Automated sample stream detection and validation

Abstract

When operating a biological analyzer, a relative position between a camera and a flowcell may be set and confirmed using a method which sequentially focuses to a fixed target and a flow of particles through the flowcell. Such a method may include setting a first relative position by focusing to the target, then setting a second relative position by focusing to the flow of particles. This second relative position may then be confirmed by analyzing images of second particles passing through the flowcell.

Description

AUTOMATED SAMPLE STREAM DETECTION AND VALIDATIONCROSS REFERENCE TO RELATED APPLICATIONS

[0001] This claims priority from provisional application 63 / 693,122, filed at the United States Patent Office on September 10, 2025, and entitled “Automated Sample Stream Detection and Validation,” the disclosure of which is hereby incorporated in its entirety.BACKGROUND

[0002] Biological analyzers utilizing imaging can be used, for instance, in the urine analysis and blood analysis space. The technology involves passing a biological sample (e.g., blood or urine) through a flowcell of a biological analyzer, and utilizing an image capture device to capture images of the relevant biological material (e.g., cells or particles). The cells or particles are then classified or categorized, and can be used to assess a patient’s health. Proper image focal quality is important to ensure the biological material (e.g., cells or particles) is clearly imaged and can be categorized correctly (e.g., through a computerized classifier).

[0003] Several parameters can impact proper image focal quality, including manufacturing variances with various flowcells (e.g., impacting an ideal focal position), environmental phenomena such as temperature, or natural variations that can occur as a machine is operated.

[0004] There is a need for systems, methods, and devices capable of determining proper image focal qualities in a biological analysis capacity, as well as being able to take an action to ensure a proper image focal quality (e.g., adjusting one or more of an image capture device, objective, and / or flowcell).BRIEF SUMMARY

[0005] The present disclosure relates to systems and methods for operating imaging biological analyzers. Methods which may be implemented based on this disclosure may comprise setting- 1 -01337880810419 4919-0466-2881v3a first relative position between a camera and a flowcell by focusing on a target, where the target has a fixed position relative to the flowcell. Such a method may also comprise, after setting the first relative position, setting a second relative position between the camera and the flowcell by focusing to first particles passing through the flowcell. Additionally, in such a method, after the second relative position has been set, it may be confirmed by analyzing images of second particles passing through the flowcell.

[0006] The disclosed technology may also be implemented in other manners, such as in the form of systems or computer readable media programmed to performed methods such as described above. Accordingly, the above description of a method which may be implemented based on this disclosure should be understood as being illustrative only, and should not be treated as limiting.BRIEF DESCRIPTION OF THE DRAWINGS

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

[0008] FIG. l is a schematic illustration, partly in section and not to scale, showing operational aspects of an exemplary flowcell which may be used in an analyzer configured to capture and analyze images.

[0009] FIG. 2 illustrates a method to focus an imaging device on a location in a flowcell through which a sample stream will transport particles to be imaged.

[0010] FIG. 3 illustrates a potential displacement of a sample stream relative to an expected position.

[0011] FIG. 4 illustrates a method which includes setting a rough relative position between setting first and second relative positions.- 2 -01337880810419 4919-0466-2881v3

[0012] FIG. 5 illustrates a graphical representation of focus indicator values.

[0013] The drawings are not intended to be limiting in any way, and it is contemplated that various embodiments of the invention may be carried out in a variety of other ways, including those not necessarily depicted in the drawings. The accompanying drawings incorporated in and forming a part of the specification illustrate several aspects of the present invention, and together with the description serve to explain the principles of the invention; it being understood, however, that this invention is not limited to the precise arrangements shown.DETAILED DESCRIPTION

[0014] The present disclosure relates to operating imaging biological analyzers while identifying a proper focal position. In various examples, several scenarios can contribute to an offset between an actual and ideal relative imaging position (e.g., the relative imaging position between one or more of a camera, objective, and flowcell). For instance, manufacturing variances in flowcells can contribute to minute irregularities in flowcell design which can impact sample focal quality within an imaging region of the flowcell. Environmental factors such as humidity or temperature can also contribute, as can various variations that can occur as a biological analysis machine is run over the course of an extended time period.

