Patent Application
20170336427
Some embodiments described herein generally relate to online liquid autosamplers and processing systems for bacterial endotoxin monitoring.
Unless otherwise indicated herein, the materials described herein are not prior art to the claims in the present application and are not admitted to be prior art by inclusion in this section.
Bacterial endotoxins are fever producing materials that originate from the cell wall of gram negative bacteria and consist of lipopolysaccharide. Bacterial sepsis is a major cause of fatality worldwide. Sepsis is a multi-step process that involves an uncontrolled inflammatory response by the host cells that may result in multi organ failure and death. Both gram-negative and gram-positive bacteria play a major role in causing sepsis. These bacteria produce a range of virulence factors that enable them to escape the immune defenses and disseminate to remote organs, and toxins that interact with host cells via specific receptors on the cell surface and trigger a dysregulated immune response. Endotoxins make up about 75% of the outer membrane of gram-negative bacteria that are capable of causing lethal shock.
Pharmaceutical, biopharmaceutical, and medical device companies expend significant resources to detect, quantify, and eliminate endotoxins from their manufacturing environment and all product. Endotoxins may be detected using a test method known as Limulus Amebocyte Lysate (LAL) assay, which utilizes a reagent derived from amebocyte blood cells of the Atlantic Horseshoe Crab. The blood cells form a gelatinous clot in the presence of bacterial endotoxins and are used in various techniques of LAL testing.
The subject matter claimed herein is not limited to embodiments that solve any disadvantages or that operate only in environments such as those described above. Rather, this background is only provided to illustrate one exemplary technology area where some embodiments described herein may be practiced.
This Summary is provided to introduce a selection of concepts in a simplified form that are further described below in the Detailed Description. This Summary is not intended to identify key features or essential characteristics of the claimed subject matter, nor is it intended to be used as an aid in determining the scope of the claimed subject matter.
Some embodiments described herein generally relate to online liquid autosamplers and processing systems for bacterial endotoxin monitoring.
In an example embodiment, an online liquid autosampler may include a pipette, a depyrogenation system, a sample source, and a movement system. The depyrogenation system may be configured to selectively depyrogenate the pipette. The sample source may be configured to provide a liquid to be sampled to the pipette. The movement system may be coupled to one or more of the pipette, the depyrogenation system, or the sample source. The movement system may be configured to: position the pipette within the depyrogenation system for depyrogenation; position a tip of the pipette within the sample source to aspirate a sample of the liquid from the sample source; and position the pipette to dispense an aliquot of the sample of the liquid into an aliquot sample target.
In another example embodiment, a method to autosample a liquid for endotoxin analysis using an online liquid autosampler that includes a pipette, a depyrogenation system, a sample source, and a movement system is described. The method may include positioning the pipette within the depyrogenation system using the movement system. The method may also include depyrogenating the pipette within the depyrogenation system at least at a threshold temperature for at least a threshold amount of time. The method may also include positioning a tip of the pipette within the sample source using the movement system. The method may also include aspirating a sample of liquid from within the sample source using the pipette. The method may also include positioning the pipette above an aliquot sample target using the movement system. The method may also include dispensing an aliquot of the sample of the liquid into the aliquot sample target.
Additional features and advantages of the invention will be set forth in the description which follows, and in part will be obvious from the description, or may be learned by the practice of the invention. The features and advantages of the invention may be realized and obtained by means of the instruments and combinations particularly pointed out in the appended claims. These and other features of the present invention will become more fully apparent from the following description and appended claims, or may be learned by the practice of the invention as set forth hereinafter.
To further clarify the above and other advantages and features of the present invention, a more particular description of the invention will be rendered by reference to specific embodiments thereof which are illustrated in the appended drawings. It is appreciated that these drawings depict only typical embodiments of the invention and are therefore not to be considered limiting of its scope. The invention will be described and explained with additional specificity and detail through the use of the accompanying drawings in which:
all arranged in accordance with at least one embodiment described herein.
Endotoxin testing may be performed in compliance with the following guidelines: ANSI/AAMI ST72:2011; Bacterial Endotoxins-Test methods, routine monitoring, and alternatives to batch testing; USP <85> Bacterial Endotoxins Test Methods.
The Kinetic Chromogenic LAL method of testing is often used to quantify or detect endotoxins on the basis of a measured color producing reaction, proportional to the interaction of LAL and endotoxin. Samples are dispensed into microplates containing the LAL reagent that are read by a spectrophotometer. The spectrophotometer measures the color intensity directly related to endotoxin concentration of the sample.
Kinetic Chromogenic LAL methods have been in use within laboratories for decades. Handheld, portable methods have been introduced within the last decade and have gained acceptance globally for use in screening raw materials, intermediates, and final product.
Industries that are required to routinely monitor endotoxin levels are desirous of automated endotoxin monitoring systems such as described herein that able to be deployed online for automated periodic monitoring. Embodiments described herein may allow for scheduling of sampling events in the future. Some embodiments may execute these sampling events and capture test results for evaluation by personnel. Thus, some embodiments of automated online systems described herein may improve both the quality and economic impact of routine endotoxin monitoring.
