Residual monitoring for validating flow cytometer cleaning cycles

Abstract

The present disclosure provides systems and methods for operating a sample processing instrument. The methods include: processing one or more samples by a fluidics system of the sample processing instrument; cleaning the fluidics system; measuring an amount of particles at a detection point of the fluidics system by: selectively supplying a flow from one of a sample tube source and a sample tube line source to the fluidics system, measuring the amount of particles in the flow; comparing the measured amount of particles to a predetermined target value; and using the comparison to determine whether a cleaning requirement is met.

Description

[0001] Cross-references to related applications

[0002] This application is filed as a PCT international application and claims the benefit and priority of U.S. Provisional Application No. 63 / 562,119, filed on March 6, 2024, the disclosure of which is hereby incorporated by reference in its entirety. Background Technology

[0003] Sample processing instruments are used to analyze liquid samples that may include small suspended particles (e.g., biological particles such as extracellular vesicles; non-biological particles such as beads) or cells, and / or to sort particles or cells in said liquid samples. Sample processing instruments typically process multiple samples, and after processing one sample, the instrument needs to be cleaned to avoid inaccurate results for the next sample.

[0004] To prevent contamination between samples due to carryover of particles that may remain in the system after sample processing, many sample handling instruments are equipped with cleaning cycles to remove residual particles and ensure the cleanliness of the sample tubing. However, with conventional sample handling instruments, it is not easy for users to accurately monitor and understand the cleaning results. This is disadvantageous for processing samples, especially those containing small particles (e.g., nanoparticles) that are difficult to clean. Summary of the Invention

[0005] The examples presented herein relate to a method for operating a sample processing instrument. The method includes: processing one or more samples via a flow control system of the sample processing instrument; cleaning the flow control system; measuring the amount of particles at a detection point of the flow control system by selectively supplying a flow from either a sample tubing source or a sample line source to the flow control system, measuring the amount of particles in the flow; comparing the measured amount of particles with a predetermined target value; and using the comparison to determine whether a cleaning requirement is met.

[0006] In other examples presented herein, the flow is supplied from the sample tube source, and it is determined that the cleaning requirement is not met. The method further includes: supplying a second flow from the sample tube source; measuring a second particle amount in the second flow; comparing the measured second particle amount with the predetermined target value; and using the comparison to determine whether the cleaning requirement is met. In still other examples presented herein, the source of the flow is determined based on user input. In yet another example presented herein, the predetermined target value differs based on whether the flow is supplied from the sample tube source or from the sample tube source. In yet another example presented herein, the predetermined target value is higher for a flow supplied from the sample tube source and lower for a flow supplied from the sample tube source.

[0007] In other examples presented herein, the sample tubing source comprises a sheath fluid. In yet another example presented herein, the sheath fluid is a 5-nanometer (nm) sheath fluid. In other examples presented herein, the sample tubing source is a sample buffer. In still another example presented herein, the predetermined target value is 100 events.

[0008] Other examples presented herein relate to a system for operating a sample processing instrument. The system includes: a flow control system comprising a flow chamber, a sample tube source, a sample tubing source, and a flow control path leading to the flow chamber; a source selection valve communicating with the flow control path and selectively communicating with each of the sample tube source and the sample tubing source; and a controller including a processor and a memory. The memory includes instructions that, when executed by the processor, cause the processor to: generate a display including a source selection operation; receive input from the source selection operation; and use the input to operate the source selection valve to provide flow from one of the sample tube source and the sample tubing source to the flow chamber.

[0009] In other examples presented herein, the source selection operation is associated with determining whether a cleaning requirement is met based on the particle quantity measured at a detection point in the flow control system. In yet another example presented herein, the cleaning requirement includes a predetermined threshold particle quantity. In yet another example presented herein, the predetermined threshold particle quantity is 100 events.

[0010] In other examples presented herein, the sample tubing source is sheath fluid. In yet another example presented herein, the sheath fluid is 5 nm sheath fluid. In still other examples presented herein, the sample processing instrument is a flow cytometer.

[0011] Other examples presented herein relate to a system for operating a sample processing instrument. The system includes at least one processor, a memory communicating with the processor, and instructions that, when executed by the at least one processor, cause the processor to: determine whether the flow control system of the sample processing instrument meets cleanliness requirements based on the amount of particles present in the measurement stream; and generate a graphical user interface including a source selection operation, wherein the source selection operation accepts input from a user selecting either a sample tube source or a sample tubing source as the source of the measurement stream.

[0012] In other examples presented herein, the sample source is a sample buffer, and the sample tubing source is a sheath fluid. In another example presented herein, the sheath fluid is a 5 nm sheath fluid. In yet another example presented herein, the cleaning requirement includes a threshold particle amount, and the first threshold particle amount of the sample tubing source is an amount smaller than the second threshold particle amount of the sample source.

[0013] Several other aspects of the invention will also be set forth in the following description. These aspects may relate to individual features as well as combinations of features. It should be understood that the foregoing general description and the following detailed description are merely exemplary and explanatory, and do not limit the broad inventive concept upon which the embodiments disclosed herein are based. Attached Figure Description

[0014] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate several aspects of this disclosure. A brief description of the drawings is as follows:

[0015] Figure 1 This is a functional block diagram of an example sample processing instrument.

[0016] Figure 2 This is a schematic diagram of the flow control system of a sample processing instrument according to an embodiment of this application.

[0017] Figure 3A This is the first in a series of figures illustrating an example use case of using sheath fluid as a monitoring fluid.

[0018] Figure 3B yes Figure 3A The second figure in a series of illustrations shows an example use case of using sheath fluid as a monitoring fluid.

[0019] Figure 3C yes Figure 3A The third figure in a series of diagrams shows an example use case of using sheath fluid as a monitoring fluid.

[0020] Figure 3D yes Figure 3A The fourth figure in a series of illustrations shows an example use case of using sheath fluid as a monitoring fluid.

[0021] Figure 4 This is a flowchart of an example method for operating a sample processing instrument.

