Fluid transport in biological analytical systems

Abstract

The biological analysis system includes a probe. The probe is configured to dispense a biological sample into a first chamber. The system also includes a pump. The pump is configured to deliver a first delivery of a diluent to the first chamber for mixing with the biological sample to generate a sample mixture, aspirate a portion of the sample mixture from the first chamber, and deliver a portion of the sample mixture to a second chamber. A method of analyzing a biological sample includes delivering the diluent to the first chamber via the first pump, dispensing a blood sample into the first chamber via the probe to generate a sample mixture, and delivering a portion of the sample mixture from the first chamber to the second chamber via the first pump.

Description

[Technical field]

[0001] Priority This application claims the benefit of U.S. patent application Ser. No. 63 / 292,806, entitled "Fluid Transfer in a Biological Analysis System," filed Dec. 22, 2022, the disclosure of which is incorporated herein by reference. [Background technology]

[0002] Various types of tests related to the diagnosis and treatment of patients can be performed by analyzing patient samples. This can include analyzing the patient's microbes or "microbes," as well as analyzing samples to determine chemistry, antigens, antibodies, blood counts, and other factors that can affect the patient's health. Microbes are microscopic organisms, such as bacteria, fungi, or viruses, which can be unicellular or multicellular. In analyzing microbes, biological samples containing the patient's microbes can be taken from the patient's infectious material, body fluids, or abscesses, placed in a test panel or array, combined with various reagents, cultured, and analyzed to aid in the treatment of the patient. Analysis of the patient's chemistry, immunoassays, blood counts, and other characteristics can be performed as well. For these various analyses, automated biochemical analyzers or biological testing systems have been developed to meet the needs of medical facilities and other institutions to facilitate the analysis of patient samples, improve the accuracy and reliability of results compared to analyses using manual operations, and help determine the effectiveness of various antimicrobial agents.

[0003] Biological samples are typically dispensed into various chambers for analysis, processing, and / or testing within the chambers. For example, a blood analyzer may include a sample probe (also called an aspirating-dispensing probe) that may aspirate a biological sample (e.g., blood) from a specimen and then dispense the biological sample into a first bath (e.g., a white blood cell bath). The probe may then aspirate a portion of the sample solution from the first bath and then dispense the aspirated portion into a second bath (e.g., a red blood cell bath), such that each portion of the sample solution in the first and second baths may be analyzed separately from each other. The probe must travel along a path that includes at least the specimen, the first bath, and the second bath. Such a path may be complex and / or have a relatively large footprint, and the size of the analyzer may have to be undesirably large to accommodate this path. The probe must also sequence at least four separate tasks (e.g., first aspiration, first dispense, second aspiration, and second dispense) to facilitate delivery of sample solution to each bath. Such reliance on the probe to perform all of these tasks may prevent the probe from performing other functions, such as aiding in further analysis (e.g., measurement, imaging, and / or flow imaging) of red blood cells (RBCs), white blood cells (WBCs), and platelets (PLTs), and / or may undesirably limit the throughput of the analyzer. Summary of the Invention

[0004] In some embodiments, a biological analysis system is described that includes a probe. The probe is configured to dispense a biological sample into a first chamber. The system also includes a pump. The pump is configured to deliver a first delivery of a diluent to the first chamber, aspirate a portion of the sample mixture from the first chamber, and deliver a portion of the sample mixture to a second chamber for mixing with the biological sample to generate a sample mixture. In some embodiments, the probe is configured to aspirate the biological sample from the biological specimen prior to dispensing the biological sample into the first chamber. Additionally or alternatively, the pump may include a syringe pump. The system may include a complete blood count (CBC) drip chamber, the CBC drip chamber configured to count blood cells in at least one of the first or second chambers. Additionally or alternatively, the pump may be configured to deliver a second delivery of a diluent to the second chamber while delivering a portion of the sample mixture to the second chamber. The system may include a lysate pump, the lysate pump configured to deliver a lysate to one of the chambers. The system may include a first and a second chamber, the first and second chambers configured to be selectively fluidly coupled to one another. The system may further include a transport line configured to selectively fluidly couple the first and second chambers to one another, and the pump may be configured to aspirate a portion of the sample mixture into the transport line prior to conveying the portion of the sample mixture through the transport line to the second chamber. The system may further include at least one diluent push / pull line configured to selectively fluidly couple to the transport line, and the pump may be configured to draw an air gap in the at least one diluent push / pull line while aspirating the portion of the sample mixture into the transport line.

[0005] In some embodiments, a biological analysis system is described that includes a first chamber, a second chamber, and a probe. The probe is configured to dispense a biological sample into the first chamber. The system also includes a first pump. The first pump is configured to transport a portion of the biological sample from the first chamber to the second chamber. The system also includes a second pump. The second pump is configured to deliver a fluid medium to at least one of the first chamber or the second chamber. In some embodiments, the fluid medium includes a lysing solution, and the second pump is configured to deliver the lysing solution to the first chamber after transporting the portion of the biological sample to the second chamber. The system may further include a CBC drip chamber, the CBC drip chamber being fluidly coupled to at least one of the first or second chamber for counting blood cells in at least one of the first or second chamber. Additionally or alternatively, the first pump may be configured to deliver a first delivery of diluent to the first chamber for mixing with the biological sample to generate a sample mixture prior to transporting the portion of the biological sample to the second chamber. The first pump may be configured to deliver a second delivery of diluent to the second chamber while transporting the portion of the biological sample to the second chamber. In some embodiments, the first and second chambers are configured to be selectively fluidically coupled to each other. The system may further include a transport line configured to selectively fluidically couple the first and second chambers to each other and at least one diluent push / pull line configured to be selectively fluidically coupled to the transport line, and the first pump may be configured to draw an air gap in the at least one diluent push / pull line while aspirating the portion of the biological sample into the transport line. In some embodiments, the first chamber comprises a hemoglobin transducer. The hemoglobin transducer may include a filtered light source and an optical sensor.

[0006] In some embodiments, a method for analyzing a biological sample is described. The method for analyzing a biological sample includes: delivering a diluent to a first chamber via a first pump; dispensing a blood sample into the first chamber via a probe to generate a sample mixture; and delivering a portion of the sample mixture from the first chamber to a second chamber via the first pump. The method may further include delivering a dissolving solution to the first chamber or the second chamber via the second pump.

[0007] While this specification particularly describes and concludes with claims which distinctly claim the invention, it is believed the invention will be better understood from the following description of specific examples taken in conjunction with the accompanying drawings, in which like reference numerals identify like elements and in which: [Brief description of the drawings]

[0008] [Figure 1] 1 is a schematic diagram of an exemplary biological analysis system. [Figure 2A] FIG. 2 is a schematic diagram of the biological analytical system of FIG. 1 , showing the flushing of the WBC and RBC baths of the system via wash and waste pumps, and the use of diluent to cleanse the WBC and RBC baths of the waste fluids contained therein. [Figure 2B] FIG. 2 is a schematic diagram of the biological analytical system of FIG. 1, illustrating the delivery of diluent to the WBC bath via a wash pump. [Figure 2C] FIG. 2 is a schematic diagram of the biological analytical system of FIG. 1, illustrating the delivery of a blood sample to a WBC bath via a probe of the system. [Figure 2D] FIG. 2 is a schematic diagram of the biological analysis system of FIG. 1, illustrating the further delivery of diluent to the WBC bath via a wash pump to generate a prediluent solution in the WBC bath. [Figure 2E] FIG. 2 is a schematic diagram of the biological analysis system of FIG. 1, illustrating the extraction of a portion of the prediluent solution from the WBC bath via a wash pump. [Figure 2F]FIG. 2 is a schematic diagram of the biological analytical system of FIG. 1, illustrating the delivery of a transport portion of the pre-diluent solution to the RBC input line of the system via a wash pump. [Figure 2G] FIG. 2 is a schematic diagram of the biological analytical system of FIG. 1, illustrating the delivery of lysate to the WBC bath via the system's lysate pump. [Figure 2H] FIG. 2 is a schematic diagram of the biological analysis system of FIG. 1, illustrating the further delivery of diluent to the WBC bath via a wash pump to generate the final WBC solution. [Figure 2I] FIG. 2 is a schematic diagram of the biological analytical system of FIG. 1, illustrating the delivery of the transport portions of the diluent and pre-diluent solutions to the RBC bath via a wash pump. [Diagram 3] FIG. 2 illustrates a method of analyzing a biological sample using the biological analysis system of FIG. 1. [Figure 4] FIG. 2 is a schematic diagram of another exemplary biological analysis system. [Figure 5A] 5 is a schematic diagram of the biological analytical system of FIG. 4 showing delivery of a blood sample to the WBC bath of the system via a probe of the system, with diluent having been previously delivered to the WBC bath via a diluent pump of the system, and further showing delivery of mixing bubbles of air to the WBC bath via a mixing pump of the system to generate a pre-diluent solution in the WBC bath, delivery of diluent to a sweep flow tank of the system via a wash pump of the system, and filling of the diluent pump with diluent. [Figure 5B] FIG. 5 is a schematic diagram of the biological analysis system of FIG. 4, illustrating the extraction of a portion of the pre-diluent solution from the WBC bath via a diluent pump. [Figure 5C] FIG. 5 is a schematic diagram of the biological analytical system of FIG. 4 , showing the delivery of transport portions of diluent and pre-diluent solutions to the RBC bath of the system via the diluent pump to produce the final RBC solution, and further showing the delivery of lysing solution to the WBC bath via the lysing solution pump of the system, and the further delivery of mixing bubbles of air to the WBC bath via the mixing pump to produce the final WBC solution. [Figure 5D]FIG. 5 is a schematic diagram of the biological analytical system of FIG. 4, illustrating the delivery of mixing bubbles of air to the RBC bath via a mixing pump to assist in the production of the final RBC solution. [Figure 5E] FIG. 5 is a schematic diagram of the biological analytical system of FIG. 4, illustrating the delivery of diluent from the sweep flow tank to the complete blood count (CBC) chamber of the system. [Figure 5F] FIG. 5 is a schematic diagram of the biological analytical system of FIG. 4, illustrating the filling of the diluent pump with diluent. [Figure 5G] FIG. 5 is a schematic diagram of the biological analytical system of FIG. 4, showing the use of diluent to flush the RBC bath via the wash pump, the diluent pump, and the waste pump to purify the waste fluid contained in the RBC bath from the RBC bath. [Figure 5H] FIG. 5 is a schematic diagram of the biological analysis system of FIG. 4, showing flushing the WBC bath via a wash pump and a waste pump, and purging the WBC bath of waste fluid contained therein using a diluent. [Figure 5I] FIG. 5 is a schematic diagram of the biological analytical system of FIG. 4 illustrating further filling of the diluent pump with diluent and further illustrating the formation of an air gap through the diluent pump. [Figure 5J] FIG. 5 is a schematic diagram of the biological analytical system of FIG. 4, illustrating the delivery of diluent to the WBC bath via a diluent pump. [Figure 6] FIG. 5 illustrates a method of analyzing a biological sample using the biological analysis system of FIG. 4. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0009] The drawings are not intended to be limiting in any way, and it is contemplated that various embodiments of the invention may be embodied in a variety of ways, including those not necessarily shown in the drawings. The accompanying drawings, which are incorporated in and form a part of this specification, illustrate several aspects of the invention and, together with the description, serve to explain the principles of the invention. It is understood, however, that the invention is not limited to the precise configurations shown.

[0010] The following description of a specific example of the present invention should not be used to limit the scope of the present invention. Other examples, features, aspects, embodiments, and advantages of the present invention will become apparent to those skilled in the art from the following description. The description is, by way of example, one of the best modes contemplated for carrying out the present invention. As will be understood, the present invention is capable of other distinct and obvious aspects, none of which deviates from the present invention. Therefore, the drawings and description should be regarded as illustrative in nature, rather than restrictive.

[0011] It will be appreciated that one or more of the teachings, formulas, versions, examples, etc. described herein may be combined with one or more of the other teachings, formulas, versions, examples, etc. described herein. Thus, the teachings, formulas, versions, examples, etc. described below should not be viewed in isolation from each other. Various suitable ways in which the teachings herein can be combined will be readily apparent to those skilled in the art in light of the teachings herein. Such modifications and variations are intended to be included within the scope of the claims.

