Apparatus for supplying reagents to a flow cytometry system

The disclosure provides example rinse station apparatus, cartridges, and methods for flow cytometry. The rinse station apparatus includes: (a) a cartridge docking station having a base with a recessed receptacle for a cartridge, a vertical support coupled to the base's first end and a top support coupled to the vertical support and cantilevered over the base, the top support has an opening that aligns with the cartridge's opening, (b) a locking arm coupled to the base's second end, the locking arm's free end has a ridge to cooperate with a detent coupled to the cartridge's rear wall to retain the cartridge in place, (c) a spring coupled to the vertical support's front face to apply force to the cartridge's front wall to bias the cartridge toward the locking arm, and (d) a load cell coupled to the base of the cartridge docking station, the load cell measure's the cartridge's weight.

BACKGROUND

In flow cytometry, streams of cells or particles suspended in fluid are passed in front of laser beam(s). Detectors capture the way the laser light is reflected and scattered off each particle to measure its physical properties. Often this requires sampling from fluids where the particles are not dissolved and will fallout of suspension over time due to gravity. The device which samples the fluid will be unable to reach all the particles once they fallout of suspension. The fluids are usually contained in vials, assay plates, or consumable cartridges.

For fluids that are sampled from over a long duration or are left stationary for long periods prior to sampling this is problematic. For example, if the fluid is sampled from for over a few hours, the number of events passing through the flow cytometer will decrease over time as the particles fall out of suspension. This runs the risk of failed experiments and potential loss of expensive assays and reagents. The current solution is to have the user periodically agitate the cartridge or vial every few hours to re-suspend the particles. This presents an issue if the user is required to stop or pause an experiment to do so.

In particular, the iQue® PLUS is a flow cytometry system that measures cells and/or beads in liquid suspension from 96,384, 1536-well microplates. To do this, the flow cytometry system uses a metal probe connected to plastic tubing to pump the samples from the microplate to the cytometer for measurement. The pump runs continuously while the cytometer is measuring and while the probe moves from liquid sample to liquid sample in the microplate. As the probe moves between samples, air is pulled into the tubing and that air keeps the different samples separated in the tubing. The ForeCyt software looks at the stream of data received from the cytometer and virtually separates and identifies the different samples (called well Identification or Well ID).

The IntelliCyt air-gap sampling method can be confused if there are no detectable analytes in a sample. To solve this problem, extra samples which have fluorescent beads (called “marker beads”) can be introduced into the sample stream around each of the samples coming from the plate. The marker beads are fluorescently dyed polystyrene microspheres mixed in liquid buffer in a marker cartridge. Marker beads are used to improve Well ID in situations where there may not be enough data for the software to correct identify and virtually separate the samples. During sampling, the probe aspirates from the marker cartridge, then from the microplate, then again from the marker cartridge, and repeats. This technique surrounds the microplate samples with marker bead “sips” which the flow cytometry system can identify and use for Well ID.

In addition, known cartridges that hold various liquids for the flow cytometry system are not resealable, permit liquid and samples to spill or splash during loading in the flow cytometry system, and are difficult to load and unload from the flow cytometry system.

SUMMARY

In a first aspect, an example rinse station apparatus for flow cytometry is disclosed. The rinse station apparatus includes (a) at least one cartridge docking station having a base with a recessed receptacle configured to receive a cartridge, a vertical support coupled to a first end of the base and a top support coupled to the vertical support and cantilevered over the base, wherein the top support has an opening arranged therethrough that is configured to align with an opening in the cartridge; (b) a locking arm coupled to a second end of the base, a free end of the locking arm has a ridge configured to cooperate with a detent coupled to a rear wall of the cartridge to retain the cartridge in place on the at least one cartridge docking station; (c) a spring coupled to a front face of the vertical support and configured to apply force to a front wall of the cartridge to bias the cartridge toward the locking arm; and (d) at least one load cell coupled to the base of the at least one cartridge docking station, wherein the load cell is configured to measure a weight of the cartridge.

In a second aspect, an example cartridge for flow cytometry is disclosed. The cartridge includes (a) a housing having a top surface, a bottom surface, a pair of opposing sidewalls, a front wall and a rear wall that together define a cavity, wherein an opening is defined through the top surface of the housing adjacent to the front wall, and the opening is surrounded by an annular ring having a shoulder at a first end and a second end that extends into the cavity; and (b) a re-sealable plug having a tubular body with a flange arranged at a first end such that the tubular body is disposed within the annular ring and the flange abuts the shoulder of the annular ring, a cap is coupled to the flange of the re-sealable plug via a living hinge and is configured to move between a sealed position in which a portion of the cap is recessed within an opening of the re-sealable plug and an unsealed position in which the cap and living hinge extend over a portion of the top surface.