[0022] Figure 5 This is a flowchart of an example method for measuring particle quantity in a flow control system.

[0023] Figure 6 This is a sample user interface that enables users to define and perform cleaning and monitoring operations for sample handling systems.

[0024] Figure 7 yes Figure 6 Example graphical user interface for parameter setting elements.

[0025] Figure 8 An example of a control unit that can be used to implement the aspects described herein is illustrated schematically. Detailed Implementation

[0026] The present application is described in detail below with reference to the accompanying drawings and exemplary embodiments. In several drawings, similar reference numerals indicate similar parts and components. The following detailed description of the present application is for explanation only and is in no way intended to limit the present application or its application or use. The embodiments described in this specification are not exhaustive, but are merely some of the many possible embodiments. Exemplary embodiments may be implemented in many different forms and should not be construed as limiting the scope of the present application. In some exemplary embodiments, well-known methods, well-known apparatus structures, and well-known techniques may not be described in detail.

[0027] Before explaining at least one embodiment of this application in detail, it should be understood that this application is not limited in its application to the details of the construction and arrangement of the components set forth in the following description or shown in the accompanying drawings. The invention is applicable to other embodiments that can be practiced or implemented in various ways, as well as combinations of the disclosed embodiments. Furthermore, it should be understood that the wording and terminology used herein are for illustrative purposes and should not be considered restrictive.

[0028] The sample processing instrument according to this disclosure will be described using a flow cytometer as an example. A flow cytometer is used to detect particles in a sample to determine one or more properties of the particles. However, it should be understood that the sample processing instrument according to this disclosure is not limited to a flow cytometer and can be any other suitable instrument for processing biological or non-biological samples. In some embodiments, the sample processing instrument may be a cell or particle sorter.

[0029] The sample processing instrument according to this disclosure is suitable for automatically performing cleaning processes between different sample processing steps, for automatically performing cleaning processes using different cleaning agents, and for automatically performing a monitoring process to measure the cleaning results after the cleaning process. The system can further automatically determine the next action based on the measurement results. For example, various monitoring agents are discussed herein, which have different sensitivities to measuring particles at detection points in the flow control system. Additionally, the system can provide a user interface to allow easy operation and intuitive observation of the monitoring and cleaning processes. For example, a user interface is provided, allowing the user to select the monitoring agent to be used to measure residual particles at detection points in the flow control system. As discussed herein, residual particles refer to sample particles and other debris that may remain in the flow control system after sample processing and contaminate future sample analyses.

[0030] The following will refer to Figure 1 The main functional components of sample processing instrument 1 are described. Samples processed (e.g., analyzed or sorted) by sample processing instrument 1 may include biological particles (such as exosomes or extracellular vesicles) or non-biological particles (such as beads). The disclosed system is optimized for the detection and measurement of nanoscale particles (e.g., nanoparticles, nanobeads, exosomes), but the disclosed system can also be used for larger particles.

[0031] Figure 1 This is a functional block diagram of sample processing instrument 1. (For example...) Figure 1 As shown, the sample processing instrument 1 includes a flow control component 10, a flow chamber 20, a sample processing unit 30, and a control unit 40.

[0032] The flow control assembly 10 is configured to supply various fluids to and discharge fluids from the flow chamber 20, which includes a detection point. Fluids described herein may include samples to be analyzed, sorted, or otherwise processed, sheath fluids, cleaning agents, waste liquids, etc. The flow control assembly 10 may include various pumps, valves, pressure regulators, sensors, etc., for delivering or discharging fluids. Various fluids, particularly samples and sheath fluids, are delivered to the flow chamber 20.

[0033] Figure 2 This is a schematic diagram of the flow chamber of a sample processing apparatus according to an embodiment of this application. (Reference) Figure 2The flow chamber 20 includes two opposing sheath fluid ports 21 and 22, through which sheath fluid is delivered into chamber 25 of the flow chamber 20. The flow chamber 20 further includes a sample needle 23 disposed therein, through which a sample is delivered into chamber 25. In chamber 25, the sample is encapsulated in sheath fluid and then flows through a cuvette 26 for processing. The cuvette 26 forms a processing area for the sample and may include a detection point. For example, an optical detection device focuses a light beam into the processing area (e.g., the detection point) of the cuvette 26. As particles in the sample pass through the processing area of ​​the cuvette 26, the characteristics of the particles are determined by measuring the light scattered or emitted from the particles.

[0034] The sample processing unit 30 processes a sample encapsulated in a sheath fluid flowing through the cuvette 26. For example, the sample processing unit 30 can measure the characteristics of particles / cells in the sample and quantify particles / cells with specific characteristics, and / or the sample processing unit 30 can sort the particles / cells based on their characteristics. Depending on the purpose of sample processing, the sample processing unit 30 may include various optical, electrical, and / or mechanical devices.

[0035] The control unit 40 controls the operation of the entire sample processing instrument 1. The various functions, actions, or steps of the various systems, devices, components, or methods of the sample processing instrument according to this application are controlled by the control unit 40. The control unit 40 will be described in detail below.

[0036] The following will refer to Figure 2 Examples of flow control components according to embodiments of this application are described. Figure 2 A portion of system 100 is shown, which includes flow control devices for controlling the flow of different fluids. As described above, the flow control system 100 is used to supply various fluids to and discharge fluids from the flow chamber 20. To this end, the flow control system 100 includes fluid lines connecting various fluid sources to the flow chamber 20.

[0037] These fluid sources may include a sample source 101, a sheath fluid source (not shown), a waste liquid reservoir, and other solution sources. The sample source 101 is used to supply samples. Typically, the sample source 101 includes multiple sample containers containing different samples, such as well plates, test tubes, etc. The sheath fluid is stored in the sheath fluid source. The sheath fluid is a matrix liquid that facilitates the proper detection of the sample flow and can be used to encapsulate the sample flow and keep it centered in the nozzle to ensure detection accuracy, while preventing particles in the sample flow from approaching the nozzle wall and clogging the nozzle. Additionally, the sheath fluid can be used as a cleaning agent for cleaning sample processing instruments (particularly flow chambers and fluid lines) and / or as a monitoring solution for monitoring the presence of residues in the system. Other solution sources include a fluid container 103 storing other cleaning agents besides the sheath fluid (e.g., water or another special cleaning solution), and a separate container (not shown) storing a monitoring solution (e.g., water or buffer solution) for measuring the cleaning results of the sample processing instruments. The waste liquid reservoir is used to collect waste liquid after sample processing and cleaning of the sample processing instruments.