[0012] Furthermore, the terms "about," "approximately," and the like, as used herein in connection with any numerical value or range of values, are intended to encompass, in addition to the exact value referenced, a suitable tolerance that allows the recited feature or combination of features to function for the intended purpose described herein.

[0013] The present disclosure relates to devices, systems, configurations and methods for analyzing biological samples, a first exemplary biological analysis system (10) and method (100) are described in detail with reference to Figures 1-2I and 3, respectively, and a second exemplary biological analysis system (210) and method (300) are described in detail with reference to Figures 4-5J and 6, respectively.

[0014] A. First Exemplary Biological Analytical System Embodiment

[0015] FIG. 1 illustrates an exemplary biological analysis system (10) that includes, among other components, a pair of fluid analysis chambers including a first fluid analysis chamber in the form of a first bath (12) and a second fluid analysis chamber in the form of a second bath (14). As illustrated in FIG. 1, the first bath (12) is a white blood cell (WBC) or hemoglobin (HGB) bath, and the second bath (14) is a red blood cell (RBC) bath. In the illustrated example, each of the WBC and RBC baths (12, 14) is open to allow a sample probe (16) ( FIG. 2C ) of the system (10) to selectively access one or both baths (12, 14), such as to aspirate fluid from the baths and / or dispense fluid to the baths. In other embodiments, at least one of the WBC or RBC baths (12, 14) may be closed (e.g., such that the sample probe (16) does not have access to at least one of the baths). In any event, the WBC and RBC baths (12, 14) of this embodiment are contained within a shielded complete blood count (CBC) box (18).

[0016] The biological analysis system (10) is configured to analyze a biological sample. In some embodiments, the biological analysis system (10) is configured to analyze a blood sample (B) (FIG. 2C), and therefore the biological analysis system (10) may be referred to as a blood analysis system. In this regard, the WBC bath (12) of this embodiment includes a hemoglobin transducer (20) configured to measure the amount of hemoglobin present in a fluid medium contained in the WBC bath (12). For example, the hemoglobin transducer (20) may include a light source (e.g., a filtered light source) and an optical sensor configured to receive an optical signal emitted from the light source through the fluid medium contained in the WBC bath (12). In some embodiments, the WBC bath and the RBC bath (12, 14) may each be fluidly coupled to a CBC drip chamber (not shown) housed in a shielded CBC box (18) via corresponding input and output conduits with respective valves for selectively transporting fluidic media from one of the WBC baths or RBC baths (12, 14) to the CBC drip chamber and / or returning such fluidic media from the CBC drip chamber to the WBC bath or RBC bath (12, 14). Such a CBC drip chamber may be configured to measure a complete blood count of the fluidic media received from each bath (12, 14). In other embodiments, only one of the WBC baths or RBC baths (12, 14) (e.g., only the RBC bath (14)) may be fluidly coupled to such a CBC drip chamber. Although analysis of blood (e.g., impedance-based counting, optical techniques, and / or imaging) is shown and described herein, the biological analysis system (10) may analyze (and optionally image) a variety of fluids, including other bodily fluids, such as, but not limited to, synovial fluid, urine, bone marrow, etc.

[0017] As shown in FIG. 1, the biological analysis system (10) also includes a plurality of fluid reservoirs, including a first fluid reservoir in the form of a diluent reservoir (30) containing a diluent (D) and a second fluid reservoir in the form of a lysis fluid reservoir (32) containing a lysis fluid (L). The biological analytical system (10) further includes a plurality of fluid pumps, including a first fluid pump in the form of a wash pump (40) configured to transport diluent (D) from the diluent reservoir (30) to various components of the biological system (10) as well as various other functions described below, a second fluid pump in the form of a lysate pump (42) configured to transport lysate (L) from the lysate reservoir (32) to the WBC bath (12), and a third pump in the form of a waste pump (44) configured to transport one or more fluid media from the WBC and RBC baths (12, 14) (and / or from the CBC drip chamber) to a CBC waste container (46). In the example shown, the wash pump (40) is configured as a piston or syringe pump, and the lysate pump (42) and the waste pump (44) are each configured as metering pumps. However, it will be appreciated that pumps (40, 42, 44) may each be configured in any manner suitable to perform their respective functions described herein.

[0018] To that end, the biological analytical system (10) further includes a plurality of fluid conduits and associated valves for selectively placing the various components of the biological analytical system (10) in fluid communication / non-communication with one another. More particularly, the biological analytical system (10) includes a first fluid conduit in the form of a diluent supply line (50) extending from the diluent reservoir (30) to the wash pump (40), and a first valve in the form of a fill-and-wash pump valve (52) located along the diluent supply line (50) between the diluent reservoir (30) and the wash pump (40), for selectively placing the diluent reservoir (30) in fluid communication / non-communication with one another. The biological analytical system (10) also includes a second fluid conduit in the form of a common RBC / WBC input line (54) extending from a first T-junction (56) located along the diluent supply line (50) between the wash pump (40) and the fill-and-wash pump valve (52) to a second T-junction (58). The biological analytical system (10) further includes a third fluid conduit in the form of an RBC input line (60) extending from the second T-junction (58) to the RBC bath (14), and a second valve in the form of a wash RBC valve (62) located along the RBC input line (60) between the RBC bath (14) and the wash pump (40) for selectively placing the RBC bath (14) in fluid communication / de-communication with the wash pump (40). The biological analysis system (10) also includes a fourth fluid conduit in the form of a first WBC input line (64) extending from the second T-junction (58) to the WBC bath (12) for selectively placing the WBC bath (12) in fluid communication / non-communication with the wash pump (40), and a third valve in the form of a wash WBC valve (66) located along the first WBC input line (64) between the WBC bath (12) and the wash pump (40).

[0019] The biological analytical system (10) further includes a fifth fluid conduit in the form of a lysate supply line (68) extending from the lysate reservoir (32) to the lysate pump (42) for selectively placing the lysate reservoir (32) in fluid communication with the lysate pump (42), and a fourth valve in the form of a lysate valve (70) located along the lysate supply line (68) between the lysate reservoir (32) and the lysate pump (42). The biological analytical system (10) also includes a sixth fluid conduit in the form of a second WBC input line (72) extending from the lysate pump (42) to the WBC bath (12) for selectively placing the lysate pump (42) in fluid communication with the WBC bath (12).

[0020] The biological analytical system (10) further includes seventh and eighth fluid conduits in the form of RBC and WBC drain lines (74, 76) extending from the RBC bath (14) and WBC bath (16), respectively, to a third T-junction (78), and a ninth fluid conduit in the form of a common RBC / WBC drain line (80) extending from the third T-junction (78) to the waste pump (44). The biological analytical system (10) also includes fifth and sixth valves in the form of drain RBC and drain WBC valves (82, 84) located along the RBC and WBC drain lines (74, 76), respectively, for selectively placing the corresponding baths (12, 14) in / out of fluid communication with the waste pump (44).

[0021] 2A-2I, the biological analytical system (10) is configured to direct various fluid media to predetermined components of the biological analytical system (10) to facilitate analysis of a blood sample (B) or other biological sample. It will be recognized that the open and / or closed states of the various valves of the biological analytical system (10) depicted are for illustrative purposes only, and that the actual states of the valves may differ from those depicted.

[0022] As shown in FIG. 2A , the wash pump (40) is configured to draw diluent (D) from the diluent reservoir (30) into the diluent supply line (50), such as when the fill-and-wash pump valve (52) is in an open state, and to transport diluent (D) to the WBC and RBC baths (12, 14) via the common RBC / WBC input line (54) and the respective RBC and WBC input lines (60, 64), such as when the fill-and-wash pump valve (52) is in a closed state and the wash RBC and wash WBC valves (62, 66) are each in an open state. In the event that one or both of the WBC and RBC baths (12, 14) may each initially contain waste fluid (W), which may be left over from a previous procedure, the waste pump (44) may be configured to draw waste fluid (W) and / or diluent (D) from the WBC and RBC baths (12, 14) into the corresponding RBC and WBC drain lines (74, 76) and further into the common RBC / WBC drain line (80) for drainage into the waste container (46), such as when the drain RBC and drain WBC valves (82, 84), respectively, are in an open state. In this manner, the wash pump (40) and the waste pump (44) may be configured to cooperate with one another to flush the waste fluid (W) from the WBC and RBC baths (12, 14) and clean and empty the WBC and RBC baths (12, 14).

[0023] 2B, the wash pump (40) is also configured to deliver diluent (D) to the empty WBC bath (12) via the common RBC / WBC input line (54) and the first WBC input line (64), such as when the fill-wash pump valve (52) is in a closed state, the wash WBC valve (66) is in an open state, and the wash RBC valve (62) is in a closed state. For example, the wash pump (40) may be configured to deliver a predetermined amount of diluent (D) to the WBC bath (12), such as about 500 μL to about 1000 μL, or about 800 μL of diluent (D).

[0024] As shown in FIG. 2C, the probe (16) is configured to dispense the blood sample (B) into the WBC bath (12) for mixing with the diluent (D) contained in the WBC bath (12), such as when the wash WBC valve (66) is in a closed state. For example, the probe (16) may be configured to deliver a predetermined amount of the blood sample (B) to the WBC bath (12), such as about 5 μL to about 25 μL, about 10 μL to about 15 μL, or about 12.4 μL of the blood sample (B). In this regard, the probe (16) may be configured to aspirate the blood sample (B) from a specimen (not shown) at a home or rest position and travel along a path from the home position to the WBC bath (12) to dispense the blood sample (B) into the WBC bath (12). In some embodiments, the path may not include an RBC bath (14). For example, the probe (16) may be configured to return to a home position from the WBC bath (12) without reaching the RBC bath (14).

[0025] 2D, the wash pump (40) is further configured to deliver additional diluent (D) to the WBC bath (12) via the common RBC / WBC input line (54) and the first WBC input line (64) for mixing with the blood sample (B) and diluent (D) already contained in the WBC bath (12), such as when the fill-wash pump valve (52) is in a closed state, the wash WBC valve (66) is in an open state, and the wash RBC valve (62) is in a closed state. For example, the wash pump (40) may be configured to deliver a predetermined amount of additional diluent (D) to the WBC bath (12), such as about 500 μL to about 1000 μL, or about 800 μL of additional diluent (D). In some embodiments, the wash pump (40) may be configured to deliver additional diluent (D) concomitantly to the blood sample (B) and diluent (D) already contained in the WBC bath (12) to facilitate mixing of the contents of the WBC bath (12) (e.g., both deliveries of blood sample (B) and diluent (D)) into a homogenous state defining a pre-diluent solution (P).

[0026] 2E, the wash pump (40) is also configured to draw at least diluent (D) contained within the common RBC / WBC input line (54) and the first WBC input line (64) in an upstream direction, such as when the fill-wash pump valve (52) is in a closed state, the wash WBC valve (66) is in an open state, and the wash RBC valve (62) is in a closed state, thereby drawing an extract portion (E) of pre-diluent solution (P) from the WBC bath (12) into the first WBC input line (64) and the common RBC / WBC input line (54). For example, the wash pump (40) may be configured to draw a predetermined amount of pre-diluent solution (P) from the WBC bath (12), such as about 200 μL to about 1000 μL, about 400 μL to about 800 μL, or about 600 μL of pre-diluent solution (P), to define the extract portion (E). In some embodiments, the upstream end of the extraction portion (E) may contact and be diluted by the diluent (D) in the common RBC / WBC input line (54) and / or the first WBC input line (64), thus defining a breakage region (C) of the extraction portion (E). For example, the breakage region (C) may not be homogenous (e.g., composed almost entirely of diluent instead of a homogenous mixture of blood sample and diluent) and / or may not represent the pre-diluent solution (P) contained within the WBC bath (12). Thus, the wash pump (40) may be configured to withdraw the broken region (C) substantially upstream of the second T-junction (58) such that the broken region (C) is entirely into the common RBC / WBC input line (54) and at least a portion of the unbroken region of the extraction portion (E) (e.g., representing the pre-diluent solution (P) that is homogenous and contained within the WBC bath (12)) is withdrawn upstream of the second T-junction (58) into the common RBC / WBC input line (54) to define the transport portion (T) of the pre-diluent solution (P). For example, the wash pump (40) may be configured to withdraw a predetermined amount of the unbroken region of the extraction portion (E), such as about 10 μL to about 100 μL, or about 60 μL of the unbroken region of the extraction portion (E), into the common RBC / WBC input line (54) to define the transport portion (T) of the pre-diluent solution (P).