[0015] In one example, the biological analyzer utilizes flow imaging principles (that is flowing a blood sample past an imaging device). An example of the type of instrument which may be used with this technology is shown in FIG. 1, which shows an exemplary flowcell 22 which may be used in an analyzer for conveying a sample fluid through a viewing zone 23 of a high optical resolution imaging device 24 (e.g., a camera) in a configuration for imaging microscopic particles in a sample flow stream 32 using digital image processing. Flowcell 22 is coupled to a source 25 of sample fluid which may have been subjected to processing, such as contact with a particle contrast agent composition and heating. Flowcell 22 is also coupled to one or more sources 27 of a particle and / or intracellular organelle alignment liquid (PIOAL) / sheath fluid,- 3 -01337880810419 4919-0466-2881v3such as a clear glycerol solution having a viscosity that is greater than the viscosity of the sample fluid, an example of which is disclosed in U.S. Pat. Nos. 9,316,635 and 10,451,612, the disclosures of which are hereby incorporated by reference in their entirety. In various examples, the sample fluid can be a blood sample which is imaged to detect blood particles (e g., various combinations of red blood cells, platelets, and white blood cells including monocytes, basophils, neutrophils, eosinophils, and lymphocytes), or a urine sample which is imaged to detect urine particles (e.g., various combinations of red and white blood cells, casts, bacteria, sperm).

[0016] The sample fluid is injected through a flattened opening at a distal end 28 of a sample feed tube 29, and into the interior of the flowcell 22 at a point where the PIOAL flow has been substantially established resulting in a stable and symmetric laminar flow of the PIOAL above and below (or on opposing sides of) the ribbon-shaped sample stream. The sample and PIOAL streams may be supplied by precision metering pumps that move the PIOAL with the injected sample fluid along a flowpath that narrows substantially. The PIOAL envelopes and compresses the sample fluid in the zone 21 where the flowpath narrows. Hence, the decrease in flowpath thickness at zone 21 can contribute to a geometric focusing of the sample flow stream 32. The sample flow stream 32 is enveloped and carried along with the PIOAL downstream of the narrowing zone 21, passing in front of, or otherwise through the viewing zone 23 of, the high optical resolution imaging device 24 where images are collected, for example, using a CCD 48 observing the sample stream through a viewing port 57. Processor 18 can receive, as input, pixel data from CCD 48. The sample fluid ribbon flows together with the PIOAL to a discharge 33.

[0017] As shown here, the narrowing zone 21 can have a proximal flowpath portion 21a having a proximal thickness PT and a distal flowpath portion 21b having a distal thickness DT, such that distal thickness DT is less than proximal thickness PT. The sample fluid can therefore be injected through the distal end 28 of sample tube 29 at a location that is distal to the proximal- 4 -01337880810419 4919-0466-2881v3portion 21a and proximal to the distal portion 21b. Hence, the sample fluid can enter the PIO AL envelope as the PIO AL stream is compressed by the zone 21, wherein the sample fluid injection tube has a distal exit port through which sample fluid is injected into flowing sheath fluid, the distal exit port bounded by the decrease in flowpath size of the flowcell.

[0018] The digital high optical resolution imaging device 24 with objective lens 46 is directed along an optical axis that intersects the ribbon-shaped sample flow stream 32. The relative distance between the objective 46 and the flowcell 33 is variable by operation of a motor drive 54, for resolving and collecting a focused digitized image on a photosensor array. Additional information regarding the construction and operation of an exemplary flowcell such as shown in FIG. 1 is provided in U.S. Patent 9,322,752, entitled “Flowcell Systems and Methods for Particle Analysis in Blood Samples,” filed on March 17, 2014, the disclosure of which is hereby incorporated by reference in its entirety.