Others have attempted but ultimately failed to satisfy the long felt, unmet need for automated online endotoxin monitoring systems. Lonza, Inc. (Basel, Switzerland) introduced their PyroSense® system in approximately 2008 which could be programmed to perform from 1 up to 12 automated assays per day. Lonza's PyroSense® system incorporated highly complex robotics, fluidics, wet reagents, and complex software algorithms to perform each test. Lonza's PyroSense® system is complicated, physically quite large and cumbersome, and quite expensive. The market determined this solution was not economically feasible.
Around 2006, Charles River Laboratories, Inc. (Waltham, Mass.) introduced a portable endotoxin testing system marketed as the Endosafe®-PTS™ (PTS1000). The company expanded the PTS product portfolio to include a multiple sample testing system known as the Endosafe®-MCS™ (multi-cartridge system) to satisfy the demand of its clients who require higher sample throughput. The MCS™ can test five cartridges simultaneously, or any combination from one to five. However, it is designed to operate within a controlled laboratory environment. It is not able to operate within the manufacturing environment nor support automated online operation with dynamic depyrogenation and management of continuously flowing samples.
Some embodiments described herein satisfy the longfelt global unmet need for online endotoxin monitoring solutions within pharmaceutical, biopharmaceutical, medical device, and other highly regulated markets.
Some embodiments disclosed herein include methods and apparatuses for automated sampling and processing of bacterial endotoxin monitoring of very small samples quantities at high accuracy. One of the critical requirements in performing bacterial endotoxin tests is that the sample container be pyrogen free. Depyrogenation typically is accomplished by heating the sample containers to 250° C. or higher for a period of at least 30 minutes. This depyrogenation routine must be completed prior to pipetting each sample within an online automated sampling and processing system. A particular disclosed online depyrogenation method retains the sampling pipette within a heating element enclosure or depyrogenation chamber. The internal temperature of the pipette is elevated to 250° C. or higher for a period of at least 30 minutes. The pipette is then allowed to cool to desired temperature prior to departing the depyrogenation chamber to collect the sample. The depyrogenation system may include the pipette, constructed of a specific material to withstand heating and cooling cycles, an enclosed heating element, insulation system, and controls system to bring the pipette to the required temperature for the required duration. The specific geometric shape of the pipette in combination with the specific alignment of the pipette tip to the sample receipt vessel may facilitate proper aspiration and dispensing of micro-fluidic samples.
Some embodiments of the sample processing system disclosed herein may ensure the sample product is retained within the pipette only during aspiration and delivery. This may prevent upstream tubing from becoming contaminated with sample product and may ensure proper depyrogenation of surfaces in contact with prior samples.
It is not typically desired for product being sampled to be stagnant as this may increase potential for contamination. It may be desired that the micro-sample be extracted from a flowing stream. A particular disclosed sampling well may facilitate the continuous flow of sample through a vessel (e.g., a sample well) which may allow sample accumulation and aspiration while not impeding flow rates. The combination of the pipette material and shape with the specific sampling well may provide accurate collection of micro-sample volumes from a continuously flowing input sample with input flow rates as low as 0.1 microliters per minute and higher.
In at least one embodiment, the combination of the pipette material and shape and the specific alignment of the pipette tip to the sample receipt vessel may provide accurate delivery of precise micro-volumes from 0.5 microliters up to 250 microliters. Accuracy of delivery may be dependent upon drop formation size which may be dependent upon fluid viscosity and surface tension. Some embodiments described herein may utilize the proximity of the delivery target to the pipette tip end to facilitate and/or ensure that the target delivery volume is consistently removed. The proximity between the target and tip may be variable in some embodiments and can be adjusted to compensate for changes in fluid properties as well as target and pipette properties (materials, dimension changes). In other embodiments higher sample volumes may be collected and delivered with precision and accuracy.
Embodiments of the electronic and embedded systems disclosed herein may enable the automated sampling and processing system disclosed to interface automatically with third parties' microfluidics cartridges and testing systems. In an example embodiment the automated sampling and processing system disclosed automatically separates a third party's single cartridge from a nested stack and then loads this cartridge into the third party's reader. The automated sampling and processing system disclosed may interface directly with the third party's reader device to initiate reader protocols at predetermined time intervals and extract digital copies of results from said third party's reader device. Other embodiments allow for interfacing with various microfluidic systems.
Embodiments of the automated sampling and processing system disclosed herein may manage scheduling of future sampling events. In one embodiment the operator simply inputs the day and time of desired sampling events and loads the microfluidics cartridges or other aliquot sample targets into the magazine. Embodiments of the automated sampling and processing systems disclosed herein may confirm availability of microfluidics cartridges required to complete scheduled events and may confirm the microfluidics cartridges are within their expiration dates per specifications provided by microcassette cartridge manufacturer.