[0038] refer to Figure 2 The fluid lines include sample lines 111, 112, and 113 connecting sample source 101 to sample needle 23; sheath fluid line 117 connecting sheath fluid source (not shown) to sheath fluid ports 21 and 22 of flow chamber 20; waste fluid line 116 connecting flow chamber 20 to waste fluid reservoir (not shown); sheath fluid cleaning lines 153 and 154 for delivering sheath fluid for cleaning; and cleaning agent lines 142 and 144 for delivering cleaning agent for cleaning.

[0039] Various pumps can be installed in fluid pipelines to pump various fluids. Figure 2 In the example shown, there is a sample pump 121 for pumping samples, a sheath fluid pump 125 for pumping sheath fluid for cleaning, and a cleaning pump 123 for pumping cleaning agents. Figure 2 In the examples, sample pump 121, sheath pump 125, and cleaning pump 123 are all piston pumps. However, it should be understood that the system disclosed in this application is not limited to the specific examples shown in the drawings, as long as it can achieve the functions described herein. For example, the type of pump can be changed. In embodiments, sheath pump 125 and cleaning pump 123 can be other types of pumps, such as peristaltic pumps. Similarly, sample pump 121 can also be any other suitable type of pump, such as a peristaltic pump. In some embodiments, the number of pumps can be changed. In some instances, the sheath pump is omitted.

[0040] Various switching devices can be installed in fluid pipelines, such as those for switching the flow direction of fluid or controlling the on / off state of fluid. Switching devices can include various types of valves. For example... Figure 2As shown, the switching device includes three-way valves 131 and 132 and switching valves 141, 151 and 152.

[0041] The three-way valve 132 is configured to selectively connect sample lines 111 to 113 to different pumps (e.g., sample pump 121 or cleaning pump 123) to aspirate or pump different fluids (e.g., sample or cleaning agent) to flow chamber 20 or sample source 101. Figure 2 In this example, the three-way valve 132 includes a first port 1321 connected to the sample line 113, a second port 1322 connected to the sample pump 121, and a third port 1323 connected to the cleaning pump 123. When the first port 1321 is switched to be connected to the second port 1322, the sample pump 121 is allowed to aspirate a sample from the sample source 101 when the sample line 113 is connected to the sample line 111, or the fluid (e.g., sample or sheath fluid) is allowed to be pumped into the flow chamber 20 when the sample line 113 is connected to the sample line 112. That is, the system can allow the sample pump 121 to aspirate a sample from the sample source 101 by switching valve 132 to connect the sample pump 121 to the sample line 113 and by switching valve 131 to connect the sample line 113 to the sample line 111 (alternatively, the sample or sheath fluid can be sent into the flow chamber 20 by switching valve 131 to connect the sample line 113 to the sample line 112). The system allows the cleaning pump 123 to draw cleaning agent from the fluid container 103 and pump it into the flow chamber 20 via the switching valve 132 to connect the cleaning line 144 to the sample line 113 and via the switching valve 131 to connect the sample line 113 to the sample line 112. (Alternatively, the cleaning agent can be sent to the sample source 101 via the switching valve 131 to connect the sample line 113 to the sample line 111.)

[0042] The three-way valve 131 is configured to selectively connect a pump (e.g., sample pump 121 or cleaning pump 123) to sample source 101 or flow chamber 20 to selectively aspirate or pump fluid (e.g., sample or cleaning agent) to flow chamber 20 or sample source 101. Figure 2In this example, a three-way valve 131 is positioned between sample lines 111, 112, and 113 for selectively communicating sample line 113 with either sample line 111 or sample line 112. The three-way valve 131 has a first port 1311 connected to sample pump 121 or cleaning pump 123 via a three-way valve 132, a second port 1312 connected to sample needle 23, and a third port 1313 connected to sample source 101. The system may allow fluid (e.g., sample pumped by sample pump 121 or cleaning agent pumped by cleaning pump 123) in sample line 113 to be delivered to flow chamber 20 by switching valve 131 to connect sample line 113 to sample line 112, or alternatively, allow fluid to be aspirated from / pumped into sample source 101 by switching valve 131 to connect sample line 113 to sample line 111.

[0043] Three-way valves 131 and 132 can selectively pump samples to flow chamber 20 for example to analyze samples, or pump cleaning agents to sample source 101 or flow chamber 20 to clean sample lines 111 to 113 or flow chamber 20.

[0044] The cleaning pump 123 is connected to the third port 1323 of the three-way valve 132 via the cleaning agent line 144, and is also connected to the fluid container 103 via the cleaning agent line 142. A switch valve 141 can be installed in the cleaning agent line 142 to control its on / off state. When cleaning agent is being pumped, the switch valve 141 is closed to allow the cleaning agent line 142 to flow. When cleaning agent is not being pumped, the switch valve 141 is open to disconnect the cleaning agent line 142.