[0019] To facilitate the operation of a biological analyzer which performs imaging using a flowcell system such as shown in FIG. 1, the disclosed technology may be used to implement a method such as shown in FIG. 2 to focus the system’s imaging device on a location in the flowcell 22 through which a sample stream will transport particles to be imaged. As shown in FIG. 2, such a method may begin with setting 201 a first relative position between the camera and the flowcell 22. This relative position may be one where the imaging device 24 is focused on focal plane that includes an axis along which the flow stream is expected to travel (e.g., the y-axis of FIG. 3)- that is, a focal plane perpendicular to the optical axis 50 of the camera. As discussed below, this first relative position may be treated as a baseline for further adjustment steps, and may be done by, for example, focusing on a target which has a fixed position relative to the flowcell. The target can be a high contrast figure defining a known location along the optical axis of the imaging device 24, and may have a known displacement distance relative to the location of the ribbon-shaped sample stream. A contrast measurement algorithm can be employed specifically on the target. In one example, the position of the imaging device 24 can- 5 -01337880810419 4919-0466-2881v3be varied along a line parallel to the optical axis of the high optical resolution imaging device or the digital image capture device, to find the depth or distance at which one or more maximum differential amplitudes are found among the pixel luminance values occurring along a line of pixels in the image that is known to cross over an edge of the target. In some cases, the target has no variation along the line parallel to the optical axis, which is also the line along which a motorized control operates to adjust the position of the high optical resolution imaging device 24 to provide the recorded displacement distance. Having found the location of optimal or desired focus on the target, the relative positions of the high optical resolution imaging device objective 24 and the flowcell 22 can be displaced by the recorded displacement distance to provide the optimal or desired focus position for particles in the ribbon-shaped sample stream 32. Additional information on approaches that may be used in setting 201 the first relative position is provided in U.S. Patent 9,857,361 entitled “Flowcell, Sheath fluid, and Autofocus Systems and Methods for Particle Analysis in Urine Samples” and U.S. Patent 10,705,008 entitled “Autofocus Systems and Methods for Particle Analysis in Blood Samples,” each of which is hereby incorporated by reference in its entirety.

[0020] The first relative position adjustment comprises movement of a flowcell relative to an imaging device or camera. In various embodiments, this can comprise adjusting any one or more of a camera, objective (e.g., microscope), and flowcell. For example, a camera can be adjusted relative to a stationary flowcell, a flowcell can be adjusted relative to a stationary camera, or a camera connected to an objective can be adjusted relative to a stationary flowcell.

[0021] Continuing with the discussion of FIG. 2, after the first relative position has been set 201, a second relative position may be set 202 to allow for a more precise focal adjustment (e.g., adjustment relative to particles flowing through the flowcell). An example of this type of displacement is shown in FIG. 3, in which the sample stream 32 is displaced relative to the expected position along the optical axis (shows as the Z axis in FIG. 3). This may be done by flowing a control material through the flowcell and focusing on particles carried by the sample- 6 -01337880810419 4919-0466-2881v3stream. These control particles may be composed of various materials - natural biological material, such as human or animal derived blood cells, or synthetic material such as metallic or polymeric beads or other synthetic particles, depending on the control material being used in a particular case. It is also possible that a setting 202 the second relative position may be performed by focusing on particles in a patient sample, rather than particles in a control material, in which case the particles may be particles in a blood or urine sample, or other type of body fluid sample which may be processed by the analyzer in question. In the context of FIG. 3, the flow is shown in a bottom-to-top manner, though in other configurations this can be flipped (e.g., in a top-to-bottom manner).

[0022] Whatever kinds of materials are used in setting 202 the second relative position, the image(s) of that material can be used to calculate focus values for setting 202 the position. For example, in some cases, focus values may be calculated as focus indicator values determined by using a fast Fourier transform (FFT) to convert images captured by the camera from the spatial to the frequency domain, then applying a high pass filter and averaging the magnitude of the FFT weighted by the distance from the center of origin (e.g., center of the flowcell). Focus values may also be calculated in other ways, such as by using contrast to measure focus as described above, or using ring-based pixel binning, as described in international application PCT / US2023 / 011759 entitled “measure image quality of blood cell images,” the disclosure of which is hereby incorporated by reference in its entirety. Whatever types of calculations are used, the focus values can be applied to set 202 the second relative position, such as by treating the lowest focus indicator value as the value indicating the best focus. The position of the camera corresponding to that lowest focus indicator value can be set 202 as the second relative position in the method of FIG. 2. An illustration of a curve showing potential focus indicator values is provided in FIG. 5, with the second relative position which may be set 202 in a method such as shown in FIG. 2 corresponding to the lowest point in the curve.- 7 -01337880810419 4919-0466-2881v3

[0023] It is also possible that a multi-phased approach may be used when setting 202 the second relative position. This may be done, for example, as shown in FIG. 4, which illustrates setting 202 the second relative position by first setting 401 a rough relative position, and then setting 402 a fine relative position. In this type of implementation, setting 401 the rough relative position may comprise relative movement between the camera and the flowcell. In various embodiments, this can comprise adjusting any one or more of a camera, objective (e.g., microscope), and flowcell. For example, a camera can be adjusted relative to a stationary flowcell, a flowcell can be adjusted relative to a stationary camera, or a camera connected to an objective can be adjusted relative to a stationary flowcell. The adjustment can be performed at a rough adjustment speed (e.g., 4 pm / s, or some other speed from 2 pm / s to 6 pm / s; 8 pm / s, or some other speed between 6 pm / s and 10 pm / s; 12 pm / s, or some other speed between 10 pm / s and 16 pm / s, etc.) through a rough adjustment range (e.g., a span of between 40 and 80 pm, such as about a 60 pm span from about 30 pm closer to the camera from the first relative position to about 30 pm closer to the flowcell from the first relative position) while a set of particles (e.g., control particles from a control material) was flowing through the flowcell. Focus values could then be calculated for the various positions of the camera through the rough adjustment range and used to identify the best focus from the rough adjustment range in a manner similar to that described above in the context of FIG. 2.