[0027] 2F, the wash pump (40) is further configured to push at least the diluent (D) contained in the common RBC / WBC input line (54) in a downstream direction, such as when the fill-wash pump valve (52) is in a closed state, the wash WBC valve (66) is in a closed state, and the wash RBC valve (62) is in an open state, thereby conveying a transport portion (T) of the pre-diluent solution (P) from the common RBC / WBC input line (54) to the RBC input line (60) and conveying at least a portion of the diluent (D) contained in the RBC input line (60) to the empty RBC bath (14). For example, the wash pump (40) may be configured to deliver a predetermined amount of diluent (D) to the RBC bath (14), such as about 10 μL to about 100 μL, or about 60 μL of diluent (D), substantially equal to the predetermined amount of pre-diluent solution (P) that defines the transport portion (T) delivered to the RBC input line (60).

[0028] As shown in FIG. 2G, the lysing fluid pump (42) is configured to draw lysing fluid (L) from the lysing fluid reservoir (32) into the lysing fluid supply line (68), such as when the lysing fluid valve (70) is in an open state, and deliver the lysing fluid (L) to the WBC bath (12) via the second WBC input line (72) for mixing with the portion of the prediluent solution (P) remaining in the WBC bath (12). For example, the lysing fluid pump (42) may be configured to deliver a predetermined amount of lysing fluid (L) to the WBC bath (12), such as about 250 μL to about 750 μL, about 400 μL to about 500 μL, or about 453 μL of lysing fluid (L). In one example, the lysing fluid (L) is utilized to disrupt red blood cells in the WBC bath (12) and leave white blood cells (WBCs) for later analysis (e.g., via a CBC drip chamber).

[0029] As shown in FIG 2H, the wash pump (40) is further configured to deliver additional diluent (D) to the WBC bath (12) via the common RBC / WBC input line (54) and the first WBC input line (64) for mixing with the portion of lysing solution (L) and prediluent solution (P) already contained in the WBC bath (12), such as when the fill-wash pump valve (52) is in a closed state, the wash WBC valve (66) is in an open state, and the wash RBC valve (62) is in a closed state. For example, the wash pump (40) may be configured to deliver a predetermined amount of additional diluent (D) to the WBC bath (12), such as about 1,000 μL to about 2,000 μL, about 1,500 μL to about 1,800 μL, or about 1,600 μL of additional diluent (D). In some embodiments, the wash pump (40) may be configured to deliver additional diluent (D) concomitantly to the lysing fluid (L) and prediluent solution (P) already contained in the WBC bath (12) to facilitate mixing of the contents of the WBC bath (12) (e.g., the lysing fluid (L), the prediluent solution (P), and the additional diluent (D)) into a homogenous state that defines the final WBC solution (FW). In some embodiments, the final WBC solution (FW) may have a volume of about 3,005.5 μL, which may include about 11.9 μL of blood sample (B), about 2,540.6 μL of diluent (D), and about 453 μL of lysing fluid. In any event, the hemoglobin transducer (20) of the WBC bath (12) is configured to measure the amount of hemoglobin present in the final WBC solution (FW). For example, the hemoglobin transducer (20) may be configured to measure the amount of hemoglobin present in the final WBC solution (FW) upon completion of a hemoglobin incubation period, which may begin at the beginning of delivery of lysing solution (L) to the WBC bath (12). In some embodiments, the biological analysis system (10) may be configured to initiate white blood cell count flow priming and / or data acquisition from the final WBC solution (FW) (e.g., using a CBC drip chamber) upon completion of a WBC incubation period, such as about 9.5 seconds, which may also begin at the beginning of delivery of lysing solution (L) to the WBC bath (12).In one example, a lysing solution (L) is used in the WBC bath (12) to remove red blood cells (RBCs), thereby exposing the hemoglobin contained in the RBCs for analysis via a hemoglobin transducer (20).

[0030] 2I, the wash pump (40) is also configured to deliver the additional diluent, as well as a transfer portion (T) of the pre-diluent solution (P) contained in the RBC input line (60), to the RBC bath (14) via the common RBC / WBC input line (54) and the RBC input line (60), such as when the fill-wash pump valve (52) is in a closed state, the wash WBC valve (66) is in a closed state, and the wash RBC valve (62) is in an open state. For example, the wash pump (40) may be configured to deliver a predetermined amount of additional diluent (D) to the RBC bath (14), such as about 2,000 μL to about 3,000 μL, or about 2,800 μL of additional diluent (D). In some embodiments, the wash pump (40) may be configured to deliver additional diluent (D) and / or the transfer portion (T) of the pre-diluent solution (P) concomitantly to the diluent (D) already contained in the RBC bath (14) to facilitate mixing of the contents of the RBC bath (14) (e.g., both the transfer portion (T) of the pre-diluent solution (P) and the delivery of diluent (D)) into a homogenous state that defines the final RBC solution (FR). In some embodiments, the final RBC solution (FR) may have a volume of about 2,860 μL, which may include about 0.5 μL of blood sample (B) and about 2,859.5 μL of diluent (D). Additionally or alternatively, the final RBC solution (FR) may have a diluent to blood ratio of about 6,251. In some embodiments, the biological analytical system (10) may be configured to initiate red blood cell / platelet count flow priming and / or data acquisition from the Final RBC Solution (FR) (e.g., using a CBC drip chamber) upon completion of mixing of the Final RBC Solution (FR) to a homogenous state.

[0031] Although not shown, the biological analysis system (10) may further include various other suitable components for facilitating analysis of the blood sample (B) or other biological sample. For example, the biological analysis system (10) may further include a sweep flow tank (not shown) fluidly coupled to the first T-junction (56) and / or the RBC bath (14) via corresponding conduits with respective valves for selectively transporting a fluid medium therebetween. Additionally or alternatively, the biological analysis system (10) may include a CBC gas pump (not shown) fluidly coupled to the CBC drip chamber via one or more corresponding conduits with respective valves for selectively transporting a fluid medium therebetween. In some embodiments, the biological analysis system (10) may include at least one heater (not shown) configured to heat the diluent (D) (e.g., in the common RBC / WBC input line (54)) and / or the lysing solution (L) (e.g., in the second WBC input line (72)).

[0032] It will be appreciated that by utilizing the wash pump (40) to aspirate an extracted portion of the pre-diluent solution (P) from the WBC bath (12) and / or by utilizing the wash pump (40) to deliver a transport portion (T) of the extracted portion (E) of the pre-diluent solution (P) to the RBC bath (14), the biological analytical system (10) may be less reliant on the probe (16) to facilitate analysis of the blood sample (B) in the RBC bath (14) as compared to systems that utilize a probe (16) to at least aspirate a portion of the pre-diluent solution (P) from the WBC bath (12) and / or dispense an aspirated portion of the pre-diluent solution (P) into the RBC bath (14). Thus, the probe (16) may be configured to perform various other tasks while the wash pump (40) aspirates the extraction portion (E) of the pre-diluent solution (P) from the WBC bath (12) and / or while the wash pump (40) delivers the transport portion (T) of the extraction portion (E) of the pre-diluent solution (P) to the RBC bath (14). For example, the probe (16) may be configured to facilitate analysis of various other parameters, such as flow imaging of RBCs, WBCs, and PLTs, while the wash pump (40) facilitates delivery of the transport portion (T) to the RBC bath (14).

[0033] It will also be appreciated that by removing the RBC bath (14) from the path of the probe (16), the biological analytical system (10) may reduce the complexity and / or footprint of the path of the probe (16) at least as compared to systems that include an RBC bath (14) along the path of the probe (16), thereby allowing the biological analytical system (10) to have a reduced overall size at least as compared to such systems.

[0034] Although the embodiments herein have described the WBC bath (12) and RBC bath (14) as part of a shielded CBC box (18) connected to a CBC drip chamber, other embodiments are contemplated that utilize the pumped fluid transport technology contemplated herein. For example, the WBC bath (12) and RBC bath (14) can be used in a blood analysis system that utilizes flow imaging, where, for example, the WBC bath (12) and RBC bath (14) are connected to a flow cell and the contents of each are transported through the flow cell for imaging with a high-speed, high-resolution camera. The principle of using a pump (40) to facilitate fluid transport between chambers or baths of an analysis system to reduce reliance on a probe (16) can be widely used to reduce system load and increase throughput in a variety of situations.

[0035] It should be noted that other embodiments (e.g., systems and associated methods) utilize the wash pump / syringe pump fluid transport system discussed herein. For example, a probe (16) can first deliver the diluent and blood sample to the RBC bath (14), and the sample mixture / pre-diluent solution is then transported to the WBC bath (12) via pump (40). Other embodiments can utilize more than two baths (12, 14), in which case pump (40) is used to transport the blood sample solution (e.g., blood and diluent mixture) between one or more of the more than two baths. For example, rather than integrating the HGB transducer (20) with the WBC bath (12) for hemoglobin measurement purposes, a third bath can be utilized for this purpose.

[0036] B. First Exemplary Method of Analyzing a Biological Sample

[0037] 3 illustrates an exemplary method (100) of analyzing a biological sample, such as a blood sample (B). Although the method (100) is described as being performed using the biological analytical system (10) described above, it will be appreciated that the method (100) may be performed using any suitable biological analytical system.

[0038] Method (100) begins with step (101), in which first and second fluid analysis chambers, such as WBC and RBC baths (12, 14), are flushed, thereby cleaning, and / or emptied of any waste fluid (W) contained therein. In some embodiments, step (101) may be performed by drawing diluent (D) from a diluent reservoir (30) into a diluent supply line (50) and conveying the diluent (D) to the WBC and RBC baths (12, 14) via a wash pump (40) and / or by draining the waste fluid (W) and / or diluent (D) from the WBC and RBC baths (12, 14) via a waste pump (44), as described above in connection with FIG. 2A. It will be appreciated that at the completion of step (101), each of the common RBC / WBC input line (54), the RBC input line (60), and the first WBC input line (64) are substantially filled with clean diluent (D), and each of the WBC and RBC baths (12, 14) are substantially empty.

[0039] Method (100) proceeds from step (101) to step (102), in which diluent (D) is pumped or otherwise transported to a first fluid analysis chamber, such as HGB / WBC bath (12). In some embodiments, step (102) may be performed by delivering a predetermined volume of diluent (D), such as about 2 mL of diluent (D), to the WBC bath (12) via wash pump (40), as described above in connection with FIG. 2B.

[0040] From step (102), method (100) proceeds to step (103), where blood sample (B) is dispensed into a first fluid analysis chamber, such as an HGB / WBC bath (12). In some embodiments, step (103) may be performed by delivering a predetermined amount of blood sample (B), such as about 10 μL of blood sample (B), via probe (16) to WBC bath (12) for mixing with diluent (D) contained in WBC bath (12), as described above in connection with FIG. 2C. It will be appreciated that prior to step (103), method (100) may include aspirating blood sample (B) from a specimen (not shown) via probe (16) in a home position, and then moving probe (16) along a path from the home position to WBC bath (12) to dispense blood sample (B) into WBC bath (12). The method (100) may also include returning the probe (16) to a home position from the WBC bath (12) without reaching the RBC bath (14).

[0041] From step (103), method (100) proceeds to step (104), in which additional diluent (D) is pumped or otherwise conveyed to a first fluid analysis chamber, such as WBC bath (12). In some embodiments, step (104) may be performed by delivering a predetermined amount of additional diluent (D), such as about 800 μL of additional diluent (D), via wash pump (40) to WBC bath (12) concomitantly to the blood sample (B) and diluent (D) already contained in WBC bath (12) to facilitate mixing of the contents of WBC bath (12) into a homogenous state defining a pre-diluent solution (P), as described above in connection with FIG. 2D.

