention, step e) can include the following steps:

[0028] e1) Injecting a first quantity of a first dilution liquid contained in an aliquoting device into a chamber, preferably into a second chamber, and retaining a second quantity of the first dilution liquid in the aliquoting device,

[0029] e2) Diluting the first dilution liquid contained in the second chamber with a dilution reagent to constitute a second dilution liquid,

[0030] e3) Injecting a lysis solution into the first chamber to lyse red blood cells,

[0031] e4) After transferring a portion of the first dilution liquid to an optical bench, differentiating white blood cells in the first dilution liquid contained in the first chamber by optical measurement directly in the first chamber or on the optical bench,

[0032] e5) Counting red blood cells and / or platelets in the second dilution liquid, preferably in the second chamber, but this can also be done on the optical bench,

[0033] e6) Counting white blood cells and / or measuring hemoglobin in the first dilution liquid, preferably in the first chamber, but this can also be done on the optical bench,

[0034] e7) Flushing at least one chamber,

[0035] e8) Injecting a portion of the second quantity of the first dilution liquid still contained in the aliquoting device into the flushed chamber,

[0036] e9) Diluting the liquid contained in the flushed chamber with a dilution reagent to constitute a third dilution liquid,

[0037] e10) Analyzing the third dilution liquid.

[0038] One set of counting and / or differentiation measurements can be performed on at least one chamber (preferably two chambers) and an optical bench with only one sample collection.

[0039] Using such a method, the analysis speed is very fast. For example, at least 60 tests can be performed per hour, and one test includes counting red blood cells, counting white blood cells, and differentiating white blood cells.

[0040] According to an advantageous feature of the present invention, in step e10), a portion of the third dilution liquid can be transferred to the optical workbench for differentiating red blood cells, particularly immature red blood cells, reticulocytes.

[0041] Using the method according to the present invention, a single sample collection allows for the differentiation of white blood cells and the differentiation of red blood cells. In particular, for example, two counts and two differentiations can be performed with a single sample collection of 20 μl, one or two chambers, and a single optical workbench.

[0042] According to an advantageous feature of the present invention, an aliquoting device can be used, comprising:

[0043] - a needle capable of moving between a sample collection area and the at least one chamber,

[0044] - a dilution reagent dispenser, and

[0045] - a sampling valve, which includes at least two liquid passages and a calibration volume channel, wherein a first liquid passage connects the dispenser to the needle, a second liquid passage connects the dispenser to a second chamber, and the calibration volume channel activates the first liquid passage or the second liquid passage.

[0046] The sampling valve can be designed to include two ceramic discs, one of which contains the calibration volume channel. This channel can be shifted between two positions, a first position where the channel is contained within the first liquid passage and a second position where the channel is contained within the second liquid passage.

[0047] The present invention is particularly significant for the reuse of the first dilution liquid present in the needle up to the sampling valve.

[0048] In prior art systems, there may be a risk of valve blockage due to the circulation of whole blood in the fluid channels of the valve. In the present invention, since it is diluted blood that circulates in the channels of the valve, this risk is quite limited.

[0049] Step d) can preferably be carried out by collecting the sample and retaining it within the needle and the calibration volume channel of the sampling valve. In order to inject the first dilution liquid into the chamber for the second dilution, the calibration volume channel containing the first quantity of the first dilution liquid can be moved to the second liquid passage and then injected into the first chamber or the second chamber through the second liquid passage. In the calibration volume channel of the sampling valve, the first quantity is precisely calibrated.

[0050] The sampling valve forms part of the fluid circuit for collecting the first diluent.

[0051] This makes it possible to increase the speed and avoid contaminating the needle. In an embodiment, the needle only performs a single collection of the starting sample and is subsequently only used to collect the first dilution liquid. In fact, the needle makes it possible to collect the first diluent in the first chamber, but the injection into the second chamber is carried out directly through the second liquid passage. More precisely, the tube can connect the sampling valve to the second chamber. This feature makes it possible, for example, to perform a second dilution while retaining a portion of the first diluent in the needle, which makes it possible to subsequently dispense a portion of the first diluent into the first chamber for a third dilution without having to perform a new sample collection.

[0052] According to an embodiment of the invention, step d) can be carried out by collecting the first dilution liquid and retaining it within the needle and the sampling valve. The first dilution liquid is preferably aspirated into the sampling valve and passes over the valve into the tube between the sampling valve and the dispenser. This embodiment ensures the complete filling of the calibration volume channel, since the volume of the calibration volume channel is precisely calibrated, thus allowing an accurate second dilution; thus, the volume injected into the second chamber is precisely known.