[0045] A sheath fluid pump 125 is arranged between a sheath fluid line 117 connected to a sheath fluid source and a sample pump 121 to deliver sheath fluid to the sample source 101 or flow chamber 20 via the sample pump 121 for cleaning the sample lines 111-113 or flow chamber 20 with sheath fluid. The sheath fluid pump 125 is connected to the sheath fluid line 117 (or the sheath fluid source) via a sheath fluid cleaning line 153 and to the sample pump 121 via a sheath fluid cleaning line 154. A switch valve 151 can be provided in the sheath fluid cleaning line 153 to control its on / off state. When sheath fluid is aspirated for cleaning, the switch valve 151 is in the closed (i.e., open) state to allow the sheath fluid cleaning line 153 to be connected. When sheath fluid aspiration is not required, the switch valve 151 is in the open (i.e., closed) state to interrupt the connection of the sheath fluid cleaning line 153. Furthermore, a switch valve 152 can be installed in the sheath fluid cleaning line 154 to control the on / off state of the sheath fluid cleaning line 154. When sheath fluid is being pumped, the switch valve 152 is in the closed (i.e., open) state to allow the sheath fluid cleaning line 154 to be connected. When sheath fluid pumping is not required, the switch valve 152 is in the open (i.e., closed) state to interrupt the connection of the sheath fluid cleaning line 154.

[0046] In this embodiment, this flow path is also used to supply sheath fluid to be used as a monitoring fluid.

[0047] It should be understood that the system according to this disclosure is not limited to... Figure 2 The specific examples shown are not definitive and can be modified according to actual requirements. For example, the sheath fluid pump and cleaning pump can be peristaltic pumps, and therefore the switching valves in the sheath fluid cleaning line and cleaning agent line can be omitted. In another example, the sheath fluid pump is omitted, and instead, only the switching valve is provided in the sheath fluid cleaning line between the sheath fluid line (or sheath fluid source) and the sample pump. It should be understood that the system according to this application is not limited to having the components described above. For example, the system may also have a filter (e.g., such as...). Figure 2 The illustrated components include a filter 119 for filtering sheath fluid, a sensor for sensing temperature or pressure, and a regulator for adjusting temperature or pressure. In some preferred embodiments, filter 119 is a 5-nanometer (nm) filter.

[0048] The following will refer to Figure 2 An example process for delivering a sample via flow control system 100 during sample processing is described. When a sample is to be processed, three-way valve 132 is switched so that sample pump 121 is connected to sample line 113, and three-way valve 131 is switched so that sample line 113 is connected to sample line 111, thereby drawing the sample from sample source 101 into sample line 113 via sample pump 121 (e.g., the piston of sample pump 121 moves downward).

[0049] Then, the three-way valve 131 is switched so that the sample line 113 is connected to the sample line 112, and the sample in the sample line 113 is pumped to the flow chamber 20 for processing (e.g., detection or sorting) by the sample pump 121 (e.g., the piston of the sample pump 121 moves upward).

[0050] During sample processing, sample pump 121 is always connected to sample line 113, and three-way valve 131 repeatedly switches between second port 1312 and third port 1313 to repeatedly perform the process of aspirating and pumping the sample until sample processing is complete.

[0051] After sample processing, it is necessary for the flow control system to clean the sampling flow path to prevent contamination of future samples and maintain proper system operation.

[0052] The following text will continue to refer to Figure 2 The process of the flow control system 100 cleaning sample lines 111 to 113 and flow chamber 20 with a cleaning agent is described. When the sample processing instrument is to be cleaned with a cleaning agent, the switch valve 141 is closed to connect the cleaning agent line 142, thereby allowing the cleaning pump 123 to draw cleaning agent from the fluid container 103.

[0053] When the switch valve 141 is switched to the off state and the three-way valve 132 is switched, the cleaning agent line 144 is connected to the sample line 113 to pump the cleaning agent into the sample line 113. At this time, the three-way valve 131 can be in a state where the sample line 113 is connected to either the sample line 112 or the sample line 111.

[0054] With sample line 113 connected to sample line 111, cleaning agent is pumped through sample line 111 to clean sample lines 113 and 111. Next, the switch valve 141 is repeatedly closed or opened to aspirate or pump cleaning agent until sample line 111 is cleaned.

[0055] With sample line 113 connected to sample line 112, cleaning agent is pumped through sample line 112 and flow chamber 20, thereby cleaning sample lines 113 and 112 and flow chamber 20. Next, the switch valve 141 is repeatedly closed or opened to aspirate or pump cleaning agent until the flow control system, including sample line 112 and flow chamber 20, is cleaned.

[0056] The following text will continue to refer to Figure 2 The process of using sheath fluid to clean sample lines 111 to 113 and flow chamber 20 in flow control system 100 is described. When cleaning the sample processing instrument with sheath fluid, switching valve 151 is closed to connect sheath fluid cleaning line 153, thereby allowing sheath fluid pump 125 to draw sheath fluid from sheath fluid source (not shown).

[0057] Then, switch valve 151 is placed in the open state and switch valve 152 is placed in the closed state, and three-way valve 132 is switched so that the first port 1321 is connected to the second port 1322, so that the sheath fluid is pumped into sample line 113 by sample pump 121. Three-way valve 131 can be in a state where sample line 113 is connected to either sample line 112 or sample line 111.

[0058] With sample line 113 connected to sample line 111, sheath fluid is pumped through sample line 111 to clean sample lines 113 and 111. Next, switching valves 151 and 152 are alternately closed or opened to aspirate or pump sheath fluid until the sample lines are cleaned.

[0059] With sample line 113 connected to sample line 12, sheath fluid is pumped through sample line 112 and flow chamber 20, thereby cleaning sample lines 113 and 112 and flow chamber 20. Next, switching valves 151 and 152 are alternately closed or opened to aspirate or pump sheath fluid until the sample lines and flow chamber 20 are cleaned.

[0060] In addition to the sample processing and cleaning processes described above, the system can also be used to monitor the cleanliness of the sample processing instrument. After system cleaning, monitoring operations are performed to ensure that debris and contaminants are adequately removed from the system. In embodiments, multiple monitoring fluids can be used in the sample processing system. One or more monitoring fluids can be used to perform monitoring operations. In examples, one or more monitoring fluids can be used sequentially and automatically, or they can be used in response to user selection.