In a third aspect, an example method for using a rinse station apparatus for flow cytometry is disclosed. The method includes (a) removably coupling the at least one cartridge according to the second aspect with the rinse station apparatus according to the first aspect; (b) determining, via the at least one load cell, a weight of the at least one cartridge and contents thereof; (c) receiving, via a microcontroller, a signal that includes a load cell value corresponding to the weight of the at least one cartridge; (d) in response to receiving the signal that includes the load cell value corresponding to the weight of the at least one cartridge, transmitting, via the microcontroller, a signal to an embedded processor that controls a motorized carrier coupled to a probe for sampling within the at least one cartridge; and (e) determining, via the embedded processor, a depth of the contents of the at least one cartridge based on the load cell value.

In a fourth aspect, an example non-transitory computer-readable medium is disclosed. The computer readable medium has stored thereon program instructions that upon execution by a processor, cause performance of a set of acts including (a) at least one load cell of the rinse station apparatus according to the first aspect determining a weight of at least one cartridge according to the second aspect and contents thereof; (b) a microcontroller receiving a signal that includes a load cell value corresponding to the weight of the at least one cartridge; (c) in response to receiving the signal that includes the load cell value corresponding to the weight of the at least one cartridge, the microcontroller transmitting a signal to an embedded processor that controls a motorized carrier coupled to a probe for sampling within the at least one cartridge; and (d) the embedded processor determining a depth of the contents of the at least one cartridge based on the load cell value.

The drawings are for the purpose of illustrating examples, but it is understood that the inventions are not limited to the arrangements and instrumentalities shown in the drawings.

DETAILED DESCRIPTION

Embodiments of the rinse station apparatus, cartridge and methods described herein can be used to determine a weight of the cartridge and contents thereof, determine a depth of the contents of the cartridge and determine a sampling depth for a tip of a probe within the cartridge. The disclosed example rinse station apparatus, cartridge and methods also beneficially enable control of local vortexing microfuge shaker, vibration motor and/or a linear actuator to place marker beads or cells in suspension.

II. Example Architecture

FIG. 1is a block diagram showing an operating environment100that includes or involves, for example, a rinse station apparatus105and at least one cartridge110shown in detail inFIGS. 3-11and described below. Method300inFIG. 12described below shows an embodiment of a method that can be implemented within this operating environment100.

FIG. 2is a block diagram illustrating an example of a computing device200, according to an example implementation, that is configured to interface with operating environment100, either directly or indirectly. The computing device200may be used to perform functions of the method shown inFIG. 12and described below. In particular, computing device200can be configured to perform one or more functions, including determining a depth of the contents of the at least one cartridge based on the load cell value and determining a sampling depth for a tip of the probe within the cartridge, for example. The computing device200has a processor(s)202, and also a communication interface204, data storage206, an output interface208, and a display210each connected to a communication bus212. The computing device200may also include hardware to enable communication within the computing device200and between the computing device200and other devices (e.g. not shown). The hardware may include transmitters, receivers, and antennas, for example.

The communication interface204may be a wireless interface and/or one or more wired interfaces that allow for both short-range communication and long-range communication to one or more networks214or to one or more remote computing devices216(e.g., a tablet216a, a personal computer216b, a laptop computer216cand a mobile computing device216d, for example). Such wireless interfaces may provide for communication under one or more wireless communication protocols, such as Bluetooth, WiFi (e.g., an institute of electrical and electronic engineers (IEEE) 802.11 protocol), Long-Term Evolution (LTE), cellular communications, near-field communication (NFC), and/or other wireless communication protocols. Such wired interfaces may include Ethernet interface, a Universal Serial Bus (USB) interface, or similar interface to communicate via a wire, a twisted pair of wires, a coaxial cable, an optical link, a fiber-optic link, or other physical connection to a wired network. Thus, the communication interface204may be configured to receive input data from one or more devices, and may also be configured to send output data to other devices.

The communication interface204may also include a user-input device, such as a keyboard, a keypad, a touch screen, a touch pad, a computer mouse, a track ball and/or other similar devices, for example.

The data storage206may include or take the form of one or more computer-readable storage media that can be read or accessed by the processor(s)202. The computer-readable storage media can include volatile and/or non-volatile storage components, such as optical, magnetic, organic or other memory or disc storage, which can be integrated in whole or in part with the processor(s)202. The data storage206is considered non-transitory computer readable media. In some examples, the data storage206can be implemented using a single physical device (e.g., one optical, magnetic, organic or other memory or disc storage unit), while in other examples, the data storage206can be implemented using two or more physical devices.

The data storage206thus is a non-transitory computer readable storage medium, and executable instructions218are stored thereon. The instructions218include computer executable code. When the instructions218are executed by the processor(s)202, the processor(s)202are caused to perform functions. Such functions include, but are not limited to, determining a weight of the cartridge and contents thereof and determining a depth of the contents of the at least one cartridge based on the load cell value.