Embodiments of the automated sampling and processing system disclosed herein may automatically initiate heating (e.g., depyrogenation) of the pipette and other required preparatory functions at specific timing intervals tied to the scheduled sampling event. At the scheduled event time the automated sampling and processing system may initiate depyrogenation, sample aspiration, loading of the microfluidics cartridge or other aliquot sample target into the reader, and delivery of precise sample volumes to one or more sample receipt vessels. Upon completion of the third party's reader activities, the automated sampling and processing system may automatically remove and discharge the used microfluidics cartridge and may await the next scheduled event. The automated sampling and processing systems described herein may alternatively or additionally be referred to herein as sample processing systems, online liquid autosamplers, autosamplers, or similar terms.
In an example embodiment, an online liquid autosampler may include a pipette, a depyrogenation system, a sample source such as a sample well, and a movement system. The depyrogenation system may be configured to selectively depyrogenate the pipette. The sample source may be configured to provide a liquid to be sampled to the pipette. The movement system may be coupled to one or more of the pipette, the depyrogenation system, or the sample source. The movement system may include a servo motor, a rotary actuator, a single axis actuator, a multi axis actuator, a robotic arm, a conveyor belt, or any other suitable devices or combinations thereof to move one or more elements relative to one or more other elements. For instance, the movement system may be configured to position the pipette within the depyrogenation system for depyrogenation, position a tip of the pipette within the sample source to aspirate a sample of the liquid from the sample source, and/or position the pipette to dispense an aliquot of the sample of the liquid into an aliquot sample target. The movement system may system may accomplish the foregoing in some embodiments by moving one or more of the pipette, the depyrogenation system, the sample source, and/or the aliquot sample target relative to one or more other components.
Alternatively or additionally, the online liquid autosampler may include a UV sanitizer, a rinse system, and/or other elements or feature as described herein.
Some embodiments may include and/or enable automated sampling and processing of bacterial endotoxin monitoring of very small sample quantities at high accuracy. An online depyrogenation method that may be implemented according to some embodiments may retain the pipette within a heating element enclosure of the depyrogenation system. The internal temperature of the pipette may be elevated to 250° C. or higher for a period of at least 30 minutes. The pipette may be constructed of a material to withstand heating and cooling cycles.
An example sample well according to some embodiments described herein may facilitate a continuous flow of the sample through a funnel or other vessel which may allow sample accumulation and aspiration without impeding flow rates. The pipette may provide accurate collection of micro-sample volumes from a continuously flowing input sample through the sample well with input flow rates as low as 0.1 microliters per minute and higher. In at least one embodiment, a combination of the pipette material and a shape of the pipette tip and the alignment of the pipette tip to the sample well may provide accurate delivery of precise micro-volumes from 0.5 microliters up to 250 microliters. Accuracy of delivery may be dependent upon drop formation size which may be dependent upon fluid viscosity and surface tension. Some embodiments described herein may utilize a proximity of the aliquot sample target to the pipette tip to ensure the target delivery volume is consistently removed from the pipette. The proximity (e.g., distance) between the aliquot sample target and the pipette tip may be adjusted to compensate for changes in fluid properties as well as aliquot sample target and pipette properties (materials, dimension changes). In other embodiments, higher sample volumes (e.g., greater than 250 microliters) may be collected and delivered with precision and accuracy.
Some embodiments may include electronic and/or embedded systems that may enable the automated sampling and processing system disclosed to interface automatically with third party test and/or sample systems, such as with third party microfluidics cartridges and testing systems, titer plates, and/or other third party systems. Some embodiments described herein may manage scheduling of future sampling events, may confirm the availability of aliquot sample targets (e.g., microfluidics cartridges) required to complete scheduled events, may confirm the aliquot sample targets are within their expiration dates per specifications provided by a corresponding manufacturer, may initiate various protocols to depyrogenate the pipette and process the sample, may discharge the used aliquot sample target, and may capture a test result from the third party test system.
Some embodiments of the online liquid autosampler and processing system described herein may allow for aliquot volumes from 250 microliters to 5 milliliters or more and may interface with various third party microfluidics systems, lab-on-a-chip systems, or support other filling and dispensing system needs.
Reference will now be made to the drawings to describe various aspects of some example embodiments of the invention. The drawings are diagrammatic and schematic representations of such example embodiments, and are not limiting of the present invention, nor are they necessarily drawn to scale.
The depyrogenation system 102 of
The pipette 104 of
The control/sense system 106 of
As an example, the control/sense system 106 may include a thermocouple or other temperature sensor in the depyrogenation system 102 and a timer. In this and other examples, the control/sense system 106 may be configured to operate the depyrogenation system 102 to heat a heater block of the depyrogenation system to a threshold temperature, such as at least 300° C., and to maintain the heater block at the threshold temperature for a threshold time, such as at least 30 minutes.
As another example, the control/sense system 106 may include one or more linear or rotary encoders or other position sensors configured to sense when the pipette 104 is in predetermined positions to aspirate a sample of the liquid and to dispense aliquots of the sample. In this and other examples, the control/sense system 106 may operate the pipette 104 to aspirate the sample and to dispense the aliquots when in the appropriate positions.