[0042] Method (100) proceeds from step (104) to step (105), in which an extracted portion (E) of pre-dilution solution (P) is pumped or otherwise aspirated from a first fluid analysis chamber, such as an HGB / WBC bath (12). In some embodiments, step (105) may be performed by withdrawing a predetermined amount of pre-diluent solution (P), such as about 200 μL of pre-diluent solution (P), from the HGB / WBC bath (12) via the wash pump (40) to define an extraction portion (E), and withdrawing a broken region (C) of the extraction portion (E) substantially upstream of the second T-junction (58) such that a predetermined amount of the unbroken region of the extraction portion (E), such as about 60 μL of the unbroken region of the extraction portion (E), is withdrawn upstream of the second T-junction (58) into the common RBC / WBC input line (54) to define a transport portion (T) of the pre-diluent solution (P), as described above in connection with FIG. 2E. Thus, step (105) can be performed without utilizing probe (16) to aspirate the transport portion (T) of pre-dilution solution (P) from the WBC bath (16), thereby freeing probe (16) to perform other tasks simultaneously with the performance of step (105).

[0043] From step (105), method (100) proceeds to step (106), in which a transport portion (T) of pre-diluent solution (P) is pumped or otherwise conveyed to a delivery conduit of a second fluid analysis chamber, such as an RBC input line (60). In some embodiments, step (106) may be performed by delivering the transport portion (T) to the RBC input line (60) via a wash pump (40), as described above in connection with FIG. 2F. It will be appreciated that a predetermined amount of diluent (D) contained in the RBC input line (60), such as about 60 μL of diluent (D), may be delivered to the empty RBC bath (14) via the wash pump (40) during performance of step (106).

[0044] From step (106), method (100) proceeds to step (107), in which lysis solution (L) is pumped or otherwise transported to a first fluid analysis chamber, such as WBC bath (12). In some embodiments, step (107) may be performed by delivering a predetermined amount of lysis solution (L), such as about 288 μL of lysis solution (L), to the WBC bath (12) for mixing with the portion of pre-diluent solution (P) remaining in the WBC bath (12) via lysis solution pump (42), as described above in connection with FIG. 2G.

[0045] From step (107), method (100) proceeds to step (108), in which additional diluent (D) is pumped or otherwise conveyed to a first fluid analysis chamber, such as WBC bath (12). In some embodiments, step (108) may be performed by delivering a predetermined amount of additional diluent (D), such as about 1,600 μL of additional diluent (D), via wash pump (40) to WBC bath (12) concomitantly to the lysis solution (L) and pre-diluent solution (P) already contained in WBC bath (12) to facilitate mixing of the contents of WBC bath (12) to a homogenous state that defines the final WBC solution (FW), as described above in connection with FIG. 2H.

[0046] From step (108), method (100) proceeds to step (109), in which the additional diluent (D) and the transport portion (T) of the pre-diluent solution (P) are pumped or otherwise conveyed to a second fluid analysis chamber, such as an RBC bath (14). In some embodiments, step (109) may be performed by delivering the transport portion (T) of the pre-diluent solution (P) and a predetermined amount of additional diluent (D), such as about 2,800 μL of additional diluent (D), to the RBC bath (14) via wash pump (40), concomitantly to the diluent (D) already contained in the RBC bath (14), to facilitate mixing of the contents of the RBC bath (14) to a homogenous state that defines the final WBC solution (FW), as described above in connection with FIG. 2I. Thus, step (109) can be performed without utilizing probe (16) to deliver the transport portion (T) of pre-dilution solution (P) to the RBC bath (14), thereby freeing probe (16) to perform other tasks simultaneously with the performance of step (109).

[0047] From step (109), method (100) proceeds to step (110), where the Final RBC Solution (FR) in a second fluid analysis chamber, such as RBC bath (14), is analyzed. In some embodiments, step (110) may occur immediately following completion of step (109). In any event, step (110) may include initiating red blood cell / platelet count flow priming and / or data acquisition from the Final RBC Solution (FR).

[0048] In the illustrated example, the method (100) also proceeds from step (108) to step (111), where the final WBC solution (FW) in a first fluid analysis chamber, such as the WBC bath (12), is analyzed. In some embodiments, step (111) may occur upon completion of one or more incubation periods, which may begin upon initiation of delivery of lysing fluid (L) to the WBC bath (12) during performance of step (107). For example, step (111) may include measuring the amount of hemoglobin present in the final WBC solution (FW), such as with a hemoglobin transducer (20) of the WBC bath (12), upon completion of the hemoglobin incubation period. Additionally or alternatively, step (111) may include initiating white blood cell count flow priming and / or data acquisition from the final WBC solution (FW) upon completion of the WBC incubation period, such as approximately 12-14 seconds from initiation of delivery of lysing fluid (L) to the WBC bath (12). It will be appreciated that step (111) may be performed in parallel with one or more of steps (109, 110).

[0049] In some embodiments, after steps (110, 111) are completed, the method (100) may return to step (101), where the first and second fluid analysis chambers, such as the WBC bath and RBC bath (12, 14), are flushed, thereby cleaned, and / or emptied of any waste fluid (W) contained therein (e.g., the final WBC and RBC solutions (FW, FR) from the previous cycle), in preparation for a new cycle.

[0050] C. Second Exemplary Biological Analytical System Embodiment

[0051] FIG. 4 illustrates another exemplary biological analysis system (210) that includes, among other components, a pair of fluid analysis chambers including a first fluid analysis chamber in the form of a first bath (212) and a second fluid analysis chamber in the form of a second bath (214). As shown in FIG. 4, the first bath (212) is a white blood cell (WBC) or hemoglobin (HGB) bath, and the second bath (214) is a red blood cell (RBC) bath. In the illustrated example, the WBC bath (212) is open to allow a sample probe (216) of the system (210) to selectively access the WBC bath (212), such as to aspirate fluid from the bath and / or dispense fluid to the bath. Although not illustrated, the WBC and RBC baths (212, 214) of this embodiment may be housed in a shielded complete blood count (CBC) box similar to the CBC box (18).

[0052] The biological analysis system (210) is configured to analyze a biological sample. In some embodiments, the biological analysis system (210) is configured to analyze a blood sample (B) (FIG. 5A), and therefore the biological analysis system (210) may be referred to as a blood analysis system. Although not shown, the WBC bath (212) of this embodiment may include a hemoglobin transducer, similar to the hemoglobin transducer (20), configured to measure the amount of hemoglobin present in the fluid medium contained in the WBC bath (212). For example, the hemoglobin transducer may include a light source (e.g., a filtered light source) and an optical sensor configured to receive an optical signal emitted from the light source through the fluid medium contained in the WBC bath (212). In some embodiments, the WBC bath and the RBC bath (212, 214) may each be fluidly coupled to a CBC drip chamber (221) housed in a shielded CBC box via corresponding input and output conduits with respective valves for selectively transporting fluidic media from one of the WBC baths or RBC baths (212, 214) to the CBC drip chamber (221) and / or returning such fluidic media from the CBC drip chamber (221) to the WBC bath or RBC bath (212, 214). The CBC drip chamber (221) may be configured to measure a complete blood count of the fluidic media received from each bath (212, 214). In other embodiments, only one of the WBC baths or the RBC baths (212, 214) (e.g., only the RBC bath (214)) may be fluidly coupled to the CBC drip chamber (221). In the example shown, the CBC drip chamber (221) is also fluidly coupled to a pneumatic transducer (222). Although analysis of blood (e.g., impedance-based counting, optical techniques, and / or imaging) is shown and described herein, the biological analysis system (210) may analyze (and optionally image) a variety of fluids, including other bodily fluids, such as, but not limited to, synovial fluid, urine, bone marrow, etc.

[0053] As shown in FIG. 4, the biological analysis system (210) also includes a plurality of fluid reservoirs, including a first fluid reservoir in the form of a diluent reservoir (230) containing a diluent (D), a second fluid reservoir in the form of a lysis fluid reservoir (232) containing a lysis fluid (L), and a third fluid reservoir in the form of a cleaning agent reservoir (233) containing a cleaning agent (CL). The biological analytical system (210) further includes a plurality of fluid pumps, including a first fluid pump in the form of a wash pump (240) configured to transport diluent (D) from the diluent reservoir (230) and / or to transport clarifier (CL) from the clarifier reservoir (233) to various components of the biological system (210), including the sweep flow tank (241); a second fluid pump in the form of a lysing fluid pump (242) configured to transport lysing fluid (L) from the lysing fluid reservoir (232) to the WBC bath (212); and a third fluid pump in the form of a vacuum pump (244) configured to adjust the vacuum for a desired count via the pneumatic transducer (222). the third pump, a fourth pump in the form of a waste pump (245) configured to transport one or more fluid media from the WBC and RBC baths (212, 214) (and / or from the CBC drip chamber (221)) to a waste container (246); a fifth pump in the form of a mixing pump (247) configured to deliver mixed bubbles of air (A) from the atmosphere (248) to the WBC and RBC baths (212, 214); and a sixth pump in the form of a diluent pump (249) configured to transport diluent (D) from the sweep flow tank (241) to the WBC and RBC baths (212, 214) as well as perform various other functions described below. In the example shown, diluent pump (249) is configured as a piston or syringe pump, and wash pump (240), dissolver pump (242), vacuum pump (244), waste pump (245), and mix pump (247) are each configured as peristaltic pumps, however, it will be appreciated that pumps (240, 242, 244, 245, 247, 249) may each be configured in any manner suitable to perform their respective functions described herein.

[0054] To that end, the biological analytical system (210) further includes a plurality of fluid conduits and associated valves for selectively placing the various components of the biological analytical system (210) in fluid communication / non-communication with one another. More particularly, the biological analytical system (210) includes a fluid conduit in the form of a diluent supply line (250) extending from the diluent reservoir (230) through the wash pump (240) to the first multi-flow unit (251), and a valve in the form of a diluent / cleanant valve (252) located along the diluent supply line (250) between the diluent reservoir (230) and the wash pump (240), for selectively placing the diluent reservoir (230) and the first multi-flow unit (251) in fluid communication / non-communication with one another. The biological analysis system (210) also includes a fluid conduit in the form of a cleaning agent supply line (254) extending from the cleaning agent reservoir (233) to the diluent / cleanant valve (252) for cooperating with the diluent / cleanant valve (252) to selectively place the cleaning agent reservoir (233) and the first multi-flow unit (251) in fluid communication / non-communication with each other, and a valve in the form of a cleaning agent valve (255) located along the cleaning agent supply line (254) between the cleaning agent reservoir (233) and the diluent / cleanant valve (252).

[0055] The biological analysis system (210) also includes a fluid conduit in the form of an RBC input line (256) extending from the first multi-flow unit (251) to the RBC bath (214) for selectively placing the RBC bath (214) and the first multi-flow unit (251) in fluid communication / non-communication with each other, and a valve in the form of a washing RBC valve (257) located along the RBC input line (256) between the first multi-flow unit (251) and the RBC bath (214). The biological analysis system (210) also includes a fluid conduit in the form of a sweep flow input line (258) extending from the first multi-flow unit (251) to the sweep flow tank (241) for selectively placing the sweep flow tank (241) and the first multi-flow unit (251) in fluid communication / non-communication with each other, and a valve in the form of a cleaning sweep flow valve (259) located along the sweep flow input line (258) between the first multi-flow unit (251) and the sweep flow tank (241). The biological analysis system (210) further includes a fluid conduit in the form of a second multi-flow unit input line (260) extending from the first multi-flow unit (251) to the second multi-flow unit (261) for selectively placing the first and second multi-flow units (251, 261) in fluid communication / non-communication with one another, and a valve in the form of a multi-flow input valve (262) located along the second multi-flow unit input line (260) between the first and second multi-flow units (251, 261). The biological analysis system (210) also includes a fluid conduit in the form of a WBC input line (264) extending from the second multi-flow unit (261) to the WBC bath (212) for selectively placing the WBC bath (212) and the second multi-flow unit (261) in fluid communication / non-communication with each other, and a valve in the form of a washing WBC valve (265) located along the WBC input line (264) between the second multi-flow unit (261) and the WBC bath (212).

[0056] The biological analysis system (210) further includes a fluid conduit in the form of a diluent input line (266) extending from the sweep flow tank (241) to the diluent pump (249) for selectively placing the diluent pump (249) and the sweep flow tank (241) in fluid communication / non-communication with each other, and a valve in the form of a diluent input valve (267) located along the diluent input line (266) between the sweep flow tank (241) and the diluent pump (249). The biological analysis system (210) also includes a fluid conduit in the form of a common sweep flow input line (268) extending from the sweep flow tank (241) to the first Y-junction (269) for selectively placing the first Y-junction (269) and the sweep flow tank (241) in fluid communication / non-communication with each other, and a valve in the form of a sweep flow valve (270) located along the common sweep flow input line (268) between the sweep flow tank (241) and the first Y-junction (269). The biological analysis system (210) further includes fluid conduits in the form of sweep flow lines (271, 272) extending from the first Y-junction (269) to the CBC chamber (221) for selectively placing the CBC chamber (221) and the first Y-junction (269) in fluid communication / non-communication with each other, and valves in the form of sweep flow valves (273, 274) located along each of the sweep flow lines (271, 272) between the first Y-junction (269) and the CBC chamber (221).