[0024] With a rough relative position having been set 401, that rough relative position may be used in setting 202 the fine relative position. This may be done, for example, by relative movement between the camera and the flowcell. In various embodiments, this can comprise adjusting any one or more of a camera, obj ective (e.g., microscope), and flowcell. For example, a camera can be adjusted relative to a stationary flowcell, a flowcell can be adjusted relative to a stationary camera, or a camera connected to an objective can be adjusted relative to a stationary flowcell. The adjustment can be performed through a fine adjustment range (e.g., a span of between 8 and 16 pm, such as about a 12 pm span from about 6 pm closer to the camera from the rough relative position to about 6 pm closer to the flowcell from the rough relative position)- 8 -01337880810419 4919-0466-2881v3at a fine adjustment speed (e.g.,0.4 pm / s, or some other speed from 0.2 pm / s to 0.6 pm / s; 0.8 pm / s, or some other speed between -.6 pm / s and 1 pm / s; 12 pm / s, or some other speed between 1.0 pm / s and 1.6 pm / s, etc.) while the biological material flows through the flowcell, and using images captured by the camera to calculate focus values as described above. In this way, once the fine focus value had been determined, it could be treated as the second relative position, thereby allowing implementations of the disclosed technology to quickly and with high precision identify a proper focal position for a camera.

[0025] In the method of FIG. 2, after the second relative position has been set 202, that position may be confirmed 203, either validating that the analyzer was suitable for processing patient samples (if the confirmation was positive) or to trigger some follow up action such as redetermining the relative position or generating an alarm for an operator (if the confirmation was negative). This confirmation may be performed by establishing a second ribbon shaped stream through the flowcell, and analyzing it as it is subjected to the same kind of handling as would be applied to patient samples during the analyzer’ s normal operation. The second ribbon shaped stream may be a second stream of the same material used in setting 202 the second relative position. For example, if a control material was used in setting 202 the second relative position, then, to confirm 203 that position, a second aspiration of that same control material may be flowed through the flowcell. Alternatively, a different material may be used for confirming 203 the second relative position than was used to set 202 it. For example, if the second relative position was set 202 using a control material, then it may be confirmed 203 using a different control material, or using a patient sample.

[0026] Whatever material is flowed through the flowcell during the confirmation 203, the analysis performed based on that material may include calculating focus values (e.g., focus indicator values such as described previously) for imaged particles in the second biological material and then confirming that those values satisfied one or more acceptability criteria. For example, in some cases, a mean may be calculated using the focus values of the imaged particles, and- 9 -01337880810419 4919-0466-2881v3confirming 203 the suitability of the second relative position may be performed by confirming that that mean is within a passing range (e.g., in a case where lower values indicate better focus, confirming that the mean is below an acceptability threshold). As another example, in some cases, the uniformity of the distribution of focus values (e.g., as calculated using deviations between minimum and maximum values) throughout the field of view of the camera may be evaluated, with the second relative position being confirmed 203 as being acceptable if the distribution was sufficiently uniform (e.g., deviation was below an acceptability threshold). Other approaches to confirming 203 the second relative position are also possible, and may be used in some implementations. For example, in some cases, confirmation 203 may involve a combination of parameters, such as in a case where the second relative position would be confirmed as acceptable if the mean focus value was below a first acceptability threshold, and the distribution of focus values was sufficiently regular. Accordingly, the exemplary approaches described above should be understood as being illustrative only, and should not be treated as limiting.

[0027] As a further illustration of potential implementations and applications of the disclosed technology, the following examples are provided of non-exhaustive ways in which the teachings herein may be combined or applied. It should be understood that the following examples are not intended to restrict the coverage of any claims that may be presented at any time in this application or in subsequent filings of this application. No disclaimer is intended. The following examples are being provided for nothing more than merely illustrative purposes. It is contemplated that the various teachings herein may be arranged and applied in numerous other ways. It is also contemplated that some variations may omit certain features referred to in the below examples. Therefore, none of the aspects or features referred to below should be deemed critical unless otherwise explicitly indicated as such at a later date by the inventors or by a successor in interest to the inventors. If any claims are presented in this application or in subsequent filings related to this application that include additional features beyond those- 10 -01337880810419 4919-0466-2881v3referred to below, those additional features shall not be presumed to have been added for any reason relating to patentability.