The processor(s)202may be a general-purpose processor or a special purpose processor (e.g., digital signal processors, application specific integrated circuits, etc.). The processor(s)202may receive inputs from the communication interface204, and process the inputs to generate outputs that are stored in the data storage206and output to the display210. The processor(s)202can be configured to execute the executable instructions218(e.g., computer-readable program instructions) that are stored in the data storage206and are executable to provide the functionality of the computing device200described herein.

The output interface208outputs information to the display210or to other components as well. Thus, the output interface208may be similar to the communication interface204and can be a wireless interface (e.g., transmitter) or a wired interface as well. The output interface208may send commands to one or more controllable devices, for example

The computing device200shown inFIG. 2may also be representative of a local computing device200ain operating environment100, for example, in communication with rinse station apparatus105. This local computing device200amay perform one or more of the steps of the method300described below, may receive input from a user and/or may send image data and user input to computing device200to perform all or some of the steps of method300. In addition, in one optional example embodiment, the iQue® PLUS flow cytometry platform may be utilized to perform method300and includes the combined functionality of computing device200and rinse station apparatus105.

FIG. 12shows a flowchart of an example method300to determining, via the embedded processor, a depth of the contents of the at least one cartridge based on the load cell value, according to an example implementation. Method300shown inFIG. 12presents an example of a method that could be used with the computing device200ofFIG. 2, for example. Further, devices or systems may be used or configured to perform logical functions presented inFIG. 12, such as the microcontroller, embedded processor and workstation computer shown inFIG. 11. In some instances, components of the devices and/or systems may be configured to perform the functions such that the components are configured and structured with hardware and/or software to enable such performance. Components of the devices and/or systems may be arranged to be adapted to, capable of, or suited for performing the functions, such as when operated in a specific manner. Method300may include one or more operations, functions, or actions as illustrated by one or more of blocks305-325. Although the blocks are illustrated in a sequential order, some of these blocks may also be performed in parallel, and/or in a different order than those described herein. Also, the various blocks may be combined into fewer blocks, divided into additional blocks, and/or removed based upon the desired implementation.

It should be understood that for this and other processes and methods disclosed herein, flowcharts show functionality and operation of one possible implementation of the present examples. In this regard, each block may represent a module, a segment, or a portion of program code, which includes one or more instructions executable by a processor for implementing specific logical functions or steps in the process. The program code may be stored on any type of computer readable medium or data storage, for example, such as a storage device including a disk or hard drive. Further, the program code can be encoded on a computer-readable storage media in a machine-readable format, or on other non-transitory media or articles of manufacture. The computer readable medium may include non-transitory computer readable medium or memory, for example, such as computer-readable media that stores data for short periods of time such as register memory, processor cache and Random Access Memory (RAM). The computer readable medium may also include non-transitory media, such as secondary or persistent long term storage, like read only memory (ROM), optical or magnetic disks, compact-disc read only memory (CD-ROM), for example. The computer readable media may also be any other volatile or non-volatile storage systems. The computer readable medium may be considered a tangible computer readable storage medium, for example.

In addition, each block inFIG. 12, and within other processes and methods disclosed herein, may represent circuitry that is wired to perform the specific logical functions in the process. Alternative implementations are included within the scope of the examples of the present disclosure in which functions may be executed out of order from that shown or discussed, including substantially concurrent or in reverse order, depending on the functionality involved, as would be understood by those reasonably skilled in the art.

III. Example Rinse Station Apparatus

In a first aspect, shown inFIGS. 3-11, a rinse station apparatus105for flow cytometry includes at least one cartridge docking station115having a base120with a recessed receptacle125configured to receive a cartridge110, a vertical support130coupled to a first end121of the base120and a top support135coupled to the vertical support130and cantilevered over the base120. The top support135has an opening136arranged therethrough that is configured to align with an opening147in the cartridge110. In one optional embodiment, a surface126of the recessed receptacle125that is configured to support the cartridge110has an angle ranging from 1 degree to 45 degrees such that the surface126of the recessed receptacle125is inclined from the first end121of the base120toward the second end122of the base120.