[0061] In some embodiments, between analyses of two different samples, a monitoring solution (e.g., water, buffer solution, sheath fluid) can be used instead of the sample to fill sample source 101. For example, the first sample in sample source 101 can be analyzed by system 100, which can be cleaned with a cleaning agent from fluid container 103, and then sample source 101 can be switched to a different sample source 101 filled with the monitoring solution to monitor for the presence of residue in flow chamber 20. In some cases, the monitoring solution and cleaning agent can be the same fluid, such as water.

[0062] In some embodiments, the cleaning agent may be a monitoring solution, in which case the fluid container 103 may serve as a source of both the monitoring solution and the cleaning agent. In this case, the process of feeding the monitoring solution through the flow chamber 20 and other components of the flow control system during the monitoring process may be similar to the process of feeding the cleaning agent through the flow chamber 20 and the flow control system during the cleaning process.

[0063] In some embodiments, a separate fluid container (i.e., a container other than sample source 101 and fluid container 103) may be provided to contain the monitoring solution. In this case, the same pump used for other fluids or an additional pump may be used to pump the monitoring solution through the detection point of the flow control system, such as flow chamber 20. The fluid line for the monitoring solution may be integrated into other fluid lines (such as a sheath fluid cleaning line), or it may be a separate fluid line from the fluid container containing the monitoring solution to flow chamber 20. Similarly, the sheath fluid cleaning line may also be formed as a separate fluid line from the sheath fluid source to flow chamber 20 and the sample line, that is, the sheath fluid cleaning line does not pass through sample pump 121.

[0064] In some preferred embodiments, the sheath fluid is used as a monitoring solution. In this case, the sheath fluid pump 125 or another pump can be used to pump the monitoring solution through the flow chamber 20. The sheath fluid line 117 can also, or alternatively, be used to supply the sheath fluid to the flow chamber for use as a monitoring solution.

[0065] Figure 3A -D is a series of diagrams illustrating example use cases of using sheath fluid as a monitoring fluid. Figure 3A , 3C Figures 202, 206, and 208 (and 3D respectively) depict the use of sample tubes (such as...) Figure 2 The residual readings were obtained from the flow cytometer using the monitoring fluid of sample source 101. In each case, some significant residues were detected. However, Figure 3B Figure 204 depicts the residual readings obtained using a monitoring fluid (such as a sheath fluid, which can be a 5 nm sheath fluid) from the sample tubing. In this case, significantly less residue was measured. The difference in residual measurements can be attributed to the different monitoring fluids chosen when using common equipment and settings to collect each column of readings.

[0066] As the sensitivity of sample processing instruments increases, buffer solutions and other sample tube fluids may carry excessively high numbers of particles and other debris, making it impossible to provide an accurate measurement of instrument cleanliness. In other words, debris in the buffer solution itself may provide deceptively high residual readings. A more accurate measurement of residual material within the instrument can be achieved by using a cleaner monitoring fluid, such as a sheath fluid (e.g., a 5 nm sheath fluid). In this way, while the user's ability to control or verify the cleanliness of the sample buffer solution and other fluids that can be introduced via sample tubes is limited, the cleanliness of the instrument can be verified.

[0067] As stated above, the structure of the disclosed system and its flow control components are not limited to the specific examples described and shown above, but can be modified as long as they enable an automated cleaning / monitoring process or an automated cleaning process using different cleaning agents. Furthermore, since the structure of the disclosed system can be modified, the operating method of the disclosed system can be modified accordingly.

[0068] Figure 4 This is a flowchart of an example method 300 for operating a sample processing instrument. Method 300 can be performed by a sample processing instrument (such as...) Figure 1 The sample processing instrument 1, which may be a flow cytometer, is used for execution. In an embodiment, method 300 may be performed by a control unit (such as...). Figure 1 The automated process is executed by the control unit 40.

[0069] At operation 302, one or more samples are processed via the flow control system of a sample processing instrument. For example, a first sample, which may contain multiple particles, is fed into a flow chamber and processed via the flow path of the flow control system (as discussed above). The particles in the first sample are detected or sorted by the instrument. The particles are, for example, biological nanoparticles.

[0070] After the first sample has been processed, it may be necessary to process another sample. To obtain accurate results, it may be necessary to clean the flow chamber and one or more lines of the system between consecutive samples to prevent particles from the first sample from affecting the results of processing the second sample. These residual particles (referred to herein as residues) are often difficult to remove, especially when the particle size is small. Therefore, to ensure accurate processing of the second sample, the sample processing instrument must be thoroughly cleaned. In this embodiment, the flow chamber and sample flow path of the flow control system may require particular attention.

[0071] At operation 304, the clean flow control system is cleaned. Suitable cleaning agents (e.g., sheath fluid, water, and / or any other suitable cleaning solution), cleaning parameters (e.g., duration of a single cleaning cycle, number of cleaning cycles, maximum number of cleaning cycles, etc.), monitoring solutions (e.g., water or sheath fluid), and / or monitoring parameters can be selected or set. Monitoring parameters may include parameters associated with the monitored solution (e.g., delivery time or volume, etc.) and population or monitoring parameters associated with the monitored particles (e.g., monitoring criteria indicating compliance with cleaning requirements). In embodiments, one or more of the cleaning agent, cleaning parameters, monitoring solutions, and monitoring parameters may be set based on known or identifiable characteristics of the particles in the first sample. Monitoring criteria may be embodied in various forms, such as target residue counts over a predetermined time, target residue concentration / concentration percentage, target residue rate (numbers / second), etc. Example settings for cleaning parameters, as well as monitoring parameters and criteria, can be found in PCT Publication 2023 / 065796, the entire contents of which are incorporated herein by reference.

[0072] The sample processing instrument cleans itself using selected cleaning agents according to pre-defined cleaning parameters. During the cleaning cycle, one, two, or more cleaning agents can be selected. Correspondingly, cleaning parameters can be set for each cleaning agent. These parameters can be determined based on experimental data, historical data, or other experimental data.

[0073] At operation 306, the particle quantity at the detection point of the flow control system is measured. The flow control assembly pumps the monitoring solution through the flow control system. The sample processing instrument then analyzes the detection point, which is typically located within a flow chamber containing the monitoring solution during the monitoring period.