[0028] Example 1

[0029] A method of operating a biological analyzer comprising: setting a first relative position between a camera and a flowcell by focusing to a target, the target having a fixed position relative to the flowcell; after setting the first relative position setting a second relative position between the camera and the flowcell based on focusing to first particles passing through the flowcell; and confirming the second relative position based on capturing and analyzing images of second particles passing through the flowcell.

[0030] Example 2

[0031] The method of example 1, wherein setting the second relative position comprises moving the camera along an optical axis of the camera.

[0032] Example 3

[0033] The method of an of examples 1-2, wherein: setting the second relative position comprises: setting a rough relative position based on focusing to the first particles passing through the flowcell; and setting a fine relative position based on the rough relative position and on focusing to third particles passing through the flowcell; and the second relative position is the fine relative position.

[0034] Example 4

[0035] The method of example 3, wherein: setting the rough relative position comprises moving the camera along an optical axis of the camera at a rough adjustment speed through a rough adjustment range; and setting the fine relative position comprises moving the camera along the optical axis of the camera at a second adjustment speed through a second adjustment range.- 11 -01337880810419 4919-0466-2881v3

[0036] Example 5

[0037] The method of example 4, wherein the second adjustment speed is slower than the rough adjustment speed.

[0038] Example 6

[0039] The method of any of examples 4-5, wherein the second adjustment range is smaller than the rough adjustment range.

[0040] Example 7

[0041] The method of any of examples 4-6, wherein rough adjustment range is greater than 50 micrometers.

[0042] Example 8

[0043] The method of any of examples 4-7, wherein the second adjustment range is under 20 micrometers.

[0044] Example 9

[0045] The method of any of examples 4-8, wherein the rough adjustment range is centered at the first relative position.

[0046] Example 10

[0047] The method of any of examples 4-9, wherein the second adjustment range is centered at the rough relative position.

[0048] Example 11- 12 -01337880810419 4919-0466-2881v3

[0049] The method of any of examples 1-10, wherein: the first particles are particles from a first aspiration of a blood quality control material; and the second particles are particles from a second aspiration of the blood quality control material.

[0050] Example 12

[0051] The method of any of examples 1-10, wherein the first particles are particles comprised by a first quality control sample and the second particles are particles comprised by a second quality control sample.

[0052] Example 13

[0053] The method of any of examples 1-12, wherein the biological analyzer is a urine analysis instrument.

[0054] Examples 14

[0055] The method of any of examples 1-13, wherein the flowcell comprises a flow passage configured to convey the first and second particles, and wherein the target is located on the flowcell and externally of the flow passage.

[0056] Example 15

[0057] A biological analysis system comprising: at least one processor; and a non-transitory computer readable medium having stored thereon instruction which, when executed by the at least one processor, cause the system to perform the method of any of examples 1-14.

[0058] Example 16

[0059] A biological analysis system comprising: a camera, a flowcell, and a target wherein the target has a fixed position relative to the flowcell; at least one processor; a computer-readable medium storing instructions that, when executed by the at least one processor, perform a- 13 -01337880810419 4919-0466-2881v3method comprising: setting a first relative position between a camera and a flowcell by focusing to the target, the target having a fixed position relative to the flowcell; after setting the first relative position, setting a second relative position between the camera and the flowcell based on focusing to first particles passing through the flowcell; and confirming the second relative position based on capturing and analyzing images of second particles passing through the flowcell.

[0060] Example 17

[0061] The biological analysis system of example 16 wherein setting the second relative position comprises moving the camera along an optical axis of the camera.

[0062] Example 18

[0063] The biological analysis system of any of examples 16-17, wherein: setting the second relative position comprises: setting a rough relative position based on focusing to the first particles passing through the flowcell; and setting a fine relative position based on the rough relative position and on focusing to third particles passing through the flowcell; and the second relative position is the fine relative position.

[0064] Example 19

[0065] The biological analysis system of example 18, wherein: setting the rough relative position comprises moving the camera along an optical axis of the camera at a rough adjustment speed through a rough adjustment range; and setting the fine relative position comprises moving the camera along the optical axis of the camera at a second adjustment speed through a second adjustment range.