[0053] According to another embodiment of the invention, an aliquoting device comprising one or more sets of precision pistons / syringes can be used in order to collect samples and different diluents from different chambers and inject the samples and different diluents into different chambers. The injection volumes for the first dilution and the third dilution are determined by precisely controlling the pistons / syringes.

[0054] The steps of the first dilution and the second dilution can preferably be carried out by injecting a dilution reagent by means of the aliquoting device. When the aliquoting device comprises a needle and a sampling valve, the two liquid passages comprise a tube in which the dilution reagent from the dilution reagent dispenser serves as the liquid for dispensing the sample and / or the dilution reagent.

[0055] According to a feature of the invention, all or some of the dilution steps are carried out by injecting the dilution reagent into one or more chambers from a liquid passage independent of the aliquoting device.

[0056] Advantageously, a single optical bench connected to the first chamber can be used.

[0057] According to an advantageous feature of the invention, all or part of the counting is carried out by means of a resistance sensor connected to the first chamber and / or the second chamber and / or other chambers (if there are more than two chambers).

[0058] By way of example, the ratio of the first dilution can be 1 / 200, the ratio of the second dilution can be 1 / 10000, and the ratio of the third dilution can be 1 / 10000.

[0059] According to an embodiment of the present invention, the injection of the lysis solution in step e3) can be carried out through a liquid passage independent of the aliquoting device and directly enter the first chamber.

[0060] This lysis solution has the function of destroying red blood cells and separating white blood cells. This also allows hemoglobin to be stabilized in the form of a stable complex.

[0061] According to an advantageous feature of the present invention, the method can include the step of adding a fluorescent dye to the first chamber before each optical differentiation measurement. It is possible to preferably use an optical bench capable of detecting fluorescence. Due to the presence of the fluorescent dye, reticulocytes, i.e., immature red blood cells, can be detected.

[0062] In fact, the fluorescent dye can be added to the first dilution liquid and / or the second dilution liquid and / or the third dilution liquid before any optical measurement in order to improve the differentiation of blood cells (such as white blood cells) or / and the characterization of reticulocytes using fluorescence.

[0063] Advantageously, an independent liquid passage of the aliquoting device can be used, which is directly connected to the chamber for the flushing step.

[0064] It is also possible to use an independent liquid passage of the aliquoting device, which is directly connected to the chamber for dilution step e9). This can be the same liquid passage as for flushing or a different liquid passage.

[0065] According to the present invention, steps e5) and e6) can be carried out in parallel or sequentially.

[0066] Parallel counting is carried out using a single aspiration system, allowing simultaneous aspiration from two chambers into different channels. It is perfectly conceivable to carry out separate (non-simultaneous) counting using a single or multiple different aspiration systems.

[0067] According to another aspect of the present invention, there is provided a hematology device for automatically counting and differentiating cells in a blood sample, characterized in that the hematology device comprises:

[0068] - at least one chamber,

[0069] - at least one optical bench connected to at least one chamber,

[0070] - an aliquoting device, which comprises:

[0071] - a needle capable of moving between a sample collection area and at least one chamber,

[0072] - a dilution reagent dispenser, and

[0073] - Sampling valve, which includes at least two liquid passages and a calibration volume passage, wherein a first liquid passage connects the dispenser to the needle, a second liquid passage connects the dispenser to at least one chamber, and the calibration volume passage activates the first liquid passage or the second liquid passage.

[0074] A processing unit for implementing different steps and controlling different components is also provided.

[0075] The sampling valve according to the present invention can include a calibration volume passage, which can form part of the first liquid passage or part of the second liquid passage. In other words, the calibration volume passage switches from one liquid passage to another. When on the first liquid passage, the dispenser can control the suction or discharge of a part of the liquid contained in the first liquid passage, and the second liquid passage is inoperable. When on the second liquid passage, the dispenser can control the discharge of a part of the liquid contained in the second liquid passage, and the first liquid passage is inoperable.

[0076] By studying the detailed description and the drawings of non-limiting embodiments, other advantages and features of the present invention will become apparent, in the drawings:

[0077] Figure 1 is a schematic diagram showing several components constituting an available automatic hematology analyzer,

[0078] Figure 2 is a schematic diagram showing the preliminary steps of whole blood collection,

[0079] Figure 3 is a schematic diagram showing step 1 of constituting the first dilution,

[0080] Figure 4 is a schematic diagram showing step 2 of collecting a part of the first dilution,

[0081] Figure 5 is a schematic diagram showing step 3 of constituting the second dilution,

[0082] Figure 6 is a schematic diagram showing step 4 of transferring to the optical workbench for white blood cell differentiation,

[0083] Figure 7 is a schematic diagram showing step 5 of emptying and flushing the chamber,

[0084] Figure 8 is a schematic diagram showing step 7 of constituting the third dilution,

[0085] Figure 9 is a schematic diagram showing step 8 of transferring to the optical workbench for red blood cell differentiation,

[0086] Figure 10 It is a schematic diagram showing step 9 of emptying and final rinsing.