[0057] The biological analysis system (210) also includes a fluid conduit in the form of a diluent input / output line (275) extending from the diluent pump (249) to the second multi-flow unit (261). The biological analysis system (210) further includes a fluid conduit in the form of a transport line including a plurality of transport line segments (276a, 276b, 276c, 276d) extending from the WBC bath (212) to the RBC bath (214) for selectively placing the plurality of transport line segments (276a, 276b, 276c, 276d) in fluid communication / non-communication with one another, and a plurality of valves in the form of first, second, and third transport line valves (277, 278, 279) located along the transport line (276) between the WBC bath and the RBC baths (212, 214). As shown, the first and second transfer line valves (277, 278) are each three-way valves. The second transfer line portion (276b) extending between the first and second transfer line valves (277, 278) may have a fixed, predetermined volume such that the second transfer line portion (276b) may be selectively and precisely filled with a fluid having the same predetermined volume. For example, the second transfer line portion (276b) may be constructed of a substantially rigid (e.g., non-expandable) tube. The biological analysis system (210) also includes fluid conduits in the form of diluent push lines (280) and diluent push / pull lines (281) extending from the second multi-flow unit (261) to the respective transport line valves (277, 278) such that the transport line valves (277, 278) can selectively place the respective diluent push line (280) or diluent push / pull line (281) in fluid communication / non-communication with the corresponding transport line portions (276a, 276b, 276c, 276d).

[0058] The biological analysis system (210) further includes a fluid conduit in the form of a lysing fluid supply line (282) that extends from the lysing fluid reservoir (232), through the lysing fluid pump (242), and to the WBC bath (212).

[0059] The biological analysis system (210) further includes a fluid conduit in the form of an RBC drain line (283) extending from the RBC bath (214) to the third multi-flow unit (284) for selectively placing the third multi-flow unit (284) and the RBC bath (214) in fluid communication / non-communication with each other, and a valve in the form of an RBC drain valve (285) located along the RBC drain line (283) between the RBC bath (214) and the third multi-flow unit (284). The biological analysis system (210) also includes a fluid conduit in the form of a mixed air line (286) extending from atmosphere (248), through the mixing pump (247), and along the RBC drain line (283) to a second Y-junction (287) located between the RBC bath (214) and the RBC drain valve (285) for selectively placing the RBC bath (214) and the mixing pump (247) in fluid communication / de-communication with each other, and a valve in the form of a mixing pump valve (288) located along the mixed air line (286) between the mixing pump (247) and the RBC bath (214). The biological analysis system (210) further includes a fluid conduit in the form of a WBC drain line (289) extending from the WBC bath (212) to the third multi-flow unit (284) for selectively placing the third multi-flow unit (284) and the WBC bath (212) in fluid communication / non-communication with each other, and a valve in the form of a WBC drain valve (290) located along the WBC drain line (289) between the WBC bath (212) and the third multi-flow unit (284). The biological analysis system (210) also includes a fluid conduit in the form of a mixing air line (291) extending from the mixing pump valve (288) along the WBC drain line (289) to a third Y-junction (292) located between the WBC bath (212) and the WBC drain valve (290), such that the mixing pump valve (288) can selectively place the WBC bath (212) and the mixing pump (247) in / out of fluid communication with each other.

[0060] The biological analysis system (210) also includes a fluid conduit in the form of a CBC drain line (293) extending from the CBC chamber (221) to the third multi-flow unit (284) for selectively placing the third multi-flow unit (284) and the CBC chamber (221) in fluid communication / non-communication with each other, and a valve in the form of a CBC drain valve (294) located along the CBC drain line (293) between the CBC chamber (221) and the third multi-flow unit (284). The biological analysis system (210) also includes a fluid conduit in the form of a waste line (295) extending from a waste container (246) through the vacuum pump (244) along the CBC drain line (293) to a fourth Y-junction (296) located between the CBC chamber (221) and the CBC drain valve (294). The biological analytical system (210) further includes a fluid conduit in the form of a waste line (297) that extends from the waste container (246), through the waste pump (245), and to the third multi-flow unit (284).

[0061] The biological analysis system (210) further includes a fluid conduit in the form of a vacuum line (298) extending from the CBC chamber (221) to the pneumatic pressure transducer (222) for selectively placing the pneumatic pressure transducer (222) and the CBC chamber (221) in fluid communication / non-communication with one another, and a valve in the form of a vacuum line valve (299) located along the vacuum line (298) between the CBC chamber (221) and the pneumatic pressure transducer (222). In some embodiments, the vacuum line valve (299) may be configured to selectively place the CBC chamber (221) in fluid communication / non-communication with the atmosphere (248), such as for degassing the CBC chamber (221).

[0062] 5A-5J, biological analytical system (210) is configured to direct various fluid media to predetermined components of biological analytical system (210) to facilitate analysis of a blood sample (B) or other biological sample. It will be recognized that the open and / or closed states of the various valves of biological analytical system (210) depicted are for illustrative purposes only, and that the actual states of the valves may differ from those depicted.

[0063] As shown in FIG. 5A , the wash pump (240) is configured to draw diluent (D) from the diluent reservoir (230) into the diluent supply line (250) (e.g., placing the diluent reservoir (230) and the first multi-flow unit (251) in fluid communication with each other), such as when the diluent / cleanant valve (252) is in a first open state, and to deliver diluent (D) through the first multi-flow unit (251) and the sweep flow input line (258) to the sweep flow tank (241) (e.g., placing the sweep flow tank (241) and the first multi-flow unit (251) in fluid communication with each other), such as when the wash sweep flow valve (259) is in an open state.

[0064] As also shown in FIG. 5A , the diluent pump (249) is configured to draw diluent (D) from the sweep flow tank (241) into the diluent input line (266) and further into the diluent pump (249) (e.g., placing the diluent pump (249) and the sweep flow tank (241) in fluid communication with each other) such that the diluent pump (249) can be filled with diluent (D), such as when the diluent input valve (267) is in an open state. In some embodiments, the diluent pump (249) may be configured to be filled with a first predetermined volume of diluent (D) via the diluent input line (266), such as the difference between the volume of the RBC bath (214) and the volume of pre-diluent solution (P) desired to subsequently extract from the WBC bath (212) to define the extraction portion (E), such as about 200 μL to about 250 μL of pre-diluent solution (P) (e.g., about 200 μL or about 250 μL of pre-diluent solution (P)), as described in further detail below.

[0065] As further shown in FIG. 5A, the probe (216) is configured to dispense the blood sample (B) into the WBC bath (212) for mixing with the diluent (D) initially contained in the WBC bath (212). For example, the probe (216) may be configured to deliver a predetermined amount of the blood sample (B) to the WBC bath (212), such as about 10 μL to about 16 μL of the blood sample (B) (e.g., about 10 μL, about 12 μL, about 14 μL, or about 16 μL of the blood sample (B)). In this regard, the probe (216) may be configured to aspirate the blood sample (B) from the specimen (not shown) at a home or rest position and travel along a path from the home position to the WBC bath (212) to dispense the blood sample (B) into the WBC bath (212). In some embodiments, the path may not include an RBC bath (214). For example, the probe (216) may be configured to return to a home position from the WBC bath (212) without reaching the RBC bath (214).

[0066] As also shown in FIG. 5A, the mixing pump (247) is configured to draw air (A) from the atmosphere (248) and deliver mixed bubbles of air (A) to the WBC bath (212) via the mixing air line (291) and the WBC drain line (289) (e.g., placing the WBC bath (212) and the mixing pump (247) in fluid communication with each other) to facilitate mixing of the contents of the WBC bath (212) (e.g., blood sample (B) and diluent (D)) into a homogenous state defining a pre-diluent solution (P), such as when the mixing pump valve (288) is in a first open state. As also shown in FIG. 5A, the waste pump (245) is configured to draw remaining waste fluid (W) from the CBC chamber (221) into the CBC drain line (293) and further into the waste line (295) to the waste container (246).

[0067] 5B, the diluent pump (249) is configured to draw diluent (D) from the second transport line portion (276b) through the diluent push / pull line (281) and the second multi-flow unit (261) into the diluent input / output line (275) and further into the diluent pump (249) so that the diluent pump (249) can be further filled with diluent (D). In some embodiments, the diluent pump (249) may be configured to be filled with a second predetermined volume of diluent (D) through the diluent input / output line (275). For example, as described in further detail below, the cumulative volume of diluent (D) that the diluent pump (249) is configured to fill via the diluent input line (266) and the diluent input / output line (275) may be selected to subsequently deliver a desired volume of diluent (D) to the RBC bath (214), such as from about 2.804 mL to about 3.364 mL of diluent (D) (e.g., about 2.804 mL or about 3.364 mL of diluent (D)).

[0068] As also shown in FIG. 5B , the diluent pump (249) is also configured to draw an air gap (G) from the first transport line portion (276a) through the second transport line portion (276a) into the diluent push / pull line (281), such as when the first and second transport line valves (277, 278) are each in a first open state, to draw the extracted portion (E) of the pre-diluent solution (P) from the WBC bath (212) into the first and second transport line portions (276a, 276b) and the diluent push / pull line (281) (e.g., placing the diluent push / pull line (281) in fluid communication with one another) with the first and second transport line portions (276a, 276b). For example, the diluent pump (249) may be configured to withdraw a predetermined amount of pre-diluent solution (P) from the WBC bath (212), such as about 200 μL to about 250 μL of pre-diluent solution (P) (e.g., about 200 μL or about 250 μL of pre-diluent solution (P)) to define the extraction portion (E). In some embodiments, the upstream end of the extraction portion (E) may be separated from the diluent (D) by an air gap (G) within the diluent push / pull line (281). In this manner, the air gap (G) may act as a protective barrier between the diluent (D) and the extraction portion (E) of the pre-diluent solution (P) to prevent the extraction portion (E) from being damaged by the diluent (D), thereby maintaining the integrity of the extraction portion (E). For example, the extraction portion (E) may remain homogenous and representative of the pre-diluent solution (P) contained in the WBC bath (212). Thus, the diluent pump (249) may be configured to draw a buffer region (BU) of the extraction portion (E) into the diluent push / pull line (281) such that the second transport line portion (276b) may be substantially filled with the transport portion (T) of the pre-diluent solution (P). For example, the diluent pump (249) may be configured to draw a predetermined amount of the extraction portion (E), such as about 100 μL to about 120 μL of the extraction portion (E) (e.g., about 100 μL or about 120 μL of the extraction portion (E)), into the second transport line portion (276b) to define the transport portion (T) of the pre-diluent solution (P).In this regard, the diluent pump (249), in cooperation with the first and second transfer line valves (277, 278), may be configured to completely fill the second transfer line portion (276b) with the transfer portion (T) such that the transfer portion (T) may have a predetermined volume substantially the same as the fixed predetermined volume of the second transfer line portion (276b).

[0069] As shown in FIG. 5C , the diluent pump (249) further provides a diluent input / output line (275), a diluent output line (276), and a diluent pump (249) for discharging the diluent from the diluent pump (249) when the first and second transfer line valves (277, 278) are each in a second open state (e.g., fluidly connecting the diluent push line (280) to the second and third transfer line portions (276b, 276c) with each other), and when the third transfer line valve (279) is in an open state (e.g., fluidly connecting the third and fourth transfer line portions (276c, 276d) with each other), etc. The diluent pump (249) is configured to convey the diluent (D) towards the second transport line portion (276b) via the second multi-flow unit (261) and the diluent push line (280), convey the transport portion (T) of the pre-diluent solution (P) from the second transport line portion (276b) via the third and fourth transport line portions (276c, 276d) to the RBC bath (214), and convey the diluent (D) from the third and fourth transport line portions (276c, 276d) to the RBC bath (214). For example, the diluent pump (249) may be configured to deliver a predetermined volume of diluent (D) to the RBC bath (214) that is substantially equal to a cumulative volume of diluent (D) that the diluent pump (249) has already been filled with. In some embodiments, the diluent pump (249) may be configured to concomitantly deliver the diluent (D) and / or the transferred portion (T) of the pre-diluent solution (P) to the RBC bath (214) to facilitate mixing of the contents of the RBC bath (214) (e.g., the transferred portion (T) of the pre-diluent solution (P) and the delivered diluent (D)) into a homogenous state that defines the final RBC solution (FR).