In one optional embodiment, the rinse station apparatus105includes at least one cartridge110removably coupled to the at least one cartridge docking station115. The cartridge110includes a housing140having a top surface141, a bottom surface142, a pair of opposing sidewalls143, a front wall144and a rear wall145that together define a cavity146. An opening147is defined through the top surface141of the housing140adjacent to the front wall144. The opening147is surrounded by an annular ring148having a shoulder149at a first end150and a second end151that extends into the cavity146. And the cartridge110includes a re-sealable plug155having a tubular body with a flange156arranged at a first end157such that the tubular body158is disposed within the annular ring148and the flange156abuts the shoulder149of the annular ring148. A cap160is coupled to the flange156of the re-sealable plug155via a living hinge161and is configured to move between a sealed position in which a portion of the cap160is recessed within an opening163of the re-sealable plug155and an unsealed position in which the cap160and living hinge161extend over a portion of the top surface141. In alternative embodiments, a screw-on cap or cork-like plug may be used in place of the re-sealable plug155. One example cartridge110has a maximum fill volume of 51 ml, a normal fill volume of 42 ml, and a dead volume of 5 ml. The example cartridge110may be made of polypropylene with 3% white colorant. In addition, the example cartridge110may be made from two injection molded parts that are ultrasonically welded together. The re-sealable plug155may then be press-fit into place in the opening147of the top surface141.

In one optional embodiment, the cartridge110includes a cone-shaped baffle165coupled to the second end151of the annular ring148. The technical effect of the cone-shaped baffle165is to minimize any splashing of the contents of the cartridge110out of the opening147by dampening the liquid movement around the re-sealable plug155. In alternative embodiments, the baffle may take the form of a rim or plate that extends radially inward from the second end151of the annular ring148thereby reducing the diameter of the opening147at the second end151.

In another optional embodiment, the cartridge110includes a protrusion165coupled to and extending from the rear wall145of the housing140adjacent to the top surface141of the housing140. In operation, when the cartridges110are installed on the cartridge docking station115, the cartridges110are in a closely spaced array. As a result, cartridge110accessibility for handling is reduced. The protrusion165acts as a grip that is configured to be graspable between a finger and thumb of an operator. The protrusion165may optionally include ridges166on the top and bottom sides to improve grip. The ridges166may also provide a visual cue to the operator as to the location to grasp the protrusion165. The ridges166are curved on the top surface for aesthetics.

In yet another optional embodiment, the cartridge110includes a detent170coupled to the rear wall145of the housing140adjacent to the bottom surface142of the housing140. In this example, the detent170acts as heel that locks into place with the locking arm175described below.

In still another optional embodiment, shown inFIG. 8, the shoulder149of the annular ring148is inset in the top surface141of the housing140. In another optional embodiment, shown inFIG. 9, the shoulder149of the annular ring148corresponds to the first end150of the annular ring148that extends above the top surface141of the housing140. In a further optional embodiment, the bottom surface142of the housing140has an angle ranging from 1 degree to 45 degrees such that the cartridge110is inclined from the front wall144toward the rear wall145. The technical effect of such an arrangement is to pool the liquid contents of the cartridge110under the opening147as the volume of the contents decreases due to sampling.

The rinse station apparatus105also includes a locking arm175coupled to a second end122of the base120. A free end176of the locking arm175has a ridge177configured to cooperate with a detent coupled to a rear wall of the cartridge110to retain the cartridge110in place on the at least one cartridge docking station115. In one optional embodiment, the locking arm175is biased toward a locked position and is configured to flex outwardly to an open position under application of a downward force or an upward force from the detent170of the cartridge110.

The rinse station apparatus105further includes a spring180coupled to a front face131of the vertical support130and configured to apply force to the front wall144of the cartridge110to bias the cartridge110toward the locking arm175in the recessed receptacle125. The technical effect of the spring180is to help retain the cartridge110in place on the cartridge docking station115, in example embodiments, the optional detent170of the cartridge110is located under the ridge177of the locking arm175. And when an operator grips the optional protrusion165of the cartridge110and applies an upward force, the spring180applies a force to the front wall144of the cartridge advancing the cartridge toward the second end122of the base120thereby increasing the ease of removal of the cartridge110from the cartridge docking station115.

The rinse station apparatus105also includes at least one load cell185coupled to the base120of the at least one cartridge docking station115. The load cell185is configured to measure a weight of the cartridge110and the contents therein. The technical effect of the load cell185and the corresponding weight value is to facilitate a determination of a sampling depth of a tip109of a probe103within the cartridge110and to facilitate a determination and alert for an operator indicating when the cartridge is low or empty or not loaded on the cartridge docking station115. The load cell185and the corresponding weight value may also be used to calculate run-time for sampling or determine the number of well-plates that remain for sampling. Still further, alerts may be generated based on user-defined limits or thresholds that are preset on the system.