[0074] Figure 5 This is a flowchart of an example method 400 for measuring particle quantity in a flow control system. Method 400 can be performed by a sample processing instrument (such as...) Figure 1 The sample processing instrument 1, which may be a flow cytometer, is used for execution. In an embodiment, method 400 may be performed by a control unit (such as...). Figure 1 The automated process is executed by the control unit 40.

[0075] At operation 402, flow is selectively supplied to the flow control system from either a sample tube source or a sample tubing source. As discussed herein, the sample tubing source refers to a sheath fluid, such as a 5 nm sheath fluid, and the sample tube source is a sample buffer. In this embodiment, the source of the flow is determined based on user input.

[0076] At operation 404, the particle quantity at the detection point of the flow control system is measured. Measurements related to residues or the monitoring solution are obtained during the monitoring period, such as the amount of residue and / or the flow rate of the monitoring solution. The flow rate of the monitoring solution can be measured by one or more sensors. For example, for a sample processing instrument as a flow cytometer, the measurement can be performed by measuring the light scattered from the detection point in response to one or more laser beams directed at the detection point, which may be located within the flow chamber of the flow control system. In this example, residues can be counted, for example, by an optical detection system based on the detected light scattered or emitted from the particles. The lower the amount of residue (e.g., a count or approximate count of the number of residual particles remaining in the flow chamber during the monitoring period) within the same monitoring time, the better the cleaning result. Of course, a longer monitoring time results in a larger amount of residue being detected. If the measured value is insufficient to indicate the level of cleanliness, a value that accurately indicates the level of cleanliness can be calculated. For example, the value may be a residual rate (number / second) (the number of residual particles detected during monitoring divided by the monitoring time), a residual concentration (number / µL) (the number of residual particles detected during monitoring divided by the volume of the monitoring solution), or a residual concentration percentage (the ratio of the residual concentration to the concentration of the first particle in the first sample).

[0077] At operation 406, the measured particle quantity is compared with a predetermined target value. In some embodiments, the target value may be based on a first sample, a second sample, or the type of processing being performed. For example, different samples or different sample processing / analysis may have different target values.

[0078] In this embodiment, the predetermined target value differs based on whether the flow is supplied from a sample tube source or a sample line source. For example, the predetermined target value is higher for a flow supplied from a sample tube source and lower for a flow supplied from a sample line source. In this embodiment, the predetermined target value is 100 events.

[0079] At operation 408, the results of the comparison are used to determine whether the cleaning requirements have been met. If the measured or calculated value (actual value) is less than or equal to the target value, this indicates that the cleaning requirements have been met, and the monitoring process can end at operation 420. In this case, ending the monitoring process includes indicating that the machine's cleanliness is satisfactory and suitable for further sample runs.

[0080] If the measured or calculated value (actual value) is greater than the target value, this indicates that the cleaning requirements have not yet been met, and the process proceeds to operation 410. If it is determined at operation 408 that the cleaning requirements have not been met, further determination can be made at operation 410, namely, whether a sample tubing is used to supply the monitoring fluid, for example, whether the monitoring fluid is a sheath fluid. If it is determined that a sample tubing source is used to supply the flow, additional verification measurements can be performed.

[0081] If the sample tubing is determined to be used as the monitoring fluid at operation 410, the monitoring process ends with unsatisfactory results at operation 422. Unsatisfactory results may indicate that further cleaning or other maintenance is required before any further sample runs.

[0082] If the sample tube is determined to be used as the monitoring fluid at operation 410, additional monitoring measurements can be performed using the sample tube at operation 412. Since sheath fluid, particularly 5 nm sheath fluid, has a known level of cleanliness compared to buffer solutions supplied from the sample tube, the sample tube can be used to assess the presence of the measured residue in the instrument or the monitoring fluid. At operation 412, an additional sheath fluid stream is supplied from the sample tube to the detection point of the flow control system. At operation 414, a second measurement is performed on the particle count in the additional sheath fluid stream. At operation 416, the second measurement is compared to a predetermined target value.

[0083] At operation 418, the comparison results are used to determine whether the cleaning requirements have been met. If the measured value is less than the threshold, the monitoring operation can end at operation 420 with a satisfactory level of system cleanliness. This indicates that the system is ready to process additional samples. In an embodiment, the user can be notified that the cleaning requirements have been met. In some cases, additional samples can then be automatically fed through the flow control system and processed at the detection point.

[0084] If the measured value is equal to or greater than the threshold, the monitoring operation at operation 422 may end with an unsatisfactory state of system cleanliness. This may indicate that further cleaning or other maintenance of the system is required before running another sample. In embodiments, it may be further determined whether a maximum cleaning limit has been reached, such as a set maximum cleaning duration limit (e.g., a maximum limit on the total amount of time spent in one or more cleaning cycles) or a maximum cleaning cycle limit (e.g., a maximum limit imposed on the number of cleaning cycles performed). In some embodiments, the maximum cleaning limit may be set by the user. In some embodiments, the maximum cleaning limit may be based on the first sample, an additional or next sample, or the type of processing in progress. For example, different samples or different sample processing / analysis may have different maximum cleaning limits. If the maximum cleaning limit is not reached, the system may continue the cleaning process until the cleaning requirements are met or the maximum cleaning limit is reached. If the maximum cleaning limit is reached, the cleaning process stops. A message or warning may be issued to the user so that the user can take appropriate actions, such as troubleshooting.

[0085] To facilitate user operation and information access, the method of this application can be implemented through a user interface. Figure 6This is a sample user interface 500 that enables users to define and perform cleaning and monitoring operations on a sample handling system.

[0086] refer to Figure 6 The user interface 500 may include a menu 510, a parameter setting element 520, a monitoring element 540, a sample setting element 530, a historical data viewing element 550, and a control element 560. The menu 510 may include one or more follow-up activity elements that allow the user to interact with the sample processing instrument by configuring sample processing and cleaning of the sample processing instrument, or monitoring such processing or cleaning. The parameter setting element 520, monitoring element 540, sample setting element 530, and historical data viewing element 550 may be displayed on the user interface in response to the selection or operation of the corresponding follow-up activity element of the menu 510, allowing the user to input information or display information to the user.