[0070] 5C, the lysing fluid pump (242) is configured to convey lysing fluid (L) from the lysing fluid reservoir (232) through the lysing fluid supply line (282) to the WBC bath (212) for mixing with the portion of the prediluent solution (P) remaining in the WBC bath (212). For example, the lysing fluid pump (242) may be configured to deliver a predetermined amount of lysing fluid (L) to the WBC bath (212), such as about 337.8 μL to about 575.9 μL of lysing fluid (L) (e.g., about 337.8 μL, about 346.6 μL, about 414.2 μL, about 423 μL, about 490.6 μL, about 499.5 μL, about 567 μL, or about 575.9 μL of lysing fluid (L)). In one example, a lysing solution (L) is utilized to destroy red blood cells in the WBC bath (212), leaving the white blood cells (WBCs) for later analysis (eg, via the CBC drip chamber (221)).

[0071] As further shown in FIG. 5C, the mixing pump (247) is configured to draw air (A) from the atmosphere (248) and deliver mixed bubbles of air (A) to the WBC bath (212) via the mixed air line (291) and the WBC drain line (289) to facilitate mixing of the contents of the WBC bath (212) (e.g., the prediluent solution (P) and the lysing solution (L)) into a homogenous state that defines the final WBC solution (FW), such as when the mixing pump valve (288) is in a first open state. A hemoglobin transducer in the WBC bath (212) is configured to measure the amount of hemoglobin present in the final WBC solution (FW). For example, the hemoglobin transducer may be configured to measure the amount of hemoglobin present in the final WBC solution (FW) upon completion of a hemoglobin incubation period, which may begin after the mixed bubbles of air (A) are delivered to the WBC bath (212). In some embodiments, the biological analysis system (210) may be configured to initiate white blood cell count flow priming and / or data acquisition from the final WBC solution (FW) (e.g., using the CBC drip chamber (221)) upon completion of a WBC incubation period, which may also begin after the mixed bubbles of air (A) are delivered to the WBC bath (212). In one example, a lysing solution (L) is used in the WBC bath (212) to remove red blood cells (RBCs), thereby exposing the hemoglobin contained within the RBCs for analysis via a hemoglobin transducer.

[0072] As shown in FIG. 5D , the mixing pump (247) is also configured to draw air (A) from the atmosphere (248) and deliver mixed bubbles of air (A) to the RBC bath (214) via the mixing air line (286) and the RBC drain line (283) (e.g., placing the RBC bath (214) and the mixing pump (247) in fluid communication with each other), such as when the mixing pump valve (288) is in a second open state, to facilitate mixing of the contents of the RBC bath (214) (e.g., the transferred portion (T) of the pre-diluent solution (P) and the delivered diluent (D)) to a homogenous state that defines the Final RBC Solution (FR). In some embodiments, the biological analysis system (210) may be configured to initiate red blood cell / platelet count flow priming and / or data acquisition from the Final RBC Solution (FR) (e.g., using the CBC drip chamber (221)) upon completion of mixing of the Final RBC Solution (FR) to a homogenous state.

[0073] As shown in FIG. 5E, the sweep flow tank (241) is configured to deliver diluent (D) via the common sweep flow input line (268) and the first Y-branch (269) to the sweep flow lines (271, 272) and further to the CBC chamber (221) (e.g., placing the sweep flow tank (241) and the CBC chamber (221) in fluid communication with each other), such as when the wash sweep flow valve (259) and the sweep flow valves (273, 274) are each in an open state. Also as shown in FIG. 5E, the waste pump (245) is configured to transport the remaining waste fluid (W) from the CBC chamber (221) via the CBC drain line (293) and the fourth Y-branch (296) to the waste line (295) and further to the waste container (246).

[0074] 5F, the diluent pump (249) is also configured to draw diluent (D) from the sweep flow tank (241) into the diluent input line (266) and further into the diluent pump (249) such that the diluent pump (249) may be filled with diluent (D), such as when the diluent input valve (267) is in an open state. In some embodiments, the diluent pump (249) may be configured to be filled with a third predetermined volume of diluent (D) via the diluent input line (266), such as a volume of diluent (D) desired for subsequently flushing the diluent input / output line (275), the diluent push line (280), and the second, third, and fourth transport line portions (276b, 276c, 276d), as described in further detail below.

[0075] As shown in FIG. 5G, the wash pump (240) is also configured to draw diluent (D) from the diluent reservoir (230) into the diluent supply line (250), such as when the diluent / cleanant valve (252) is in a first open state, and to transport diluent (D) through the first multi-flow unit (251) and the RBC input line (256) to the RBC bath (214) (e.g., placing the RBC bath (214) and the first multi-flow unit (251) in fluid communication with each other), such as when the wash RBC valve (257) is in an open state. 5G, the diluent pump (249) is also configured to deliver diluent (D) to the RBC bath (214) via the diluent input / output line (275), the second multi-flow unit (261), the diluent push line (280), and the second, third, and fourth transport line portions (276b, 276c, 276d), such as when the first and second transport line valves (277, 278) are each in a second open state and the third transport line valve (279) is in an open state. In some embodiments, the diluent pump (249) may be configured to deliver diluent (D) incidental to the diluent (D) delivered by the wash pump (240). Additionally or alternatively, the diluent pump (249) may be configured to draw diluent (D) from the sweep flow tank (241) into the diluent input line (266) and further into the diluent pump (249) such that the diluent pump (249) may then be filled with diluent (D), such as when the diluent input valve (267) is in an open state. In some embodiments, the diluent pump (249) may be configured to be filled with a fourth predetermined volume of diluent (D) via the diluent input line (266), such as the difference between the volume of the WBC bath (212) and the volume of the air gap (G).

[0076] As further shown in FIG. 5G, the waste pump (245) is configured to transport remaining waste fluid (W) from the RBC bath (214), through the RBC drain line (283) and the third multi-flow unit (284), into the waste line (297), and further to the waste container (246), such as when the RBC drain valve (285) is in an open state (e.g., placing the third multi-flow unit (284) and the RBC bath (214) in fluid communication with each other).

[0077] As shown in FIG. 5H , the wash pump (240) further draws diluent (D) from the diluent reservoir (230) to the diluent supply line (250), such as when the diluent / cleanant valve (252) is in a first open state, to the second multi-flow unit (261) via the first multi-flow unit (251) and the second multi-flow unit input line (260), such as when the multi-flow input valve (262) is in an open state (e.g., placing the first and second multi-flow units (251, 261) in fluid communication with each other), and to the second multi-flow unit (261) via the first multi-flow unit (251) and the second multi-flow unit input line (260), such as when the wash WBC valve (265) is in an open state (e.g., placing the first and second multi-flow units (251, 261) in fluid communication with each other). The diluent (D) is configured to convey diluent (D) via the WBC input line (264), such as when the first and second transport line valves (278) are each in a closed state, and / or via the diluent push / pull line (281) and the first and second transport line portions (276a, 276b) to the WBC bath (212), such as when the first and second transport line valves (277, 278) are each in a first open state and the wash WBC valve (265) is in a closed state (e.g., placing the WBC bath (212) and the second multi-flow unit (261) in fluid communication with each other). In some embodiments, the wash pump (240) may be configured to deliver diluent (D) via the first transport line portion (276a) incidental to the diluent (D) delivered via the WBC input line (264).

[0078] As further shown in FIG. 5H, the waste pump (245) is configured to transport remaining waste fluid (W) from the WBC bath (212) through the WBC drain line (289) and the third multi-flow unit (284), into the waste line (297), and further to the waste container (246), such as when the WBC drain valve (290) is in an open state (e.g., placing the third multi-flow unit (284) and the WBC bath (212) in fluid communication with each other).

[0079] 5I, the diluent pump (249) is further configured to draw diluent (D) from the WBC bath (212), through the first and second transport line portions (276a, 276b), the diluent push / pull line (281), and the second multi-flow unit (261), into the diluent input / output line (275), and further into the diluent pump (249), such that the diluent pump (249) may be filled with diluent (D), such as when the first and second transport line valves (277a, 278) are each in a first open state. In some embodiments, the diluent pump (249) may be configured to be filled with a fifth predetermined volume of diluent (D) through the diluent input / output line (275). For example, the cumulative volume of diluent (D) that the diluent pump (249) is configured to fill via the diluent input line (266) and the diluent input / output line (275) may be substantially equal to the volume of diluent (D) desired to subsequently deliver to the WBC bath (212), such as from about 2.143 mL to about 3.429 mL of diluent (D) (e.g., about 2.143 mL, about 2.571 mL, about 3 mL, or about 3.429 mL of diluent (D)), as described in further detail below.

[0080] As also shown in FIG. 5I, the diluent pump (249) is also configured to draw air from the WBC bath (212) into the first transport line portion (276a), such as when the first and second transport line valves (277a, 278) are each in a first open state, thereby creating an air gap (G) in the first transport line portion (276a) to act as a protective barrier between the diluent (D) and the extraction portion (E) of the pre-diluent solution (P) in a subsequent cycle.

[0081] As further shown in FIG. 5I, the CBC chamber (221) is configured to be evacuated (e.g., to atmosphere (248)), such as when the vacuum line valve (299) is in a first open position (e.g., placing the CBC chamber (221) and the atmosphere (248) in fluid communication with each other).

[0082] As shown in FIG. 5J, the diluent pump (249) is further configured to convey diluent (D) via the diluent input / output line (275), the second multi-flow unit (261) and the WBC input line (264) to the WBC bath (212), such as when the wash WBC valve (265) is in an open state, for mixing with the blood sample (B) in a subsequent cycle to form a pre-diluent solution (P). As also shown in FIG. 5I, the waste pump (245) is configured to convey remaining waste fluid (W) from the CBC chamber (221), via the CBC drain line (293) and the third multi-flow unit (284), into the waste line (297) and further to the waste container (246), such as when the CBC drain valve (294) is in an open state (e.g., placing the third multi-flow unit (284) and the CBC chamber (221) in fluid communication with each other). As further shown in FIG. 5J, the CBC chamber (221) is configured to be pressurized by the pneumatic pressure transducer (222) (e.g., placing the pneumatic pressure transducer (222) and the CBC chamber (221) in fluid communication with each other), such as when the vacuum line valve (299) is in a second open state.

[0083] The biological analytical system (210) may be configured to complete a subsequent cycle by returning to the state shown in FIG. 5A.

[0084] It will be appreciated that by utilizing the diluent pump (249) to aspirate an extracted portion (E) of the pre-diluent solution (P) from the WBC bath (212) and / or utilizing the diluent pump (249) to deliver a transport portion (T) of the extracted portion of the pre-diluent solution (P) to the RBC bath (214), the biological analysis system (210) may be less reliant on the probe (216) to facilitate analysis of the blood sample (B) in the RBC bath (214) as compared to systems that utilize a probe (216) to at least aspirate a portion of the pre-diluent solution (P) from the WBC bath (212) and / or dispense an aspirated portion of the pre-diluent solution (P) to the RBC bath (214). Thus, the probe (216) may be configured to perform various other tasks while the diluent pump (249) aspirates the extract portion (E) of the pre-diluent solution (P) from the WBC bath (212) and / or while the diluent pump (249) delivers the transport portion (T) of the extract portion (E) of the pre-diluent solution (P) to the RBC bath (214). For example, the probe (216) may be configured to facilitate analysis of various other parameters, such as flow imaging of RBCs, WBCs, and PLTs, while the diluent pump (249) facilitates delivery of the transport portion (T) to the RBC bath (214).

[0085] It will also be appreciated that by removing the RBC bath (214) from the path of the probe (216), the biological analysis system (210) may reduce the complexity and / or footprint of the path of the probe (216) as compared to at least a system that includes an RBC bath (214) along the path of the probe (216), thereby allowing the biological analysis system (210) to have a reduced overall size as compared to at least such systems.