In one optional embodiment, the at least one load cell185has a cross-arm186with fixed-geometry stop187configured to limit displacement of the load cell185in response to a docking force from the cartridge110. In one optional embodiment, shown inFIG. 5, the fixed-geometry stop187has the form of a keyed cut out dividing the cross-arm186into a first portion188and a second portion189that overlap such that the first portion188of the cross-arm186is configured to flex downward in response to a force from the cartridge110during docking until the first portion188of the cross-arm186contacts the second portion189of the cross-arm186at the keyed cut out. As used herein, “keyed cut out” refers to a gap between the first and second portions of the cross-arm of a load cell where the gap is shaped to provide a reciprocal male-female arrangement between the first and second portions such that these first and second portions of the cross-arm overlap and are “keyed” together. The fixed-geometry stop187may take many forms that prevent the load cell185from displacing too far and damaging the load cell's strain gauge. For example, a lockable adjustment screw, shims or other types of machined gaps may be coupled to the load cell to contact the base120upon displacement and to set a predetermined displacement distance. In one example, the maximum displacement is about 0.3 mm.

In one optional embodiment, the rinse station apparatus105includes a vortexing microfuge shaker195having a motor196coupled to a first end182of a carrier vessel197for a microfuge tube172via a spherical joint198, a platform199suspended above the motor196via a plurality of uprights181, the platform199supporting a second end183of the carrier vessel197, the platform199has a cap retention cavity184arranged adjacent to the second end183of the carrier vessel197and that is configured to receive a cap171coupled to the microfuge tube172via a living hinge173. The motor may take the form of an eccentric gear motor or DC motor. In one embodiment, the spherical joint198is coupled to the motor196via an off-center coupling to the motor shaft thereby imparting eccentric motion to the carrier vessel197. In operation, the rinse station apparatus105can aspirate samples from the microfuge tube172for analysis. For example, the microfuge tube172can contain samples for analysis that are typically either beads or cells. The microfuge tube172may also contain quality control beads that have specific fluorescent properties and that allow the rinse station apparatus105to self-check a flow cytometer to confirm whether the flow cytometer is measuring within specifications.

The eccentric gear motor196is configured to rotate the first end182of the carrier vessel197off-axis. That rotation at the first end182causes three dimensional rotation about spherical joint198at the second end183of the carrier vessel197. The rotation causes liquid in the microfuge tube172to move up the side of the microfuge tube172and slide along the side as it rotates which causes mixing. Experiments have shown that the vortexing microfuge shaker195is effective at maintaining bead and cell suspension in the liquid and can re-suspend within 15 seconds of powering on the eccentric gear motor196.

In one optional embodiment, the at least one cartridge docking station115is a plurality of cartridge docking stations115arranged adjacent to each other, and the at least one load cell185is a plurality of load cells185each coupled to the base120of one of the plurality of cartridge docking stations115. These cartridges110may include reagents like decontamination solution, cleaning solution, buffer solution, rinse water, and marker bead solutions. The reagents can be aspirated by the probe103and provide different functions. One function is to provide a buffer solution which keeps the correct air-liquid ratios in the tubing when the probe103is temporarily not aspirating samples from the well plate106. Another function is to provide cleaning solutions to the probe103and tubing which rinse or dissolve away contaminates.

In one optional embodiment, the rinse station apparatus105includes a unifying housing132having a top surface133configured to overlie the top support135of each of the plurality of cartridge docking stations115and the platform199of the vortexing microfuge shaker195. The top surface133of the unifying housing132has a plurality of openings134therethrough each aligned with one of the openings136of the top support135of the plurality of cartridge docking stations115, an opening of the carrier vessel197of the vortexing microfuge shaker195, and the cap retention cavity184. The unifying housing132has a first vertical support137coupled to the top surface133at a first end. The first vertical support137forms a cavity configured to receive the vortexing microfuge shaker195. The unifying housing132has a second vertical support138coupled to the top surface133at a second end. And the unifying housing132has a vertical wall139extending between the first vertical support137and the second vertical support138that together form a recess127to receive the first end121of the base120, the vertical support130and the top support135of each of the plurality of cartridge docking stations115. And the unifying housing132has a base frame128extending outward from the vertical wall139with an opening129configured to surround the plurality of cartridge docking stations115and the plurality of load cells185. A height of the base frame128corresponds to a combined height of one of the plurality of load cells185coupled to one of the plurality of cartridge docking stations115.

In one optional embodiment, shown inFIG. 11, the rinse station apparatus105includes a fluid station circuit board101electrically coupled to the at least one load cell185. As used herein, “electrically coupled” refers to coupling using a conductor, such as a wire or a conductible trace, as well as inductive, magnetic and wireless couplings. The rinse station apparatus105also includes an analog-to-digital converter electrically coupled to the fluid station circuit board101and the at least one load cell185. The rinse station apparatus105further includes a microcontroller102electrically coupled to the fluid station circuit board101. And the rinse station apparatus105includes a probe103coupled to a motorized carrier104that is electrically coupled to the microcontroller102. The motorized carrier104is configured to move between the at least one cartridge docking station115and a well plate106. The probe103has an outer support sleeve107and an inner probe108that extends a distance beyond the outer support sleeve107. The outer support sleeve107provides mechanical support to the inner probe108. Yet a small gap exists between the outer support sleeve107and the inner probe108that may permit wicking and therefore contamination. As such, the microcontroller102is configured to receive a signal that includes a load cell value corresponding to a weight of the cartridge110and to transmit a signal to the motorized carrier104that includes a sampling depth for a tip109of the probe103within the cartridge110. This has the advantage of preventing wicking action and avoiding contamination. In alternative embodiments, the signal that includes a sampling depth for a tip109of the probe103within the cartridge110is transmitted to the motorized carrier104via another processor in the operating environment100such as an embedded processor111or workstation computer112.