As another example, the control/sense system 106 may include one or more linear or rotary encoders or other position sensors configured to sense when the pipette 104 is in position to be rinsed and/or when the pipette 104 is clear of the sample source 108. In this and other examples, the control/sense system 106 may be configured to operate the rinse system 116 and/or the pipette 104 to rinse the pipette 104 and/or to operate the sample sanitizer 118 to sanitize the sample source 108 to reduce or eliminate formation of bacteria (e.g., bacterial film) on an exposed top surface of the liquid in the sample source 108.
The sample source 108 of
The movement system 110 may be communicatively coupled to the control/sense system 106 and/or may be controlled by the control/sense system 106. The movement system 110 may be coupled to one or more of the depyrogenation system 102, the pipette 104, the control/sense system 106, the sample source 108, the aliquot sample targets 112, the test system 114, the rinse system 116, and/or the sample sanitizer 118. The movement system 110 may be configured to position the pipette 104 within the depyrogenation system 102 for depyrogenation, position the tip of the pipette 104 within the sample source 108 to aspirate a sample of the liquid from the sample source 108, and/or position the pipette 104 to dispense an aliquot of the sample of the liquid into the aliquot sample targets 112. Alternatively or additionally, the movement system 110 may be configured to position the pipette 104 within the rinse system 116 to rinse the pipette 104 and/or to position the sample sanitizer 118 proximate the sample source 108 to sanitize the sample source. The movement system 110 may accomplish the positioning by moving any of the components of the autosampler 100 relative to the other. For instance, the pipette 104 may be positioned within the sample source 108 by moving the pipette 104 relative to the sample source 108, by moving the sample source 108 relative to the pipette 104, and/or by moving each relative to the other in at least one dimension (e.g., by moving the pipette 104 horizontally and by moving the sample source 108 vertically).
The movement system 110 may include one or more of a servo motor, a rotary actuator, a single axis actuator, a multi-axis actuator, or a robotic arm or any combination thereof configured to move one or more of the components of the autosampler 100. In an example implementation, for instance, the movement system 110 includes a single axis actuator mechanically coupled to the pipette 104 and a servo motor mechanically coupled to the sample source 108. The single axis actuator may be configured to selectively move the pipette 104 along a first axis (e.g., horizontally) to at least a first pipette position in which the pipette 104 is positioned within the depyrogenation system 102 and a second pipette position in which the pipette 104 is positioned in alignment with the sample source 108 along the first axis. In an example in which the sample source 108 includes a sample well with a funnel that includes a narrow bottom opening and a top opening, in the second pipette position the pipette 104 may be positioned in alignment with the funnel of the sample well along the first axis with the tip of the pipette 104 aligned along the first axis with the bottom opening of the funnel. The servo motor may be configured to selectively move the sample source 108 along a second axis (e.g., vertically) that is orthogonal to the first axis. In the example in which the sample source 108 includes the sample well, the servo motor may be configured to selectively move the sample well along the second axis when the pipette 104 is in the second pipette position to at least a first sample well position in which the tip of the pipette 104 is within the funnel above the bottom opening of the funnel and a second sample well position in which the tip of the pipette 104 is above the sample well.
The aliquot sample targets 112 may include microcartridges, titer plates, or other aliquot sample targets. As an example, some microcartridges that may be implemented as aliquot sample targets 112 may include four target wells or lanes where two of the four wells/lanes are positive controls and have endotoxin with chromogenic assay. Aliquots of the sample of the liquid may be dispensed into each of the four wells/lanes and then analyzed by the test system 114.
The test system 114 may generally be configured to analyze the aliquot sample targets 112, and more particularly the aliquots of the sample contained therein, for endotoxins. In some embodiments, the test system 114 may include a third party test device and/or system. As an example, the test system 114 may include a portable endotoxin testing system marketed by Charles River Laboratories, Inc. as the Endosafe®-PTS™ test system, or any other suitable test system. A description of the Endosafe®-PTS™ test system is described at http://www.criver.com/files/pdfs/emd/endotoxin/qc_en_d_endosafe_pts_datasheet.aspx (accessed on May 22, 2017), which is incorporated herein by reference in its entirety.
The rinse system 116 may be configured to rinse the pipette 104, including at least the tip of the pipette 104. The rinse system 116 may be configured to rinse the pipette 104 after each aspiration and dispense cycle. Some applications of endotoxin testing may, in the absence of the rinse system 116, result in carbonization of the tip of the pipette 104 due to small residuals of fluid being left at the end of the aspiration and dispense cycle. The residuals may be turned to carbon during the depyrogenation cycle. However, the rinse system 116 may be configured to rinse the pipette 104 after each aspiration and dispense cycle to remove the residuals that may otherwise lead to carbonization. In an example implementation, the rinse system 116 may include a pipette rinse well, a reservoir of cleansing fluid, a pump, and/or other suitable components. Optionally, the pipette rinse well may be coupled to and/or adjacent to the sample source 108 implemented as a sample well.
The sample sanitizer 118 may be configured to sanitize liquid in the sample source 108. The sample source 108 implemented as the sample well described in more detail below may be configured by itself to reduce and/or eliminate formation of bacteria (e.g., bacterial film) in or on the liquid in the sample source 108 in non-sterile environments. The sample sanitizer 118 may be implemented as a backup to further reduce and/or eliminate formation of bacteria (e.g., bacterial film) in or on the liquid in the sample source 108 in non-sterile environments.