[0086] Although the embodiments herein have described the WBC bath (212) and the RBC bath (214) connected to the CBC drip chamber (221), other embodiments are contemplated that utilize the pumped fluid transport techniques contemplated herein. For example, the WBC bath (212) and the RBC bath (214) can be used in a blood analysis system that utilizes flow imaging, where, for example, the WBC bath (212) and the RBC bath (214) are connected to a flow cell and the contents of each are transported through the flow cell for imaging with a high speed, high resolution camera. The principle of using a pump (249) to facilitate fluid transport between chambers or baths of an analysis system to reduce reliance on the probe (216) can be widely used to reduce system load and increase throughput in a variety of situations.

[0087] It should be noted that other embodiments (e.g., systems and associated methods) utilize a wash pump / syringe pump fluid transport system as contemplated herein. For example, a probe (216) can first deliver the diluent and blood sample to the RBC bath (214), and the sample mixture / pre-diluent solution is then transported to the WBC bath (212) via pump (249). Other embodiments can utilize more than two baths (212, 214), in which case pump (249) is used to transport the blood sample solution (e.g., blood and diluent mixture) between one or more of the more than two baths. For example, rather than integrating an HGB transducer with the WBC bath (212) for hemoglobin measurement purposes, a third bath can be utilized for this purpose.

[0088] D. Second Exemplary Method of Analyzing a Biological Sample

[0089] 6 illustrates another exemplary method (300) of analyzing a biological sample, such as a blood sample (B). Although the method (300) is described as being performed using the biological analysis system (210) described above, it will be appreciated that the method (300) may be performed using any suitable biological analysis system.

[0090] The method (300) begins with step (301), in which a blood sample (B) is dispensed into a first fluid analysis chamber, such as a WBC bath (212). In some embodiments, step (301) may be performed by delivering a predetermined amount of blood sample (B), such as about 10 μL to about 16 μL of blood sample (B), via probe (216) to the WBC bath (212) for mixing with a diluent (D) contained in the WBC bath (212), as described above in connection with FIG. 5A. It will be appreciated that prior to step (301), the method (300) may include aspirating blood sample (B) from a specimen (not shown) via probe (216) in a home position, and then moving probe (216) along a path from the home position to the WBC bath (212) to dispense blood sample (B) into the WBC bath (212). The method (300) may also include returning the probe (216) to a home position from the WBC bath (212) without reaching the RBC bath (214).

[0091] Additionally or alternatively, prior to step (301), method (300) may include forming an air gap (G) in first transport line portion (276a), such as by drawing air into first transport line portion (276a) from the WBC bath (212) via diluent pump (249), as described above in connection with FIG. 5I. Additionally or alternatively, prior to step (301), method (300) may include pumping or otherwise transporting diluent (D) to WBC bath (212), such as by delivering a predetermined amount of diluent (D) to WBC bath (212), such as about 2.143 mL to about 3.429 mL of diluent (D), via diluent pump (249), as described above in connection with FIG. 5J.

[0092] Method (300) proceeds from step (301) to step (302), in which mixed bubbles of air (A) are pumped or otherwise transported to a first fluid analysis chamber, such as WBC bath (212). In some embodiments, step (302) may be performed by delivering a burst of mixed bubbles of air (A) to the WBC bath (212) via mixing pump (247) to promote mixing of the contents of the WBC bath (212) into a homogenous state that defines the pre-diluent solution (P), as described above in connection with FIG.

[0093] Method (300) proceeds from step (302) to step (303), in which an extracted portion (E) of pre-dilution solution (P) is pumped or otherwise aspirated from a first fluid analysis chamber, such as a WBC bath (212). In some embodiments, step (303) may be performed by withdrawing a predetermined amount of pre-diluent solution (P), such as about 200 μL to about 250 μL of pre-diluent solution (P), from the WBC bath (212) via the diluent pump (249) to define an extraction portion (E), and withdrawing an air gap (G) and a buffer region (BU) of the extraction portion (E) into the diluent push / pull line (281) such that the predetermined amount of extraction portion (E), such as about 100 μL to about 120 μL of extraction portion (E), substantially fills the second transport line portion (276b) to define a transport portion (T) of the pre-diluent solution (P), as described above in connection with FIG. 5B. Thus, step (303) can be performed without utilizing probe (216) to aspirate the transport portion (T) of pre-dilution solution (P) from the WBC bath (216), thereby freeing probe (216) to perform other tasks simultaneously with the performance of step (303).

[0094] From step (303), method (300) proceeds to step (304), in which the diluent (D) and the transport portion (T) of the pre-diluent solution (P) are pumped or otherwise transported to a second fluid analysis chamber, such as an RBC bath (214). In some embodiments, step (304) may be performed by delivering the transport portion (T) of the pre-diluent solution (P) and a predetermined amount of diluent (D), such as about 2.804 mL to about 3.364 mL of diluent (D), to the RBC bath (214) via a diluent pump (249), as described above in connection with FIG. 5C. Thus, step (304) may be performed without utilizing a probe (216) to deliver the transport portion (T) of the pre-diluent solution (P) to the RBC bath (214), thereby freeing up the probe (216) to perform other tasks simultaneously with the performance of step (304).

[0095] From step (304), method (300) proceeds to step (305), in which lysis solution (L) is pumped or otherwise transported to a first fluid analysis chamber, such as the WBC bath (212). In some embodiments, step (305) may be performed by delivering a predetermined amount of lysis solution (L) to the WBC bath (212), such as about 337.8 μL to about 575.9 μL of lysis solution (L), via lysis solution pump (242), to mix with the portion of pre-diluent solution (P) remaining in the WBC bath (212), as described above in connection with FIG. 5C. It will be appreciated that step (305) may alternatively be performed prior to or simultaneously with step (304).

[0096] From step (305), method (300) proceeds to step (306), where the mixed bubbles of air (A) are pumped or otherwise transported to a first fluid analysis chamber, such as WBC bath (212). In some embodiments, step (306) may be performed by delivering a burst of mixed bubbles of air (A) to the WBC bath (212) via a mixing pump (247) to promote mixing of the contents of the WBC bath (212) to a homogenous state that defines the final WBC solution (FW), as described above in connection with FIG.

[0097] From step (306), method (300) proceeds to step (307), where the mixed bubbles of air (A) are pumped or otherwise transported to a second fluid analysis chamber, such as RBC bath (214). In some embodiments, step (307) may be performed by delivering a burst of mixed bubbles of air (A) to the RBC bath (214) via mixing pump (247) to promote mixing of the contents of the RBC bath (214) to a homogenous state that defines the Final RBC Solution (FR), as described above in connection with FIG. 5D.

[0098] From step (307), the method (300) proceeds to step (308), where the final RBC solution (FR) of a second fluid analysis chamber, such as the RBC bath (214), is analyzed. In some embodiments, step (308) may occur immediately following completion of step (307). In any event, step (308) may include initiating red blood cell / platelet count flow priming and / or data acquisition from the final RBC solution (FR). Additionally or alternatively, step (308) may include pumping or otherwise conveying diluent (D) from the sweep flow tank (241) to the CBC chamber (221), as described above in connection with FIG. 5E. It will be appreciated that step (307) may occur in parallel with one or more of steps (305, 306).

[0099] From step (308), method (300) proceeds to step (309), in which diluent pump (249) is primed to flush diluent input / output line (275), diluent push line (280), and second, third, and fourth transport line portions (276b, 276c, 276d). In some embodiments, step (309) may be performed by filling diluent pump (249) with a predetermined volume of diluent (D) to flush diluent input / output line (275), diluent push line (280), and second, third, and fourth transport line portions (276b, 276c, 276d), as described above in connection with FIG. 5F.

[0100] From step (309), method (300) proceeds to step (310), in which the RBC bath (214) is flushed, thereby cleaning and / or emptied of any waste fluid (W) contained therein. In some embodiments, step (310) may be performed by conveying diluent (D) to the RBC bath (214) via the wash pump (240) and diluent pump (249) and / or draining the waste fluid (W) and / or diluent (D) from the RBC bath (214) via the waste pump (245), as described above in connection with FIG. 5G. It will be appreciated that at the completion of step (310), the RBC bath (214) is substantially empty.

[0101] In the illustrated example, method (300) also proceeds from step (306) to step (311), where the final WBC solution (FW) in a first fluid analysis chamber, such as the WBC bath (212), is analyzed. In some embodiments, step (311) may occur upon completion of one or more incubation periods, which may begin after the mixed bubbles of air (A) are delivered to the WBC bath (212) during performance of step (306). For example, step (311) may include measuring the amount of hemoglobin present in the final WBC solution (FW) when the hemoglobin incubation period is completed, such as with a hemoglobin transducer in the WBC bath (212). Additionally or alternatively, step (311) may include initiating white blood cell count flow priming and / or data acquisition from the final WBC solution (FW) when the WBC incubation period is completed. It will be appreciated that step (311) may be performed in parallel with one or more of steps (307, 308, 309, 310).

[0102] From step (311), method (300) proceeds to step (312), in which the WBC bath (212) is flushed, thereby cleaning, and / or emptied of any waste fluid (W) contained therein. In some embodiments, step (312) may be performed by conveying diluent (D) to the WBC bath (212) via diluent pump (249) and / or draining the waste fluid (W) and / or diluent (D) from the WBC bath (212) via waste pump (245), as described above in connection with FIG. 5H. It will be appreciated that at the completion of step (312), the WBC bath (212) is substantially empty.

[0103] From step (312), method (300) proceeds to step (313), where an air gap (G) is formed for the next cycle. In some embodiments, step (313) may be performed by drawing air from the WBC bath (212) into the first transport line portion (276a) via diluent pump (249), as described above in connection with FIG. 5I.

[0104] From step (313), method (300) proceeds to step (314), where diluent (D) for the next cycle is pumped or otherwise transported to a first fluid analysis chamber, such as WBC bath (212). In some embodiments, step (314) may be performed by delivering a predetermined amount of diluent (D) to the WBC bath (212), such as about 2.143 mL to about 3.429 mL of diluent (D) via diluent pump (249), as described above in connection with FIG.

[0105] In some embodiments, after steps (313, 314) are completed, the method (300) may return to step (301), where another blood sample (B) is dispensed into a first fluid analysis chamber, such as the WBC bath (212), for the next cycle.

[0106] E. Combination Examples

[0107] The following examples relate to various non-exclusive ways in which the teachings herein may be combined or applied. It should be understood that the following examples are not intended to constrain the scope of claims that may be presented at any time in this application or in a later filing of this application. No disclaimers are intended. The following examples are provided for illustrative purposes only. It is contemplated that the various teachings herein may be configured and applied in numerous other ways. It is also contemplated that some variations may omit certain features referenced in the following examples. Thus, none of the aspects or features referenced below should be considered essential unless expressly indicated otherwise at a later date by the inventors or by a successor in interest to the inventors. If a claim including additional features beyond those referenced below is presented in this application or in a later filing related to this application, those additional features shall not be considered as having been added for any reason related to patentability.

[0108] Example 1

[0109] 1. A biological analytical system comprising: (a) a probe configured to dispense a biological sample into a first chamber; and (b) a pump configured to (i) deliver a first delivery of a diluent to the first chamber for mixing with the biological sample to form a sample mixture, (ii) aspirate a portion of the sample mixture from the first chamber, and (iii) deliver the portion of the sample mixture to a second chamber.

[0110] Example 2

[0111] The biological analytical system of Example 1, wherein the probe is configured to aspirate a biological sample from the biological specimen prior to dispensing the biological sample into the first chamber.

[0112] Example 3

[0113] The biological analytical system of Examples 1 or 2, wherein the pump comprises a syringe pump.

[0114] Example 4

[0115] The biological analytical system of any of Examples 1-3, further comprising a CBC drip chamber, the CBC drip chamber configured to count blood cells in at least one of the first or second chambers.

[0116] Example 5

[0117] The biological analytical system of any of Examples 1-4, wherein the pump is configured to deliver a second delivery of diluent to the second chamber while delivering a portion of the sample mixture to the second chamber.

[0118] Example 6

[0119] The biological analysis system of any one of Examples 1 to 5, further comprising a dissolution fluid pump, the dissolution fluid pump configured to transport the dissolution fluid to the first chamber.

[0120] Example 7

[0121] The biological analytical system of any of Examples 1-6, further comprising a first and a second chamber, the first and second chambers configured to be selectively fluidly coupled to one another.