In one optional embodiment, the vortexing microfuge shaker195is electrically coupled to the fluid station circuit board101and the microcontroller102is configured to transmit a signal to the eccentric gear motor196of the vortexing microfuge shaker195via the fluid station circuit board101to power on, to power off or to power on for a specified duration.

In one optional embodiment, as shown inFIG. 5, the base120of the at least one cartridge docking station115is coupled to the at least one load cell185such that the first end121of the base120is cantilevered off of a first end190of the load cell185. In one optional embodiment, the base120of the at least one cartridge docking station115is coupled to a first end190of the at least one load cell185via a pivot mount191such that the first end121of the base120is cantilevered off of the first end190of the load cell185and the base120is elevated above the at least one load cell185. In a further optional embodiment, the rinse station apparatus105includes a pair of flexible supports192having a first end193coupled to the base120and a second end194coupled to the load cell185. The pair of flexible supports192are arranged on either side of the pivot mount191and are configured to provide a restorative force to the base120in response to being compressed. These flexible supports192may take the form of leaf springs, torsion springs, compression springs or any other mechanism that is configured to provide a restorative force. The rinse station apparatus105also includes a vibration motor113coupled to a bottom side of the first end121of the base120of the at least one cartridge docking station115and configured to impart a rocking motion to the at least one cartridge docking station115about the pivot mount191. In one optional embodiment, the fluid station circuit board101is electrically coupled to the vibration motor113and the microcontroller102is configured to transmit a signal to the vibration motor113via the fluid station circuit board101to power on, to power off or to power on for a specified duration. The probe103is able to sample from the cartridge110during vibration.

To demonstrate the effectiveness of the foregoing example embodiment, four tests were conducted with a prototype cartridge docking station115coupled to a vibration motor113. Two tests were conducted with the vibration motor113in operation and the other two tests were conducted with the vibration motor113powered off. For each test run, a 29 mL cartridge110was filled with a solution containing marker beads. The cartridge110was sampled once a minute over the course of 25 hours. The samples were passed through a flow cytometer system to count the number of marker beads in each sample.

The number of marker beads decreased linearly over the 25 hours for the two test runs without any agitation by the vibration motor113, and the bead counts approached zero. This demonstrated the marker beads were falling out of suspension and settling. The two test runs with agitation by the vibration motor113did not show a linear decrease in marker bead counts. Instead, the marker bead counts rapidly decreased for the first few hours until the system reached steady state between 100-120 particles per sample. Then the marker bead counts and concentration plateaued and remained constant for the remainder of the experiment.

For purposes of operation, the particle concentration does not need to remain at a consistent level. Rather, the concentration should remain above a certain threshold to prevent Well-ID from failing. The number of particles in the fluid of cartridge110can be increased by the vibration motor113to prevent the steady state particle concentration from falling below the minimum threshold. The duration of agitation by the vibration motor113and the amplitude may be increased to achieve the same result. The amplitude and duration of agitation may also be adjusted based on the liquid-level measurements received from the load cell185.

In one optional embodiment, the rinse station apparatus105includes a linear actuator114fixedly coupled to a first end190or the second end179of the at least one load cell185, the linear actuator114having an actuating arm116coupled to the first end121or the second end122of the base120of the at least one cartridge docking station115and configured to impart a rocking motion to the at least one cartridge docking station115about the pivot mount191. In an alternative embodiment, the linear actuator114may have an actuating arm116coupled to the second end122of the base120of the at least one cartridge docking station115. In one optional embodiment, the fluid station circuit board101is electrically coupled to the linear actuator114and the microcontroller102is configured to transmit a signal to the linear actuator114via the fluid station circuit board101to power on, to power off or to power on for a specified duration. In addition, the amplitude and frequency of the linear actuator114may be adjusted based on the liquid-level measurements received from the load cell185. The probe103is able to sample from the cartridge110during linear actuation.