The sample sanitizer 118 may include an ultraviolet (UV) sterilizing light emitting diode (LED) or other suitable sanitizer. The sample sanitizer 118 may be configured to be positioned proximate to the sample source 108 to illuminate the liquid in the sample source 108 with UV radiation or otherwise treat the liquid in the sample source 108. For instance, the sample sanitizer 118 may be configured to move between a first sanitizer position in which the sample sanitizer 118 is located over, and more particularly directly above, the sample source 108, and a second sanitizer position in which the sample sanitizer 118 is not located directly above the sample source 108. In some implementations, the sample sanitizer 118 may be moved to the second sanitizer position to allow the pipette 104 to aspirate a sample of the liquid and may be moved to the first sanitizer position between at least some aspirations.
Various example implementations of the autosampler 100 will now be described in turn.
As illustrated in
As illustrated in
The depyrogenation system 202 may include a depyrogenation chamber (not visible in
The control/sense system 206 may include some or all or any of the components previously described with respect to the control/sense system 106 of
The movement system 210 may include a single axis actuator 210A to move the pipette 204 along a first axis, e.g., horizontally, a servo motor 210B to move the sample well 208 along a second axis, e.g., vertically, and a microcartridge loader 210C to move microcartridges 217 from a microcartridge magazine 219 to an aliquot dispense position denoted at 221 in
The single axis actuator 210 may include a heat exchanger 202A coupled thereto. The heat exchanger 202A may be considered part of the depyrogenation system 202. The heat exchanger 202A may be configured to dissipate excess heat generated during the depyrogenation process.
The pipette 204 may be positionable by the movement system 210, and more particularly by the single axis actuator 210A, at various positions along the axis of the single axis actuator 210A. The axis of the single axis actuator 210A may be referred to as a first axis, an x axis, a horizontal axis, horizontal, or variations thereof. The various positions may include a first pipette position in which the pipette 204 is positioned within the depyrogenation system 202 and a second pipette position in which the pipette 204 is positioned in alignment with the sample well 208 along the first axis, e.g., horizontally. In the second pipette position, the tip of the pipette 204 may be aligned horizontally with a bottom opening of a funnel of the sample well 208. Alternatively or additionally, the various positions may include one or more third pipette positions in which the pipette 204 is positioned above, e.g., directly above, one or more target lanes of a corresponding one of the microcartridges 217 when loaded at the aliquot dispense position 221.
The sample well 208 may be positionable by the movement system 210, and more particularly by the servo motor 210B, at various positions along the axis of the servo motor 210B when the pipette 204 is in the second pipette position in alignment with the sample well 208. The axis of the servo motor 210B may be referred to as a second axis, a z axis, a vertical axis, vertical, or variations thereof. The various positions may include a first sample well position in which the tip of the pipette 204 is within the funnel of the sample well 208 above, e.g., directly above, the bottom opening of the funnel and a second sample well position in which the tip of the pipette 204 is above the sample well 208. In the second sample well position, the tip of the pipette 204 may directly above the bottom opening of the funnel of the sample well 208 but at a greater distance than in the first sample well position.
In the implementation of
The sample well 208 may additionally include an input (not shown in the view of
In operation, liquid to be sampled may be supplied by the input through the bottom opening 225 into the funnel 223. The liquid may generally flow upward and radially outward toward the output holes 229. The liquid may then exit the funnel 223 through the output holes 229 to be carried away by the output 231. Such a flow of the liquid through the sample well 208 may prevent and/or reduce the likelihood of contaminants being aspirated by the pipette 204. In particular, any contaminants (e.g., dust particles, etc.) that may inadvertently fall into or otherwise arrive in the liquid in the sample well 208 may be carried by the generally radially outward flow of liquid to the output holes 229 to exit the sample well 208.
As illustrated in
With reference to
The sample well 308 may include the same or similar funnel configuration with bottom opening in fluid communication with an input and one or more output holes around a perimeter at the top of the funnel in fluid communication with an output, similar to the sample well 208 discussed above. Alternatively or additionally, a pipette rinse well included as part of the rinse system 316 may be coupled and/or adjacent to the sample well 308, an example of which is discussed in more detail elsewhere herein with respect to another example online liquid autosampler (hereinafter “autosampler”) 400.
The movement system 310 may include a single axis actuator 310A, a servo motor 310B, a microcartridge loader 310C, and/or other components which may be the same or similar to components of the movement system 210 discussed elsewhere herein.
The testing system 314 may include a portable endotoxin testing system, such as the portable endotoxin testing system marketed by Charles River Laboratories, Inc. as the Endosafe®-PTS™ test system, or any other suitable test system.
The rinse system 316 may include the pipette rinse well (not visible in
As illustrated in
With reference to
Referring to
The heat exchanger 402A may be the same or similar to the heat exchanger 202A discussed elsewhere herein. For instance, the heat exchanger 402A may be coupled to a single axis actuator 410A of the movement system 410.