[0087] The embodiments to be described below are in no way restrictive; specifically, if the selection of features is sufficient to confer a technical advantage or to distinguish the present invention from the prior art, a variant of the present invention can be implemented that includes only the following selection of features separated from the other features. This selection includes at least one feature that is preferably functional, has no structural details, or only has a part of the structural details, if that part alone is sufficient to confer a technical advantage or to distinguish the present invention from the state of the prior art.

[0088] Specifically, all the variants and all the embodiments described are provided to be combined with each other in any combination that is not objected to from a technical point of view.

[0089] In the drawings, elements common to multiple drawings retain the same reference numerals.

[0090] Figure 1 It shows the components that make up an automated hematology analyzer, which is ready and waiting for the analysis cycle.

[0091] The optical bench 1 for characterizing different types of cells present in the blood can be seen. The first chamber 2 is connected to the optical bench 1 through a solenoid valve 3, which can block or allow the fluid contained in the first chamber 2 to flow to the optical bench 1. The first chamber 2 includes an outlet 21 connected to the solenoid 3, and electronic devices for resistivity measurement, in particular at least one sensor 22. These measurements are carried out, for example, during cell counting.

[0092] For the sake of clarity of illustration, only the optical bench 1 is shown; it is obvious that a flow cell (not shown) is provided in this optical bench, and the fluid to be characterized can flow in this flow cell.

[0093] The dilution reagent dispenser 4 can also be seen, which is connected to the sampling valve 5 through two parallel conduits C1 and C2. One side of the sampling valve 5 is connected to the needle 6 through the conduit C3, and the other side is connected to the second chamber 7 through the conduit C4.

[0094] The sampling valve 5 is a valve including two liquid passages and a calibration volume passage 8. The first liquid passage enables the connection of the catheter C1 and C3 through the calibration volume passage 8. The second liquid passage enables the connection of the catheter C2 and C4 through the calibration volume passage 8. Thus, the calibration volume passage can form part of the first liquid passage or part of the second liquid passage, but not both simultaneously. Advantageously, the passage 8 of the calibration volume is a catheter adapted to switch from one liquid passage to another and forms a reservoir for the fluid, the volume of which is very precisely predetermined. Thus, a predetermined amount of liquid can be transported from one liquid passage to another.

[0095] The catheter C4 is connected to the second chamber 7 through the inlet 71.

[0096] The second chamber 7 also includes electronic devices for resistivity measurement, in particular at least one sensor 72. These measurements are carried out, for example, during cell counting. An independent liquid passage 74 can also be provided for injecting dilution reagents.

[0097] It can also be seen a processing unit 9 capable of controlling different components.

[0098] In Figure 1 , the needle 6, the first liquid passage and the second liquid passage, the calibration volume passage 8 and the chamber are filled with a clean dilution reagent. The machine can be used.

[0099] In Figure 2 In the preparatory step, blood is collected from the whole blood vessel 10 into the needle 6. Then, a certain amount of blood is only located in a part of the needle. Except for the part of the needle 6 containing blood, the first liquid passage including the catheters C1 and C3 is mainly filled with dilution reagent. The needle collects blood through the suction function of the dilution reagent dispenser.

[0100] At the same time, the first chamber 2 is emptied.

[0101] In step 1, in Figure 3 , the needle 6 moves into the first chamber 2 in order to inject all the collected blood into it. The injection continues in order to fill it with the dilution reagent contained in the first liquid passage and transported through the dispenser. The mixture of the blood thus deposited and the dilution reagent, which is much larger in volume than the collected blood, constitutes the first dilution liquid, the ratio of which is, for example, one volume of blood to two hundred volumes of dilution reagent.

[0102] In step 2, in Figure 4 , a part of the first diluent is collected from the first chamber 2 into a part of the catheter C1. Thus, the needle 6, the catheter C3 and the sampling valve, in particular the calibration volume passage 8, are completely filled with the first dilution liquid.

[0103] In step 3, in Figure 5 the calibration volume channel 8 filled with the first dilution liquid is switched from the first liquid path to the second liquid path; the latter has now started operating. The fact that the first dilution liquid has been aspirated into a part of the catheter C1 in step 2 enables the channel 8 of the calibration volume to be completely filled.