[0066] Example 20- 14 -01337880810419 4919-0466-2881v3

[0067] The biological analysis system of example 19, wherein the second adjustment speed is slower than the rough adjustment speed.

[0068] Example 21

[0069] The biological analysis system of any of examples 19-20, wherein the second adjustment range is smaller than the rough adjustment range.

[0070] Example 22

[0071] The biological analysis system of any of examples 19-21, wherein rough adjustment range is greater than 50 micrometers.

[0072] Example 23

[0073] The biological analysis system of any of examples 19-22, wherein the second adjustment range is under 20 micrometers.

[0074] Example 24

[0075] The biological analysis system of any of examples 19-23, wherein the rough adjustment range is centered at a center of the flowcell.

[0076] Example 25

[0077] The biological analysis system of any of examples 19-24, wherein the second adjustment range is centered at the rough relative position.

[0078] Example 26

[0079] The biological analysis system of any of examples 16-25, wherein: the first particles are particles from a first aspiration of a blood quality control material; and the second particles are particles from a second aspiration of the blood quality control material.- 15 -01337880810419 4919-0466-2881v3

[0080] Example 27

[0081] The biological analysis system of any of examples 16-25, wherein the first particles are particles comprised by a first quality control sample and the second particles are particles comprised by a second quality control sample.

[0082] Example 28

[0083] The biological analysis system of any of examples 16-27, wherein the biological analyzer is a urine analysis instrument.

[0084] Example 29

[0085] The biological analysis system of any of examples 16-28, wherein the flowcell comprises a flow passage configured to convey the first and second biological materials, and wherein the target is located on the flowcell and externally of the flow passage.

[0086] Example 30

[0087] A method comprising performing the set of acts the instructions stored on the one or more non- transitory computer readable mediums of any of examples 16-29 are to perform when executed.

[0088] Example 31

[0089] An analyzer, comprising: a camera; a flowcell; a target wherein the target has a fixed position relative to the flowcell; at least one processor; and a computer-readable medium storing instructions that, when executed by the at least one processor, perform a method comprising: setting a first relative position between a camera and a flowcell by focusing to the target; after setting the first relative position, setting a second relative position between the camera and the flowcell based on focusing to first particles passing through the flowcell; and confirming the- 16 -01337880810419 4919-0466-2881v3second relative position based on capturing and analyzing images of second particles passing through the flowcell.

[0090] Example 31

[0091] The analyzer of example 30, wherein setting the second relative position comprises moving the camera along an optical axis of the camera.

[0092] Example 32

[0093] The analyzer of example 30, wherein: setting the second relative position comprises: setting a rough relative position based on focusing to the first particles passing through the flowcell; and setting a fine relative position based on the rough relative position and on focusing to third particles passing through the flowcell; and the second relative position is the fine relative position.

[0094] Example 33

[0095] The analyzer of example 32, wherein: setting the rough relative position comprises moving the camera along an optical axis of the camera at a rough adjustment speed through a rough adjustment range; and setting the fine relative position comprises moving the camera along the optical axis of the camera at a second adjustment speed through a second adjustment range.

[0096] Example 34

[0097] The analyzer of example 33, wherein the second adjustment speed is slower than the rough adjustment speed.

[0098] Example 35

[0099] The analyzer of example 33, wherein the second adjustment range is smaller than the rough adjustment range.- 17 -01337880810419 4919-0466-2881v3

[0100] Example 36

[0101] The analyzer of example 33, wherein rough adjustment range is greater than 50 micrometers.

[0102] Example 37

[0103] The analyzer of example 33, wherein the second adjustment range is under 20 micrometers.

[0104] Example 38

[0105] The analyzer of example 33, wherein the rough adjustment range is centered at the first relative position

[0106] Example 39

[0107] The analyzer of example 33, wherein the second adjustment range is centered at the rough relative position.

[0108] Example 40

[0109] The analyzer of example 30, wherein: the first particles are particles from a first aspiration of a blood quality control material; and the second particles are particles from a second aspiration of the blood quality control material.

[0110] Example 41

[0111] The analyzer of example 30, wherein the first particles are particles comprised by a first quality control sample and the second particles are particles comprised by a second quality control sample.

[0112] Example 42- 18 -01337880810419 4919-0466-2881v3

[0113] The analyzer of example 30, wherein the flowcell comprises a flow passage configured to convey the first and second particles, and wherein the target is located on the flowcell and externally of the flow passage.