The heater block 402B may define a depyrogenation chamber 402E configured to accommodate therein the pipette 404, and more particularly, at least the tip of the pipette 404. The heater block 402B may include a copper heater block operated as a resistive heater. In other embodiments, other heaters operating on other principles may be implemented, such as inductive heating, convective heating, or other suitable heating technique.
The gate 402C may be movable between an open position (not shown) and a closed position as illustrated in
The insulation layer 402D may surround some or all of the heater block 402B and/or may be provided on an inside surface of the gate 402C. In some embodiments, the insulation layer 402D may include aerogel or other suitable insulator. The insulation layer 402D and/or the gate 402C may cooperate with the heater block 402B to enclose or at least substantially enclose the pipette 404, and more particularly at least the tip of the pipette 404, within the depyrogenation chamber 402E during depyrogenation.
Returning to
The pipette 404 may be positionable by the movement system 410, and more particularly by the single axis actuator 410A, at various positions along the axis of the single axis actuator 410A. The axis of the single axis actuator 410A may be referred to as a first axis, an x axis, a horizontal axis, horizontal, or variations thereof. The various positions may include a first pipette position in which the pipette 404 is positioned within the depyrogenation system 402 (e.g., within the depyrogenation chamber 402E of
The sample well 408 may be positionable by the movement system 410, and more particularly by the servo motor 410B, at various positions along the axis of the servo motor 410B when the pipette 404 is in the second pipette position in alignment with the sample well 408. The axis of the servo motor 410B may be referred to as a second axis, a z axis, a vertical axis, vertical, or variations thereof. The various positions may include a first sample well position in which the tip of the pipette 404 is within the funnel of the sample well 408 above, e.g., directly above, the bottom opening of the funnel and a second sample well position in which the tip of the pipette 404 is above the sample well 408. In the second sample well position, the tip of the pipette 404 may directly above the bottom opening of the funnel of the sample well 408 but at a greater distance than in the first sample well position.
In the implementation of
The sample well 408 may additionally include an input 430 and an output 431. The input 430 is also denoted in
In operation, liquid to be sampled may be supplied from the liquid supply tube 433 to the input 430 and then through the bottom opening 425 into the funnel 423. The liquid may generally flow upward and radially outward toward the output holes 429. The liquid may then exit the funnel 423 through the output holes 429 to be carried away by the output 431. Such a flow of the liquid through the sample well 408 may prevent and/or reduce the likelihood of contaminants being aspirated by the pipette 204. In particular, any contaminants (e.g., dust particles, etc.) that may inadvertently fall into or otherwise arrive in the liquid in the sample well 408 may be carried by the generally upward and radially outward flow of liquid to the output holes 429 to exit the sample well 408.
In some embodiments, in the second sample well position depicted in
The removal of excess liquid as described above may be referred to as a tip wipe protocol. The tip wipe protocol may alternatively or additionally be implemented after rinsing the tip of the pipette 404 in the pipette rinse well 416A. The pipette rinse well 416A may have an output coupled to the output 431 of the sample well 408 and an input coupled to a reservoir of cleansing liquid, such as the reservoir 316A of
The following experimental results are offered by way of illustration of the performance of an actual autosampler implemented as the autosampler 200 discussed in connection with
All bacterial endotoxin experiments were performed with an Endosafe®-PTS™ reader and Endosafe®-PTS™ cartridges (model PTS100, Charles River Laboratories, Inc., Wilmington, Mass., USA) and LAL reagent water (Lonza Biosciences, Walkersville, Md., USA). The Endosafe®-PTS™ reader was mounted onto the autosampler 200 and processing system and electronics interfaces were established. Ten Endosafe®-PTS™ cartridges were loaded into the cartridge magazine. Sampling events were scheduled and executed on the autosampler 200. Test results generated by the Endosafe®-PTS™ reader were captured and are displayed below.
For a valid assay the system must use certified, non-expired Endosafe®-PTS™ cartridges as defined by Charles River Laboratories, Inc. and be processed only on Endosafe®-PTS™ readers manufactured by Charles River Laboratories, Inc.
The reproducibility of sample collection and delivery was verified by delivered weight of sample product, examples of which are provided in Table 1 below.
Accuracy of delivery is dependent upon drop formation size which is dependent upon fluid viscosity and surface tension. The autosampler 200 utilizes the proximity of the delivery target (aliquot sample target 212) to the tip of the pipette 204 to facilitate and/or ensure that the target delivery volume is consistently removed. The proximity between the target and the tip is variable and can be adjusted to compensate for changes in fluid properties as well as target and pipette properties (materials, dimension changes).
The reproducibility of depyrogenation heating cycles was verified by thermocouple measurements within the pipette 404 while in the depyrogenation chamber.
The method 600 may be performed, in whole or in part, by the autosamplers 100, 200, 300, 400 and/or by other autosamplers. Alternatively or additionally, the method 600 may be implemented by a processor device that performs or controls performance of one or more of the operations of the method 600. For instance, a computer (such as the computing device 700 of
The method 600 may include one or more of blocks 602, 604, 606, 608, 610, and/or 612. Although illustrated as discrete blocks, various blocks may be divided into additional blocks, supplemented with additional blocks, combined into fewer blocks, or eliminated, depending on the particular implementation. The method 600 may begin at block 602.