[0104] Then, the liquid contained in the second liquid path is pushed so that the amount contained in the calibration volume channel 8 and most of the dilution reagent are injected into the second chamber 7 via the inlet 71 and the catheter C4. Thus, a second dilution liquid is formed having, for example, a ratio of one volume of clean blood to ten thousand volumes of dilution reagent.

[0105] In the first chamber, once the required portion of the first dilution liquid has been collected, the needle 6 is raised again so that it is not in contact with the liquid in the first chamber 2, and the lysis solution is injected into the first chamber 2 through the inlet 23. The lysis solution has the effect of destroying red blood cells.

[0106] At this stage, it should be noted that the first dilution liquid still remains in the needle 6 and in a part of the first liquid path including the catheter C1 and the catheter C3.

[0107] In Figure 6 step 4, the solution is transferred from the first chamber 2 to the optical bench for the differentiation of the white blood cell population. In parallel or separately, the white blood cells are counted in the first chamber 2 by measuring the resistivity, and the hemoglobin measurement is performed by a spectrophotometer (not shown).

[0108] In the second chamber 7, the red blood cells and platelets are counted by measuring the resistivity. The counting in the second chamber 7 can be carried out simultaneously with the counting in the first chamber. This is especially the case when both chambers use a single aspiration system (not shown) during the counting process. In fact, the counting sequence requires aspirating the liquid contained in the chamber through the calibration orifice based on the impedance measurement principle by creating a vacuum.

[0109] In Figure 7 step 5, both chambers are completely rinsed and emptied, and the same applies to the fluid circuit between the first chamber 2 and the optical bench 1. The chamber can be rinsed with the dilution reagent so that the dilution reagent is conveyed to the optical bench in the fluid circuit, thereby rinsing and refilling the circuit. Then, the calibration volume channel 8 filled with the dilution reagent is switched to the first liquid path.

[0110] In Figure 8In step 7, a third dilution is performed according to the present invention. To this end, a certain amount of the first dilution liquid still present in the needle 6 is injected into the first chamber 2. A specific volume is pushed. At the end of this step, a residual first dilution volume can still be present in the needle 6. The dilution is carried out by injecting a dilution reagent via the inlet 24 of the first chamber 2. The supply circuit of this inlet 24 of the dispenser 4 is not shown. A fluorescent dye can also be added.

[0111] In Figure 9 step 8, the solution is transferred from the first chamber 2 to the optical bench 1; then the differentiation of red blood cells and reticulocytes is carried out.

[0112] In Figure 10 step 9, when the differentiation is complete, the needle 6 is emptied of the residual blood. The chamber is rinsed and then refilled with the dilution reagent, awaiting subsequent analysis.

[0113] Therefore, the present invention enables the ingenious use of an aliquoting device to perform multiple differentiation measurements based on a single collection, and this aliquoting device allows the first dilution liquid and the dilution reagent to be segmented.

[0114] Therefore, the present invention relates to a method for diluting a blood sample for analysis and a device for implementing such a method.

[0115] In this method, an aliquoting device is used, enabling a single collection, forming a first dilution in a chamber, collecting a portion of the first dilution to form a second dilution in another chamber, counting blood cells in the first and second chambers, performing differentiation based on the first dilution, rinsing the first chamber, forming a third dilution starting from a certain amount of the first dilution retained in the aliquoting device, and then performing the differentiation of reticulocytes based on this third dilution.

[0116] Of course, the present invention is not limited to the examples just described, and many adjustments can be made to these examples without departing from the scope of the present invention.

Claims

1. A method for diluting a blood sample for analysis, the method comprising the following steps: a) performing a single collection of the sample by an aliquoting device, b) injecting the sample into a first chamber (2), c) diluting the sample in the chamber with a dilution reagent to form a first diluted liquid, d) collecting a portion of the first diluted liquid by the aliquoting device, e) performing at least two other dilutions to obtain a second diluted liquid and a third diluted liquid, each of the second diluted liquid and the third diluted liquid being directly obtained from the first diluted liquid contained in the aliquoting device, and f) during steps a) to e), performing at least one analysis on the first diluted liquid and / or the second diluted liquid and / or the third diluted liquid; characterized in that the aliquoting device further comprises: - a needle (6) capable of moving between a sample collection area and the at least one chamber (2, 7), - a dilution reagent dispenser (4), and - a sampling valve (5) comprising at least two liquid passages and a calibration volume passage (8), wherein a first liquid passage (C1, 8, C3) connects the dispenser (4) to the needle (6), a second liquid passage (C2, 8, C4) connects the dispenser (4) to a second chamber (7), and the calibration volume passage (8) activates the first liquid passage or the second liquid passage; wherein step d) is performed by collecting the first diluted liquid and retaining it within the needle (6) and the calibration volume passage (8) of the sampling valve (5), and wherein, in order to inject the first diluted liquid into a chamber for a second dilution, the calibration volume passage containing a first quantity of the first diluted liquid is switched from the first liquid passage to the second liquid passage and then injected into the second chamber through the second liquid passage, the first quantity being precisely calibrated in the calibration volume passage (8) of the sampling valve (5), and the third diluted liquid being directly taken from the first diluted liquid contained within the needle.