[0122] Example 8

[0123] The biological analytical system of Example 7 further comprises a transport line configured to selectively fluidly couple the first and second chambers with one another, and the pump configured to aspirate a portion of the sample mixture into the transport line prior to conveying the portion of the sample mixture through the transport line to the second chamber.

[0124] Example 9

[0125] The biological analytical system of Example 8, further comprising at least one diluent push / pull line configured to be selectively fluidly coupled to the transport line, wherein the pump is configured to draw an air gap in the at least one diluent push / pull line while aspirating a portion of the sample mixture into the transport line.

[0126] Example 10

[0127] 1. A biological analytical system comprising: (a) a first chamber; (b) a second chamber; (c) a probe configured to dispense a biological sample into the first chamber; (d) a first pump configured to transport a portion of the biological sample from the first chamber to the second chamber; and (e) a second pump configured to deliver a fluid medium to at least one of the first chamber or the second chamber.

[0128] Example 11

[0129] The biological analytical system of Example 10, wherein the fluid medium comprises a lysis solution, and the second pump is configured to transport the lysis solution to the first chamber after transporting a portion of the biological sample to the second chamber.

[0130] Example 12

[0131] The biological analytical system of Example 10 or 11, further comprising a CBC drip chamber, the CBC drip chamber fluidly coupled to at least one of the first or second chambers for counting blood cells in at least one of the first or second chambers.

[0132] Example 13

[0133] The biological analytical system of any of Examples 10-12, wherein the first pump is configured to deliver a first delivery of a diluent to the first chamber for mixing with the biological sample to generate a sample mixture prior to transporting a portion of the biological sample to the second chamber.

[0134] Example 14

[0135] The biological analytical system of Example 13, wherein the first pump is configured to deliver a second delivery of diluent to the second chamber while transporting a portion of the biological sample to the second chamber.

[0136] Example 15

[0137] The biological analytical system of any of Examples 10-14, wherein the first and second chambers are configured to be selectively fluidly coupled to one another.

[0138] Example 16

[0139] The biological analytical system of Example 15 further comprises: (a) a transport line configured to selectively fluidly couple the first and second chambers with one another; and (b) at least one diluent push / pull line configured to be selectively fluidly coupled to the transport line, wherein the first pump is configured to draw an air gap in the at least one diluent push / pull line while aspirating a portion of the biological sample into the transport line.

[0140] Example 17

[0141] The biological analysis system of any of Examples 10-16, wherein the first chamber comprises a hemoglobin transducer.

[0142] Example 18

[0143] The biological analysis system of Example 17, wherein the hemoglobin transducer comprises a filtered light source and an optical sensor.

[0144] Example 19

[0145] 1. A method for analyzing a biological sample, comprising: (a) conveying a diluent to a first chamber via a first pump; (b) dispensing a blood sample via a probe into the first chamber to generate a sample mixture; and (c) conveying a portion of the sample mixture from the first chamber to a second chamber via the first pump.

[0146] Example 20

[0147] The method of Example 19, further comprising conveying the lysis solution to the first chamber via a second pump.

[0148] Example 21

[0149] The biological analytical system of Example 8 further comprising a pair of transport line valves positioned along the transport line, the pair of transport line valves separated from one another by a transport line portion having a fixed volume, the transport line portion configured to receive a portion of the sample mixture having a predetermined volume substantially equal to the fixed volume of the transport line portion.

[0150] Example 22

[0151] The biological analytical system of Example 21, wherein each transfer line valve of the pair of transfer line valves comprises a three-way valve.

[0152] Example 23

[0153] The biological analytical system of Example 9, wherein the at least one diluent push / pull line comprises a pair of diluent push / pull lines.

[0154] Example 24

[0155] The biological analytical system of Example 23, further comprising a pair of three-way valves positioned along the transport line, each three-way valve of the pair of three-way valves coupled to a respective diluent push / pull line of the pair of diluent push / pull lines.

[0156] Example 25

[0157] The biological analytical system of Example 24, wherein the pairs of three-way valves are separated from each other by transfer line segments, the transfer line segments having fixed volumes.

[0158] Different configurations of the components depicted in the figures or described above, as well as components and steps not shown or described, are possible. Similarly, some features and subcombinations are useful and may be employed without reference to other features and subcombinations. The embodiments of the invention have been described for purposes of illustration and not of limitation. Alternative embodiments will also become apparent to the reader of this patent. In some cases, method steps or operations may be performed or executed in a different order, or operations may be added, deleted, or modified. It can be recognized that in certain aspects of the invention, a single component may be replaced by multiple components, and multiple components may be replaced by a single component, to provide an element or structure or to perform a given function or functions. Except where such a substitution is not operable to implement a particular embodiment of the invention, such substitution is considered to be within the scope of the invention. Thus, the invention is not limited to the embodiments described above or depicted in the figures, and various embodiments and variations can be made without departing from the scope of the following claims. [Explanation of symbols]

[0159] 10 Biological Analysis Systems 12 First bath, first fluid analysis chamber, WBC bath 14 Second bath, second fluid analysis chamber, RBC bath 16 Probes 18 Complete blood count (CBC) box 20 Hemoglobin Transducer 30 Diluent storage section 32 Dissolving solution storage section 40 Cleaning Pump 42 Dissolving Liquid Pump 44 Wastewater Pump 46 CBC waste container 50 Diluent supply line 52 Filling and flushing pump valve 54 Common RBC / WBC input line 56 First T-junction 58 Second T-junction 60 RBC input lines 62 Washing RBC valve 64 1st WBC input line 66 Washing WBC valve 68 Dissolving solution supply line 70 Dissolving Liquid Valve 72 Second WBC Input Line 74 RBC drainage line 76 WBC drainage line 78 Third T-junction 80 Common RBC / WBC drainage line 82, 84 RBC and WBC drainage valves 210 Biological Analysis Systems 212 First bath, first fluid analysis chamber, WBC bath 214 Second bath, second fluid analysis chamber, RBC bath 216 Sample Probe 221 CBC Drip Chamber 222 Air Pressure Transducer 230 Diluent storage section 232 Dissolving liquid storage section 233 Cleaning agent storage section 241 Sweep Flow Tank 242 Dissolving Liquid Pump 244 Vacuum Pump 245 Wastewater Pump 246 Waste liquid container 247 Mixing Pump 248 Atmosphere 249 Diluent Pump 250 Diluent supply line 251 First Multi-Flow Unit 252 Diluent / Detergent Valve 254 Cleaning agent supply line 255 Detergent valve 256 RBC input lines 257 Washing RBC valve 258 Sweep flow input line 259 Cleaning Sweep Flow Valve 260 Second Multi-Flow Unit Input Line 261 Second Multi-Flow Unit 264 WBC input line 265 Washing WBC valve 266 Diluent input line 267 Diluent input valve 268 Common Sweep Flow Input Line 269 ​​First Y-branch 270 Sweep Flow Valve 271, 272 Sweep flow line 273, 274 Sweep flow valve 275 Diluent Input / Output Lines 276a, 276b, 276c, 276d Transport line section 277, 278, 279 Transport line valves 280 Diluent Push Line 281 Diluent Push / Pull Line 282 Dissolving solution supply line 283 RBC drainage line 284 Third Multi-Flow Unit 285 RBC drainage valve 286 Mixed Air Line 288 Mixing Pump Valve 289 WBC drainage line 290 WBC drainage valve 291 Mixed Air Line 292 Third Y-branch 293 CBC drainage line 294 CBC drainage valve 295 Wastewater Line 297 Wastewater Line 298 Vacuum Line 299 Vacuum Line Valve D Diluent L solution B. Blood sample W Waste fluid C Damage area T Transport part E extraction part P predilution solution FW Final WBC solution G Air gap

Claims

1. 1. A biological analytical system comprising: (a) a probe configured to dispense a biological sample into a first chamber; (b) a pump, (i) delivering a first delivery of a diluent to the first chamber for mixing with the biological sample to form a sample mixture; (ii) aspirating a portion of the sample mixture from the first chamber; (iii) conveying the portion of the sample mixture to a second chamber. and a pump configured to:

2. The biological analytical system of claim 1 , wherein the probe is configured to aspirate the biological sample from a biological specimen prior to dispensing the biological sample into the first chamber.

3. The biological analytical system of claim 1 , wherein the pump comprises a syringe pump.

4. 10. The biological analytical system of claim 1, further comprising a CBC drip chamber configured to count blood cells in at least one of the first or second chambers.

5. 2. The biological analytical system of claim 1, wherein the pump is configured to deliver a second delivery of diluent to the second chamber while delivering the portion of the sample mixture to the second chamber.

6. The biological analytical system of claim 1 , further comprising a lysate pump, the lysate pump configured to deliver lysate to the first chamber.

7. The biological analytical system of claim 1 , further comprising the first and second chambers, the first and second chambers configured to be selectively fluidly coupled to one another.

8. 8. The biological analytical system of claim 7, further comprising a transport line configured to selectively fluidly couple the first and second chambers with one another, the pump configured to aspirate the portion of the sample mixture into the transport line prior to transporting the portion of the sample mixture through the transport line to the second chamber.

9. 10. The biological analytical system of claim 8, further comprising at least one diluent push / pull line configured to be selectively fluidly coupled to the transport line, the pump configured to draw an air gap in the at least one diluent push / pull line while aspirating the portion of the sample mixture into the transport line.

10. 1. A biological analytical system comprising: (a) a first chamber; (b) a second chamber; and (c) a probe configured to dispense a biological sample into the first chamber; (d) a first pump configured to transport a portion of the biological sample from the first chamber to the second chamber; and (e) a second pump configured to deliver a fluid medium to at least one of the first chamber or the second chamber.

11. 11. The biological analytical system of claim 10, wherein the fluid medium comprises a lysis liquid, and the second pump is configured to deliver the lysis liquid to the first chamber after transporting the portion of the biological sample to the second chamber.

12. 11. The biological analytical system of claim 10, further comprising a CBC drip chamber, said CBC drip chamber fluidly coupled to at least one of said first or second chambers for counting blood cells in said at least one of said first or second chambers.

13. 11. The biological analytical system of claim 10, wherein the first pump is configured to deliver a first delivery of a diluent to the first chamber for mixing with the biological sample to form a sample mixture prior to transporting the portion of the biological sample to the second chamber.

14. 14. The biological analytical system of claim 13, wherein the first pump is configured to deliver a second delivery of diluent to the second chamber while transporting the portion of the biological sample to the second chamber.

15. The biological analytical system of claim 10 , wherein the first and second chambers are configured to be selectively fluidly coupled to one another.

16. (a) a transfer line configured to selectively fluidly couple the first and second chambers with one another; (b) at least one diluent push / pull line configured to be selectively fluidly coupled to the transfer line; 16. The biological analytical system of claim 15, wherein the first pump is configured to draw an air gap in the at least one diluent push / pull line while aspirating the portion of the biological sample into the transport line.

17. The biological analytical system of claim 10 , wherein the first chamber comprises a hemoglobin transducer.

18. 20. The biological analytical system of claim 17, wherein the hemoglobin transducer comprises a filtered light source and an optical sensor.

19. 1. A method for analyzing a biological sample, comprising: (a) conveying a diluent to a first chamber via a first pump; (b) dispensing a blood sample via a probe into the first chamber to generate a sample mixture; (c) transporting a portion of the sample mixture from the first chamber to a second chamber via the first pump.

20. 20. The method of claim 19, further comprising delivering a lysis liquid to the first chamber via a second pump.

21. 10. The biological analytical system of claim 8, further comprising a pair of transport line valves located along the transport line, the pair of transport line valves separated from one another by a transport line portion having a fixed volume, the transport line portion configured to receive the portion of the sample mixture having a predetermined volume substantially equal to the fixed volume of the transport line portion.

22. 22. The biological analytical system of claim 21, wherein each transfer line valve of the pair of transfer line valves comprises a three-way valve.

23. 10. The biological analytical system of claim 9, wherein the at least one diluent push / pull line comprises a pair of diluent push / pull lines.

24. 24. The biological analytical system of claim 23, further comprising a pair of three-way valves located along the transport line, each three-way valve of the pair of three-way valves coupled to a respective diluent push / pull line of the pair of diluent push / pull lines.

25. 25. The biological analytical system of claim 24, wherein the pairs of three-way valves are separated from one another by transfer line segments, the transfer line segments having a fixed volume.

Images