[0114] Example 43

[0115] A method comprising performing the set of acts the instructions stored on the one or more non-transitory computer readable mediums of the analyzer of any of examples 30-42 are to perform when executed.

[0116] All patents, patent publications, patent applications, journal articles, books, technical references, and the like discussed in the instant disclosure are incorporated herein by reference in their entirety for all purposes.

[0117] Different arrangements of the components depicted in the drawings or described above, as well as components and steps not shown or described are possible. Similarly, some features and sub-combinations are useful and may be employed without reference to other features and sub-combinations. Embodiments of the present disclosure have been described for illustrative and not restrictive purposes, and alternative embodiments will become apparent to readers of this patent. In certain cases, method steps or operations may be performed or executed in differing order, or operations may be added, deleted or modified. It can be appreciated that, in certain aspects of the present disclosure , 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 substitution would not be operative to practice certain embodiments of the present disclosure, such substitution is considered within the scope of the present disclosure. Accordingly, the claims should not be treated as limited to the examples, drawings, embodiments and illustrations provided above, but instead should be understood as having the scope provided when their terms are given their broadest reasonable interpretation as provided by a general -purpose- 19 -01337880810419 4919-0466-2881v3dictionary, except that when a term or phrase is indicated as having a particular meaning under the heading Explicit Definitions, it should be understood as having that meaning when used in the claims.

[0118] Explicit Definitions

[0119] It should be understood that, in the above examples and the claims, a statement that something is “based on” something else should be understood to mean that it is determined at least in part by the thing that it is indicated as being based on. To indicate that something must be completely determined based on something else, it is described as being “based EXCLUSIVELY on” whatever it must be completely determined by.

[0120] It should be understood that, in the above examples and claims, the term “set” should be understood as one or more things which are grouped together.- 20 -01337880810419 4919-0466-2881v3

Claims

CLAIMSWhat is claimed is:

1. A method of operating a biological analyzer comprising: setting a first relative position between a camera and a flowcell by focusing to a target; after setting the first relative position setting a second relative position between the camera and the flowcell based on focusing to first particles passing through the flowcell; and confirming the second relative position based on capturing and analyzing images of second particles passing through the flowcell.

2. The method of claim 1, wherein setting the second relative position comprises moving the camera along an optical axis of the camera.

3. The method of claim 1, wherein: setting the second relative position comprises: setting a rough relative position based on focusing to the first particles passing through the flowcell; and setting a fine relative position based on the rough relative position and on focusing to third particles passing through the flowcell; and the second relative position is the fine relative position.

4. The method of claim 3, wherein: setting the rough relative position comprises moving the camera along an optical axis of the camera at a rough adjustment speed through a rough adjustment range; and setting the fine relative position comprises moving the camera along the optical axis of the camera at a second adjustment speed through a second adjustment range.- 21 -01337880810419 4919-0466-2881v35. The method of claim 4, wherein the second adjustment speed is slower than the rough adjustment speed.

6. The method of claim 4, wherein the second adjustment range is smaller than the rough adjustment range.

7. The method of claim 4, wherein rough adjustment range is greater than 50 micrometers.

8. The method of claim 4, wherein the second adjustment range is under 20 micrometers.

9. The method of claim 4, wherein the rough adjustment range is centered at the first relative position.

10. The method of claim 4, wherein the second adjustment range is centered at the rough relative position.

11. The method of claim 1, wherein: the first particles are particles from a first aspiration of a blood quality control material; and the second particles are particles from a second aspiration of the blood quality control material.

12. The method of claim 1, wherein the first particles are particles comprised by a first quality control sample and the second particles are particles comprised by a second quality control sample.

13. The method of claim 1, wherein the biological analyzer is a urine analysis instrument.- 22 -01337880810419 4919-0466-2881v314. The method of claim 1, wherein the flowcell comprises a flow passage conveying the first and second particles, and wherein the target is located on the flowcell and externally of the flow passage.

15. A biological analysis system comprising: at least one processor; and a non-transitory computer readable medium having stored thereon instruction which, when executed by the at least one processor, cause the system to perform the method of any of claims 1- 14.

16. A biological analysis system comprising: a camera; a flowcell; a target wherein the target has a fixed position relative to the flowcell; at least one processor; and a computer-readable medium storing instructions that, when executed by the at least one processor, perform a method comprising: setting a first relative position between a camera and a flowcell by focusing to the target; after setting the first relative position, setting a second relative position between the camera and the flowcell based on focusing to first particles passing through the flowcell; and confirming the second relative position based on capturing and analyzing images of second particles passing through the flowcell.