In operation, the technical effect of the vibration motor113and the linear actuator is to shake the cartridge110to avoid the operator having to manually re-suspend the fluid in the cartridge110. Activation of the vibration motor113or linear actuator114causes cartridge docking station115to vibrate or rock and thereby promotes mixing of the fluid in the cartridge110. The vibration motor113and the linear actuator114may beneficially permit long duration testing to be conducted without the need to pause or stop an experiment and re-suspend the fluid, faster start up on future experiments, improved consistency in particle concentration within the cartridge110, prevent particles from adhering to the walls of the cartridge110and permit adjustments for multiple different fluid and particle solutions.

In one optional embodiment, the rinse station apparatus105includes a shield circuit board117electrically coupled to the microcontroller102. In operation, the shield circuit board117steps down the voltage to appropriately interface with the rinse station apparatus105and various motors included therein. The shield circuit board117also includes relays that control the motors of the rinse station apparatus105. The rinse station apparatus105also includes an embedded processor111electrically coupled to the microcontroller102and to the shield circuit board117. And the rinse station apparatus105further includes a workstation computer112electrically coupled to the embedded processor111and configured to receive and process commands from an operator. In a further optional embodiment, a USB Hub118may be electrically coupled to a power supply119, to the embedded processor111and to the microcontroller102in order to provide power to the embedded processor111and the microcontroller102.

In still another optional embodiment, the rinse station apparatus105includes reciprocal mating components between the at least one cartridge110and the at least one cartridge docking station115. The reciprocal mating components include (i) at least one male component extending from either the base120of the at least one cartridge docking station115or the bottom surface142of the housing140of the at least one cartridge110and at least one corresponding female component defined within the other of the base120of the at least one cartridge docking station115or the bottom surface142of the housing140of the at least one cartridge110or (ii) a first RFID tag coupled to the at least one cartridge docking station115configured to pair with a second RFID tag coupled to the at least one cartridge110and to send a signal to the microcontroller102to indicate a match, (iii) a barcode coupled to the at least one cartridge and a scanner coupled to the at least one cartridge docking station, or (iv) a QR code coupled to the at least one cartridge and a camera and processor with imaging software coupled to the at least one cartridge docking station. The technical effect of the reciprocal mating components is to assist the operator with placement of the cartridge100with the correct liquid contents in the corresponding cartridge docking station115.

IV. Example Methods

Referring now toFIG. 12, a method300is illustrated using the rinse station apparatus105ofFIGS. 3-11and computing device ofFIGS. 1-2. Method300includes, at block305, removably coupling the at least one cartridge110according to any one of foregoing embodiments with the rinse station apparatus105according to any one of the foregoing embodiments. Then, at block310, the at least one load cell185determines a weight of the at least one cartridge110and contents thereof. Next, at block315, the microcontroller102receives a signal that includes a load cell value corresponding to the weight of the at least one cartridge110. In alternative embodiments, the signal that includes the load cell value may be received by the embedded processor111or workstation computer112. Then, in response to receiving the signal that includes the load cell value corresponding to the weight of the at least one cartridge110, the microcontroller102transmits a signal to an embedded processor111that controls a motorized carrier104coupled to a probe103for sampling within the at least one cartridge110, at block320. At block325, either the microcontroller102or the embedded processor111determines a depth of the contents of the at least one cartridge110based on the load cell value.

In one optional embodiment, the embedded processor111or the microcontroller102determines a sampling depth for a tip109of the probe103within the at least one cartridge110based on the determined depth of the contents of the at least one cartridge110. In operation, the sampling depth is approximately set for 8 mm of immersion to prevent tolerance stackup from causing the probe103to miss the liquid contents of the cartridge110. The tolerances are determined based on probe positioning, mechanical sensing of the cartridge location, and load cell measurements corresponding to the level of the liquid contents therein. The sampling depth is also calculated to maintain the probe103at least 0.5 mm from the bottom surface142of the cartridge110.

In one optional embodiment, the microcontroller102transmits a signal to an eccentric gear motor196of a vortexing microfuge shaker195to power on, to power off or to power on for a specified duration the vortexing microfuge shaker195.

In one optional embodiment, the method300further includes the microcontroller102transmitting a signal to a vibration motor113coupled to the base120of the at least one cartridge docking station115to power on, to power off or to power on for a specified duration. In an alternative optional embodiment, the method300includes the microcontroller102transmitting a signal to a linear actuator114coupled to the base120of the at least one cartridge docking station115to power on, to power off or to power on for a specified duration.

In one optional embodiment, the method300further includes the microcontroller102determining that the determined depth of the contents of the at least one cartridge110is below a sampling threshold, and the microcontroller102transmitting a signal to a workstation computer112to display an alert for an operator. The sampling threshold may be set at a depth that permits the system to continue operating for a set duration of time (e.g., several minutes) after the alert is displayed before the microcontroller102, the embedded processor111or the workstation computer112stop or pause the sampling activity. Alternatively, the sampling threshold may be set at a depth that immediately causes the microcontroller102, the embedded processor111or the workstation computer112stop or pause the sampling activity.