2. The method according to claim 1, wherein The analysis comprises characterizing the first diluted liquid and / or the second diluted liquid and / or the third diluted liquid by optical measurement for counting and / or differentiating the particles contained in the liquid.

3. The method according to claim 1 or 2, characterized in that, The analysis comprises counting the particles of the first diluted liquid and / or the second diluted liquid and / or the third diluted liquid by a resistance sensor.

4. The method according to claim 1 or 2, characterized in that, The analysis comprises characterizing the third diluted liquid by optical measurement for counting and / or differentiating the particles contained in the liquid.

5. The method according to claim 1 or 2, characterized in that, Step e) comprises the following steps: e1) injecting a first quantity of the first diluted liquid contained in the aliquoting device into a second chamber and retaining a second quantity of the first diluted liquid in the aliquoting device, e2) diluting the first diluted liquid contained in the second chamber with a dilution reagent to form a second diluted liquid, e3) injecting a lysis solution into the first chamber to lyse red blood cells, e4) After transferring a portion of the first dilution liquid to the optical workbench, white blood cells in the first dilution liquid contained in the first chamber are differentiated by optical measurement directly in the first chamber or on the optical workbench. e5) Count red blood cells and / or platelets in the second dilution liquid. e6) Count white blood cells in the first dilution liquid and / or measure hemoglobin in the first dilution liquid. e7) Rinse at least one chamber. e8) Inject a portion of the second quantity of the first dilution liquid still contained in the aliquoting device into the rinsed chamber. e9) Dilute the liquid contained in the rinsed chamber with a dilution reagent to form a third dilution liquid. e10) Analyze the third dilution liquid.

6. The method according to claim 5, characterized in that, In step e10), a portion of the third dilution liquid is transferred to the optical workbench for the differentiation of reticulocytes.

7. The method according to claim 1 or 2, characterized in that, The steps of the first dilution and the second dilution are performed by injecting a dilution reagent by the aliquoting device.

8. The method according to claim 1 or 2, characterized in that, All or some of the dilution steps are performed by directly injecting the dilution reagent from a liquid passage (24, 74) independent of the aliquoting device into one or more chambers.

9. The method according to claim 1 or 2, characterized in that, Use a single optical workbench connected to the first chamber.

10. The method according to claim 1 or 2, characterized in that The ratio of the first dilution is 1 / 200, the ratio of the second dilution is 1 / 10000, and the ratio of the third dilution is 1 / 10000.

11. The method according to claim 1 or 2, characterized in that, Use a fluorescence optical workbench for analysis.

12. The method according to claim 5, wherein Count red blood cells and / or platelets in the second dilution liquid in the second chamber or on the optical workbench.

13. The method according to claim 5, wherein Count white blood cells in the first dilution liquid in the first chamber or on the optical workbench and / or measure hemoglobin in the first dilution liquid.

14. A hematology device for automatically counting and differentiating cells in a blood sample, characterized in that, The hematology device comprises: - At least one chamber (2, 7), - At least one optical workbench (1) connected to at least one chamber, - An aliquoting device, which comprises: - A needle (6) capable of moving between a sample collection area and at least one chamber (2, 7), - A dilution reagent dispenser, wherein the aliquoting device further comprises: - A sampling valve, which comprises at least two liquid passages and a calibration volume channel (8), a first liquid passage (C1, 8, C3) connecting the dilution reagent dispenser to the needle, a second liquid passage (C2, 8, C4) connecting the dilution reagent dispenser to at least one chamber, and the calibration volume channel (8) activating the first liquid passage or the second liquid passage, - And a processing unit (9), which is configured to control different components of the device and is also configured to implement different steps of the method according to claim 1.

15. The device according to claim 14, characterized in that, The calibration volume channel (8) contains a calibration volume conduit, and the calibration volume channel can form a part of the first liquid passage or a part of the second liquid passage.

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