[0087] Parameter setting element 520 is used to receive user input related to cleaning, monitoring, and residue (which will be referred to below). Figure 7 (Detailed Description). Monitoring element 540 is configured to display information to the user related to cleaning or monitoring status, monitoring data, monitoring results, monitoring standards, etc. The applicable sample setting element 530 includes the sample to be monitored, and the user can select samples for which settings are applied at parameter setting element 520, allowing the sample processing instrument to automatically and continuously process multiple samples. The historical data viewing element 550 allows the user to retrieve or view historical monitoring data upon request. The control element 560 allows the user to control various elements displayed on the user interface, or to display the content of various elements.

[0088] like Figure 6 As shown, menu 510, parameter setting element 520, sample setting element 530, monitoring element 540, historical data viewing element 550, and control element 560 can be displayed simultaneously on one screen, for example, within their respective boxes. It should be understood that the user interface according to this application is not limited to... Figure 6 The specific examples shown are not definitive and can be changed as needed. In some embodiments, any subset of these interface elements may be displayed on the user interface simultaneously. For example, in some cases, only menu 510 and parameter setting element 520 may be displayed on user interface 500 simultaneously. In some embodiments, the interface elements displayed on the user interface may be selected based on user input. For example, the user may select the applicable setting sample element 530, and in response, the user interface may display an enlarged version of the applicable setting sample element 530, without displaying other interface elements (or only displaying a subset of other interface elements). For example, the history data viewing element 550 is optional. Furthermore, the layout of various elements on the user interface can be changed. The content and display format of each element can also be changed as needed.

[0089] Figure 7 yes Figure 6 Example graphical user interface 600 for parameter setting element 520. (e.g., ...) Figure 7 As shown, the parameter setting elements can display multiple menu items to establish parameters for cleaning and monitoring the instrument. The monitoring fluid selection source 602 provides options for selecting a sample tube or sample line as the source. The acceptance criterion selection 604 allows setting a threshold for comparison with the measured particle number (or ratio). The cleaning cycle setting 606 allows, for example, establishing the number of times the cleaning cycle can be repeated in an attempt to obtain satisfactory results.

[0090] Figure 8 The control unit 40 of the sample processing instrument 1, which can be used to implement the aspects described herein, is illustrated schematically. Figure 1 An example of a control unit 40. For example... Figure 8 As shown, the control unit 40 includes one or more processing devices 802, a memory storage device 804, and a system bus 806 coupling the memory storage device 804 to the one or more processing devices 802. The one or more processing devices 802 may include a central processing unit (CPU). In some cases, the one or more processing devices 802 are part of a processing circuitry system having a memory for storing instructions that, when executed by the processing circuitry system, cause the processing circuitry system to perform the various aspects, features, and functions described herein.

[0091] like Figure 8 As shown, the memory storage device 804 may include random access memory (“RAM”) 808 and read-only memory (“ROM”) 810. Basic input and output logic, such as basic routines that help transfer information between elements within the control unit 40 during startup, may be stored in the ROM 810.

[0092] The control unit 40 may further include a mass storage device 812, which may include an operating system 814 and store software instructions and data 816. The mass storage device 812 is connected to the processing device 802 via a system bus 806. The mass storage device 812 and the associated computer-readable data storage medium provide non-volatile, non-transitory storage for the control unit 40.

[0093] Although the description of computer-readable data storage medium contained herein refers to mass storage device 812, those skilled in the art will understand that a computer-readable data storage medium can be any available non-transitory physical device or article of manufacture from which the control unit 40 can read data and / or instructions. A computer-readable storage medium can consist entirely of non-transitory media. Mass storage device 812 is an example of a computer-readable storage device.

[0094] Computer-readable data storage media include volatile and non-volatile, removable and non-removable media implemented in any method or technology for storing information such as computer-readable software instructions, data structures, program modules or other data. Examples of computer-readable data storage media include, but are not limited to, RAM, ROM, EPROM, EEPROM, flash memory or other solid-state memory technologies, or any other medium that can be used to store information and can be accessed by a device.

[0095] The control unit 40 can operate in a networked environment using logical connections to other devices via network 820. The control unit 40 is connected to network 820 via a network interface unit 818 connected to system bus 806. Network interface unit 818 can also connect to other types of communication networks and devices, including connections via Bluetooth, Wi-Fi, and cellular telecommunications networks (including 4G and 5G networks). Network interface unit 818 can connect the control unit 40 to other networks, systems, and devices. The control unit 40 also includes an input / output unit 822 for receiving and processing inputs and outputs from peripheral devices.

[0096] Mass storage device 812 and RAM 808 can store software instructions and data. The software instructions may include an operating system 814 suitable for controlling the operation of sample processing instrument 1. Mass storage device 812 and / or RAM 808 may also store software instructions and data 816, which, when executed by processing device 802, provide the functionality of sample processing instrument 1 discussed herein.

[0097] Illustrative examples of the systems and methods described herein are provided below. Embodiments of the systems or methods described herein may include any one or more of the terms described below, and any combination thereof.

[0098] Clause 1. A method for operating a sample processing instrument, the method comprising: processing one or more samples via a flow control system of the sample processing instrument; cleaning the flow control system; measuring a particle quantity at a detection point of the flow control system by selectively supplying a flow from one of a sample tube source and a sample tubing source to the flow control system, and measuring the particle quantity in the flow; comparing the measured particle quantity with a predetermined target value; and using the comparison to determine whether a cleaning requirement is met.

[0099] Clause 2. The method according to Clause 1, wherein the flow is supplied from the sample tubing source and it is determined that the cleaning requirement is not met, the method further comprising: supplying a second flow from the sample tubing source; measuring a second particle amount in the second flow; comparing the measured second particle amount with the predetermined target value; and using the comparison to determine whether the cleaning requirement is met.