17. The system of claim 16, wherein setting the second relative position comprises moving the camera along an optical axis of the camera.- 23 -01337880810419 4919-0466-2881v318. The system of claim 16, wherein: setting the second relative position comprises: setting a rough relative position based on focusing to the first particles passing through the flowcell; and setting a fine relative position based on the rough relative position and on focusing to third particles passing through the flowcell; and the second relative position is the fine relative position.

19. The system of claim 18, wherein: setting the rough relative position comprises moving the camera along an optical axis of the camera at a rough adjustment speed through a rough adjustment range; and setting the fine relative position comprises moving the camera along the optical axis of the camera at a second adjustment speed through a second adjustment range.

20. The system of claim 19, wherein the second adjustment speed is slower than the rough adjustment speed.

21. The system of claim 19, wherein the second adjustment range is smaller than the rough adjustment range.

22. The system of claim 19, wherein rough adjustment range is greater than 50 micrometers.

23. The system of claim 19, wherein the second adjustment range is under 20 micrometers.

24. The system of claim 19, wherein the rough adjustment range is centered at the first relative position.- 24 -01337880810419 4919-0466-2881v325. The system of claim 19, wherein the second adjustment range is centered at the rough relative position.

26. The system of claim 16, wherein: the first particles are particles from a first aspiration of a blood quality control material; and the second particles are particles from a second aspiration of the blood quality control material.

27. The system of claim 16, wherein the first particles are particles comprised by a first quality control sample and the second particles are particles comprised by a second quality control sample.

28. The system of claim 16, wherein the flowcell comprises a flow passage configured to convey the first and second particles, and wherein the target is located on the flowcell and externally of the flow passage.

29. A method comprising performing the set of acts the instructions stored on the one or more non-transitory computer readable mediums of the system of any of claims 16-28 are to perform when executed.

30. An analyzer, comprising: a camera; a flowcell; a target wherein the target has a fixed position relative to the flowcell; at least one processor; and a computer-readable medium storing instructions that, when executed by the at least one processor, perform a method comprising:- 25 -01337880810419 4919-0466-2881v3setting a first relative position between a camera and a flowcell by focusing to the target; after setting the first relative position, setting a second relative position between the camera and the flowcell based on focusing to first particles passing through the flowcell; and confirming the second relative position based on capturing and analyzing images of second particles passing through the flowcell.

31. The analyzer of claim 30, wherein setting the second relative position comprises moving the camera along an optical axis of the camera.

32. The analyzer of claim 30, wherein: setting the second relative position comprises: setting a rough relative position based on focusing to the first particles passing through the flowcell; and setting a fine relative position based on the rough relative position and on focusing to third particles passing through the flowcell; and the second relative position is the fine relative position.

33. The analyzer of claim 32, wherein: setting the rough relative position comprises moving the camera along an optical axis of the camera at a rough adjustment speed through a rough adjustment range; and setting the fine relative position comprises moving the camera along the optical axis of the camera at a second adjustment speed through a second adjustment range.

34. The analyzer of claim 33, wherein the second adjustment speed is slower than the rough adjustment speed.- 26 -01337880810419 4919-0466-2881v335. The analyzer of claim 33, wherein the second adjustment range is smaller than the rough adjustment range.

36. The analyzer of claim 33, wherein rough adjustment range is greater than 50 micrometers.

37. The analyzer of claim 33, wherein the second adjustment range is under 20 micrometers.

38. The analyzer of claim 33, wherein the rough adjustment range is centered at the first relative position.

39. The analyzer of claim 33, wherein the second adjustment range is centered at the rough relative position.

40. The analyzer of claim 30, wherein: the first particles are particles from a first aspiration of a blood quality control material; and the second particles are particles from a second aspiration of the blood quality control material.

41. The analyzer of claim 30, wherein the first particles are particles comprised by a first quality control sample and the second particles are particles comprised by a second quality control sample.

42. The analyzer of claim 30, wherein the flowcell comprises a flow passage configured to convey the first and second particles, and wherein the target is located on the flowcell and externally of the flow passage.- 27 -01337880810419 4919-0466-2881v343. A method comprising performing the set of acts the instructions stored on the one or more non-transitory computer readable mediums of the analyzer of any of claims 30-42 are to perform when executed.- 28 -01337880810419 4919-0466-2881v3

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