In block 602 (“Position The Pipette Within The Depyrogenation System”), the pipette may be positioned within the depyrogenation system by the movement system. Block 602 may be followed by block 604.
In block 604 (“Depyrogenate The Pipette Within The Depyrogenation System At Least At A Threshold Temperature For At Least A Threshold Amount Of Time”), the pipette may be depyrogenated within the depyrogenation system at least at a threshold temperature for at least a threshold amount of time. The threshold temperature may be 250° C. and the threshold amount of time may be 30 minutes in an example embodiment. Block 604 may be followed by block 606.
In block 606 (“Position A Tip Of The Pipette Within The Sample Source”), a tip of the pipette may be positioned within the sample source using the movement system. Block 606 may be followed by block 608.
In block 608 (“Aspirating A Sample Of Liquid From Within The Sample Source”) a sample of liquid may be aspirated from within the sample source using the pipette. Block 608 may be followed by block 610.
In block 610 (“Positioning The Pipette Above An Aliquot Sample Target Using The Movement System”), the pipette may be positioned above an aliquot sample target using the movement system. Block 610 may be followed by block 612.
In block 612 (“Dispense An Aliquot Of The Sample Of The Liquid”), an aliquot of the sample of the liquid may be dispensed into the aliquot sample target.
One skilled in the art will appreciate that, for this and other processes, operations, and methods disclosed herein, the functions and/or operations performed may be implemented in differing order. Furthermore, the outlined functions and operations are only provided as examples, and some of the functions and operations may be optional, combined into fewer functions and operations, or expanded into additional functions and operations without detracting from the essence of the disclosed embodiments.
In some embodiments of the method 600 of
In these and other embodiments, the method 600 may further include providing a continuous flow of the liquid through the sample well. Providing a continuous flow of liquid through the sample well may include supplying the continuous flow of liquid from the input through the bottom opening of the funnel into the funnel; flowing the continuous flow of liquid through the funnel from the bottom opening of the funnel toward the top opening of the funnel to the one or more output holes; draining the continuous flow of liquid through the one or more output holes; and carrying away the drained continuous flow of liquid through the output. Alternatively or additionally, in these and other embodiments, positioning the tip of the pipette within the sample source using the movement system may include positioning the tip of the pipette within the funnel aligned with the bottom opening of the funnel and within a range of 10-18 mm above the bottom opening of the funnel.
Alternatively or additionally, the method 600 may further include dispensing the sample of the liquid from the pipette; positioning the tip of the pipette in a rinse well in fluid communication with a reservoir of cleansing liquid; and flowing the cleansing liquid through the rinse well to rinse the tip of the pipette. Alternatively or additionally, the method 600 may include performing the following sequence at least one time to further rinse the tip of the pipette: aspirating a portion of the cleansing liquid into the pipette while the tip of the pipette is positioned in the rinse well with the cleansing liquid flowing through the rinse well; positioning the tip of the pipette above the rinse well; and dispensing the aspirated portion of the cleansing liquid into the rinse well. Optionally, each sequence may include a time delay after dispensing the portion of the cleansing liquid before beginning the next sequence to ensure the cleansing liquid is able to be replenished in the pipette rinse well before the next aspiration of cleansing liquid. Alternatively or additionally, following the last rinse sequence, the tip wipe protocol may be implemented.
Alternatively or additionally, the method 600 may further include, after aspirating the sample of liquid from within the sample source and prior to positioning the pipette above the aliquot sample target, positioning the pipette above the sample source with a vertical distance in a range from 0.1 to 2 mm between an uppermost portion of the sample source and the tip of the pipette, where an excess portion of the liquid initially clings to an exterior of the tip of the pipette. The method 600 may further include moving the pipette past the upper portion of the sample source. The method 600 may further include removing, by the upper portion of the sample source, the excess portion of the liquid from the exterior of the tip of the pipette as the pipette moves past the upper portion of the sample source.
Alternatively or additionally, the method 600 may further include positioning a sample sanitizer over the sample well when the sample well is not in active use and sanitizing the sample well. For instance, a UV LED may be positioned over the sample well and may irradiate the sample well with UV radiation to sanitize the sample well.
Depending on the desired configuration, the processor 704 may be of any type including, such as a microprocessor (μP), a microcontroller (μC), a digital signal processor (DSP), or any combination thereof. The processor 704 may include one or more levels of caching, such as a level one cache 710 and a level two cache 712, a processor core 714, and registers 716. The processor core 714 may include an arithmetic logic unit (ALU), a floating point unit (FPU), a digital signal processing core (DSP Core), or any combination thereof. An example memory controller 718 may also be used with the processor 704, or in some implementations the memory controller 718 may be an internal part of the processor 704.