[0100] Clause 3. The method according to Clause 1 or 2, wherein the source of the stream is determined based on user input.

[0101] Clause 4. The method according to any one of Clauses 1 to 3, wherein the predetermined target value differs based on whether the flow is supplied from the sample tube source or from the sample line source.

[0102] Clause 5. The method according to Clause 4, wherein the predetermined target value is higher for the flow supplied from the sample tube source and lower for the flow supplied from the sample tubing source.

[0103] Clause 6. The method according to any one of Clauses 1 to 5, wherein the sample tubing source comprises a sheath fluid.

[0104] Clause 7. The method according to Clause 6, wherein the sheath fluid is a 5-nanometer sheath fluid.

[0105] Clause 8. The method according to any one of Clauses 1 to 7, wherein the sample tube source is a sample buffer.

[0106] Clause 9. The method according to any one of Clauses 1 to 8, wherein the predetermined target value is 100 events.

[0107] Clause 10. A system for operating a sample processing instrument, the system comprising: a flow control system including a flow chamber, a sample tube source, a sample tubing source, and a flow control path leading to the flow chamber; a source selection valve communicating with the flow control path and selectively communicating with each of the sample tube source and the sample tubing source; and a controller including a processor and a memory, the memory including instructions that, when executed by the processor, cause the processor to: generate a display including a source selection operation; receive input from the source selection operation; and operate the source selection valve using the input to provide flow from one of the sample tube source and the sample tubing source to the flow chamber.

[0108] Clause 11. The system according to Clause 10, wherein the source selection operation is associated with determining whether a cleanliness requirement is met based on the amount of particles measured at a detection point in the flow control system.

[0109] Clause 12. The system as described in Clause 11, wherein the cleaning requirement includes a predetermined threshold particle quantity.

[0110] Clause 13. The system according to Clause 12, wherein the predetermined threshold granularity is 100 events.

[0111] Clause 14. The system according to any one of Clauses 10 to 13, wherein the sample tubing source is a sheath fluid.

[0112] Clause 15. The system according to Clause 14, wherein the sheath fluid is a 5-nanometer sheath fluid.

[0113] Clause 16. The system according to any one of Clauses 10 to 15, wherein the sample processing instrument is a flow cytometer.

[0114] Clause 17. A system for operating a sample processing instrument, the system comprising: at least one processor; and a memory communicating with the processor and including instructions, which, when executed by the at least one processor, cause the processor to: determine whether a flow control system of the sample processing instrument meets a cleanliness requirement based on the amount of particles present in a measurement stream; and generate a graphical user interface including a source selection operation, wherein the source selection operation accepts input from a user selecting one of a sample tube source and a sample tubing source as the source of the measurement stream.

[0115] Clause 18. The system according to Clause 17, wherein the sample tube source is a sample buffer and the sample tubing source is a sheath fluid.

[0116] Clause 19. The system according to Clause 18, wherein the sheath fluid is a 5-nanometer sheath fluid.

[0117] Clause 20. The system according to Clause 18 or 19, wherein the cleaning requirement includes a threshold particle quantity, and the first threshold particle quantity of the sample tubing source is a smaller amount than the second threshold particle quantity of the sample tubing source.

[0118] Having described the preferred aspects and embodiments of this disclosure, modifications and equivalents to the disclosed concept will readily occur to those skilled in the art. However, such modifications and equivalents are intended to be included within the scope of the appended claims.

Claims

1. A method for operating a sample processing instrument, the method comprising: One or more samples are processed using the flow control system of the sample processing instrument; Clean the flow control system; The particle quantity at the detection point of the flow control system is measured using the following method: The flow control system can selectively supply flow from either a sample tube source or a sample tubing source. as well as Measure the amount of particles in the flow; The measured particle count is compared with a predetermined target value; as well as The comparison is used to determine whether the cleaning requirements are met.

2. The method of claim 1, wherein the flow is supplied from the sample tube source, and the cleaning requirement is determined not to be met, the method further comprising: A second stream is supplied from the sample tubing source; Measure the amount of the second particle in the second stream; The measured amount of the second particle is compared with the predetermined target value; as well as The comparison is used to determine whether the cleaning requirements are met.

3. The method of claim 1 or 2, wherein the source of the stream is determined based on user input.

4. The method according to any one of claims 1 to 3, wherein the predetermined target value differs based on whether the flow is supplied from the sample tube source or from the sample line source.

5. The method of claim 4, wherein the predetermined target value is higher for the flow supplied from the sample tube source and lower for the flow supplied from the sample line source.

6. The method according to any one of claims 1 to 5, wherein the sample tubing source comprises a sheath fluid.

7. The method according to claim 6, wherein the sheath fluid is a 5-nanometer sheath fluid.

8. The method according to any one of claims 1 to 7, wherein the predetermined target value is 100 events.

9. A system for operating a sample processing instrument, the system comprising: A flow control system, comprising: Flow chamber; Sample tube source; Sample tubing source; The flow control path leading to the flow chamber; A source selection valve, which is connected to the flow control path and selectively connected to each of the sample tube source and the sample tubing source; as well as The controller includes a processor and a memory, the memory including instructions that, when executed by the processor, cause the processor to: Generate a display that includes source selection operations; Receive input from the source selection operation; and The input is used to operate the source selection valve to supply flow from either the sample tube source or the sample tubing source to the flow chamber.

10. The system of claim 9, wherein the source selection operation is associated with determining whether a cleanliness requirement is met based on the particle quantity measured at a detection point in the flow control system.

11. The system of claim 10, wherein the cleaning requirement includes a predetermined threshold particle quantity.

12. The system of claim 11, wherein the predetermined threshold particle size is 100 events.

13. The system according to any one of claims 10 to 12, wherein the sample tubing source is a sheath fluid.

14. The system of claim 13, wherein the sheath fluid is a 5-nanometer sheath fluid.

15. The system according to any one of claims 10 to 14, wherein the sample processing instrument is a flow cytometer.

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