In one optional embodiment, the method300further includes applying a force, via an operator, to a protrusion coupled to and extending from the rear wall145of the housing140of the at least one cartridge110adjacent to the top surface141of the housing140of the at least one cartridge110and thereby flexing the locking arm175outwardly and releasing the detent170from the locking arm175. In this embodiment, method300also includes applying a force to a front wall144of the at least one cartridge110, via the spring180coupled to the front face131of the vertical support130of the at least one cartridge docking station115.

In one optional embodiment, the method300further includes converting, via an analog-to-digital converter electrically coupled to the fluid station circuit board101and the load cell185, an analog signal from the load cell185to a digital signal.

In one optional embodiment, a pair of reciprocal mating components are provided between the at least one cartridge110and the at least one cartridge docking station115. The reciprocal mating components comprise a first RFID tag coupled to the at least one cartridge docking station115configured to pair with a second RFID tag coupled to the at least one cartridge110and to send a signal to the microcontroller102to indicate a match. In this embodiment, method300further includes receiving, via the microcontroller102, a signal identifying the first RFID tag. Then, the microcontroller102receives a signal identifying the second RFID tag. Next, the microcontroller102determines whether the first RFID tag pairs with the second RFID tag. And the microcontroller102sends a signal with a determination of pairing between the first RFID tag and the second RFID tag.

In one optional embodiment, method300includes the embedded processor111determining the depth of the contents of a first cartridge110is at or below a minimum threshold vale based on the load cell value. And the embedded processor111sends a signal to the motorized carrier104to sample from a second cartridge110.

As discussed above, a non-transitory computer-readable medium having stored thereon program instructions that upon execution by a processor202may be utilized to cause performance of any of functions of the foregoing methods.

As one example, a non-transitory computer-readable medium having stored thereon program instructions that upon execution by a processor, cause performance of a set of acts includes at least one load cell185of the rinse station apparatus105according to any of the foregoing embodiments determining a weight of at least one cartridge110according to any of the foregoing embodiments and the contents thereof. A microcontroller102then receives a signal that includes a load cell value corresponding to the weight of the at least one cartridge110. In response to receiving the signal that includes the load cell value corresponding to the weight of the at least one cartridge110, the microcontroller102transmits a signal to an embedded processor111that controls a motorized carrier104coupled to a probe103for sampling within the at least one cartridge110. And the embedded processor111then determines a depth of the contents of the at least one cartridge110based on the load cell value.

In one optional embodiment, the non-transitory computer-readable medium further includes the embedded processor111determining a sampling depth for a tip109of the probe103within the at least one cartridge110based on the determined depth of the contents of the at least one cartridge110.

In another optional embodiment, the non-transitory computer-readable medium further includes the microcontroller102transmitting a signal to an eccentric gear motor196of a vortexing microfuge shaker195to power on, to power off or to power on for a specified duration the vortexing microfuge shaker195.

In a yet another optional embodiment, the non-transitory computer-readable medium further includes the microcontroller102transmitting a signal to a vibration motor113coupled to the base120of the at least one cartridge docking station115to power on, to power off or to power on for a specified duration. In an alternative optional embodiment, the microcontroller102transmits a signal to a linear actuator114coupled to the base120of the at least one cartridge docking station115to power on, to power off or to power on for a specified duration.

In still another optional embodiment, the non-transitory computer-readable medium further includes the microcontroller102determines that the determined depth of the contents of the at least one cartridge110is below a sampling threshold. And then the microcontroller102transmits a signal to a workstation computer112to display an alert for an operator.

In another optional embodiment, the non-transitory computer-readable medium further includes an analog-to-digital converter electrically coupled to the fluid station circuit board101and the load cell185converting an analog signal from the load cell185to a digital signal.

In yet another optional embodiment, a pair of reciprocal mating components are provided between the at least one cartridge110and the at least one cartridge docking station115. The reciprocal mating components comprise a first RFID tag coupled to the at least one cartridge docking station115configured to pair with a second RFID tag coupled to the at least one cartridge110and to send a signal to the microcontroller102to indicate a match. And the set of acts of the non-transitory computer-readable medium further includes the microcontroller102receiving a signal identifying the first RFID tag, the microcontroller102receiving a signal identifying the second RFID tag, the microcontroller102determining whether the first RFID tag pairs with the second RFID tag, and the microcontroller102sending a signal with a determination of pairing between the first RFID tag and the second RFID tag.

In a further optional embodiment, the non-transitory computer-readable medium further includes the embedded processor111determining the depth of the contents of a first cartridge110is at or below a minimum threshold vale based on the load cell value, and the embedded processor111sending a signal to the motorized carrier104to sample from a second cartridge110.