Depending on the desired configuration, the system memory 706 may be of any type, such as volatile memory (such as RAM), non-volatile memory (such as ROM, flash memory, or the like), or any combination thereof. The system memory 706 may include an operating system 720, one or more applications 722, and program data 724. The application 722 may include an autosampler algorithm 726 that is arranged to schedule and/or conduct autosampling operations associated with one or more of the autosamplers described herein. The program data 724 may include autosampler data 728 such as schedule autosampling events, threshold values, and/or other data that may be used to control aspects of the autosampling methods and/or operations described herein. In some embodiments, the application 722 may be arranged to operate with the program data 724 on the operating system 720 to perform one or more of the methods and/or operations described herein, including those described with respect to
The computing device 700 may include additional features or functionality, and additional interfaces to facilitate communications between the basic configuration 702 and any other devices and interfaces. For example, a bus/interface controller 730 may be used to facilitate communications between the basic configuration 702 and one or more data storage devices 732 via a storage interface bus 734. The data storage devices 732 may include removable storage devices 736, non-removable storage devices 738, or a combination thereof. Examples of removable storage and non-removable storage devices include magnetic disk devices such as flexible disk drives and hard-disk drives (HDDs), optical disk drives such as compact disk (CD) drives or digital versatile disk (DVD) drives, solid state drives (SSDs), and tape drives to name a few. Example computer storage media may include volatile and nonvolatile, removable and non-removable media implemented in any method or technology for storage of information, such as computer-readable instructions, data structures, program modules, or other data.
The system memory 706, the removable storage devices 736, and the non-removable storage devices 738 are examples of computer storage media. Computer storage media includes, but is not limited to, RAM, ROM, EEPROM, flash memory or other memory technology, CD-ROM, digital versatile disks (DVDs) or other optical storage, magnetic cassettes, magnetic tape, magnetic disk storage or other magnetic storage devices, or any other medium which may be used to store the desired information and which may be accessed by the computing device 700. Any such computer storage media may be part of the computing device 700.
The computing device 700 may also include an interface bus 740 for facilitating communication from various interface devices (e.g., output devices 742, peripheral interfaces 744, and communication devices 746) to the basic configuration 702 via the bus/interface controller 730. The output devices 742 include a graphics processing unit 748 and an audio processing unit 750, which may be configured to communicate to various external devices such as a display or speakers via one or more A/V ports 752. The peripheral interfaces 744 include a serial interface controller 754 or a parallel interface controller 756, which may be configured to communicate with external devices such as input devices (e.g., keyboard, mouse, pen, voice input device, touch input device, and/or others), sensors, or other peripheral devices (e.g., printer, scanner, and/or others) via one or more I/O ports 758. The communication devices 746 include a network controller 760, which may be arranged to facilitate communications with one or more other computing devices 762 over a network communication link via one or more communication ports 764.
The network communication link may be one example of a communication media. Communication media may typically be embodied by computer-readable instructions, data structures, program modules, or other data in a modulated data signal, such as a carrier wave or other transport mechanism, and may include any information delivery media. A “modulated data signal” may be a signal that includes one or more of its characteristics set or changed in such a manner as to encode information in the signal. By way of example, and not limitation, communication media may include wired media such as a wired network or direct-wired connection, and wireless media such as acoustic, radio frequency (RF), microwave, infrared (IR), and other wireless media. The term “computer-readable media” as used herein may include both storage media and communication media.
The computing device 700 may be implemented as a portion of a small-form factor portable (or mobile) electronic device such as a cell phone, a personal data assistant (PDA), a personal media player device, a wireless web-watch device, a personal headset device, an application-specific device, or a hybrid device that include any of the above functions. The computing device 700 may also be implemented as a personal computer including both laptop computer and non-laptop computer configurations.
The present disclosure is not to be limited in terms of the particular embodiments described herein, which are intended as illustrations of various aspects. Many modifications and variations can be made without departing from its spirit and scope, as will be apparent to those skilled in the art. Functionally equivalent methods and apparatuses within the scope of the disclosure, in addition to those enumerated herein, will be apparent to those skilled in the art from the foregoing descriptions. Such modifications and variations are intended to fall within the scope of the appended claims. The present disclosure is to be limited only by the terms of the appended claims, along with the full scope of equivalents to which such claims are entitled. It is to be understood that the present disclosure is not limited to particular methods, reagents, compounds, compositions, or biological systems, which can, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.
With respect to the use of substantially any plural and/or singular terms herein, those having skill in the art can translate from the plural to the singular and/or from the singular to the plural as is appropriate to the context and/or application. The various singular/plural permutations may be expressly set forth herein for sake of clarity.
The present invention may be embodied in other specific forms without departing from its spirit or essential characteristics. The described embodiments are to be considered in all respects only as illustrative and not restrictive. The scope of the invention is, therefore, indicated by the appended claims rather than by the foregoing description. All changes which come within the meaning and range of equivalency of the claims are to be embraced within their scope.
This application claims the benefit of and priority to U.S. Provisional App. No. 62/339,676, filed May 20, 2016, which is incorporated herein by reference.
| Number | Date | Country | |
|---|---|---|---|
| 62339676 | May 2016 | US |
