Sterile sampling method and device for automated cell engineering systems

Abstract

Devices and methods are provided for sterile sampling from automated cytotechnology systems. The sterile sampling device is configured to maintain the sterility of a sample reservoir during the intake and expulsion of fluids or other materials into the sterile sampling device. The methods provided herein utilize the sterile sampling device to achieve sterile withdrawal and injection of materials and fluids from and into the automated cytotechnology system.

Description

[Technical Field]

[0001] This disclosure Motion details In particular, the present disclosure relates to the use of cell engineering systems. Motion details Cell Engineering Systems and Other Nothing from the bacterial environment Nothing The present invention relates to methods and devices for providing microbial sampling. [Background technology]

[0002] As the clinical adoption of advanced cell therapies is expected to accelerate, more attention is being focused on the underlying manufacturing strategies that will enable these therapies to benefit patients worldwide.

[0003] The production of cells for cell therapy can require significant manual involvement due to the patient-specific nature of the product. In just one example, automating CAR T cell culture is particularly challenging due to the multiple delicate unit operations, including cell activation, transduction, and expansion.

[0004] Integrating cell activation, transduction, and expansion into a commercial manufacturing platform is critical to translating these important immunotherapies to broad patient populations. For these life-saving therapies to be applied to global patient populations, a transformation in manufacturing technology to support personalized medicine must be implemented. The benefits of automation have been previously documented. These benefits include not only labor-hour savings associated with the use of automation, but also improved product consistency, reduced room compartmentalization, a smaller cleanroom footprint, reduced complex training, and improved scale-up and logistics tracking.

[0005] The benefits of automation are realized when appropriate Nothing This application may not be fully realized without microbial process control. Motion details During operation related to cell engineering systems Nothing The solution provided herein provides a solution for maintaining the microbial activity of any type of Nothing During operation with the bacterial system Nothing It is more suitable for maintaining bacterial activity. Summary of the Invention

[0006] In an embodiment, Nothing A sterile plunger syringe is provided. Nothing The syringe plunger includes a syringe barrel defining a syringe reservoir and having an interconnection at a distal end and an opening surrounded by a syringe barrel flange at a proximal end; a syringe plunger including a syringe plunger rod flange, a plunger rod, and a reservoir face; a gasket disposed on the reservoir face and configured to provide a seal between the reservoir face and the syringe reservoir when the syringe plunger is seated within the syringe barrel; and a syringe plunger secured to the syringe barrel and configured to provide a syringe plunger seal. sealing and a device.

[0007] In a further embodiment, Motion details From cell engineering systems Nothing A method of microbial sampling is provided. The method includes a syringe barrel, a syringe plunger, and a syringe plunger seal. sealing Including devices Nothing providing a bacterial plunger syringe; Nothing Bacterial plunger syringe Motion details Connecting to the cell engineering system and Motion details Biosamples from cell engineering systems Nothing and withdrawing the syringe with a plunger.

[0008] In a further embodiment, Motion details From cell engineering systems Nothing A method of microbial sampling is provided, the method comprising a sample reservoir, a sample chamber, a filling device, and Nothing bacteria sealing Including the device, Nothing providing a bacterial sampling device; Nothing Bacteria sampling device Motion details It can be connected to a cell engineering system and automatically delivered via a filling device. Motion details and withdrawing the biological sample from the cytotechnology system. [Brief explanation of the drawings]

[0009] [Figure 1] 1 shows a generalized manufacturing process for cell culture. [Figure 2] 1 illustrates a laboratory space containing an exemplary cell engineering system, as described in embodiments herein. [Figure 3] 1 illustrates a cell culture production process that can be carried out within a cell engineering system, as described in embodiments herein. [Figure 4A] 1 shows an automated cell engineering system in a closed configuration. [Figure 4B] 1 shows a cassette that can be inserted into an automated cell engineering system. [Figure 4C] 1 shows an automated cell engineering system in an open configuration. [Figure 4D] 1 illustrates the location of cell culture chambers utilized within an automated cell engineering system. [Figure 4E] 1 illustrates the orientation of cell culture chambers utilized within an automated cell engineering system. [Figure 4F] 1 shows a more detailed view of a cell culture chamber utilized within an automated cell engineering system. [Figure 4G] 1 shows a process flow legend for an automated cell engineering system. [Figure 5A] 10 shows a disposable cassette that can be loaded into an automated cell engineering system in another configuration. [Figure 5B] 1 illustrates another configuration of the automated cell engineering system in an open configuration. [Figure 5C] 10 shows the cassette loaded into an automated cell engineering system in another configuration. [Figure 5D] 10 illustrates another configuration of the automated cell engineering system in a closed configuration. [Figure 5E] 10A-10C show detailed views of a cassette for use with an automated cell engineering system in another configuration. [Figure 6] 1 illustrates the use of a syringe and bag to sample from the cassette. [Figure 7] 1 illustrates a sterile sampling device consistent with embodiments herein. [Figure 8A] 1 illustrates a sterile sampling device consistent with embodiments herein. [Figure 8B] 1 illustrates a sterile sampling device consistent with embodiments herein. [Figure 9] 9A-9C illustrate additional sterile sampling devices consistent with embodiments herein. [Figure 10] 1 illustrates a further sterile sampling device consistent with embodiments herein. [Figure 11A] 1 illustrates a sterile sampling device consistent with embodiments herein. [Figure 11B] 1 illustrates a sterile sampling device consistent with embodiments herein. [Figure 11C] 1 illustrates a sterile sampling device consistent with embodiments herein. [Figure 11D] 1 illustrates a sterile sampling device consistent with embodiments herein. [Figure 11E] 1 illustrates a sterile sampling device consistent with embodiments herein. [Figure 12] FIG. 1 is a flow diagram illustrating a sterile sampling process consistent with embodiments herein. [Figure 13A] 1 illustrates an example of a sterile sampling process consistent with embodiments herein. [Figure 13B] 1 illustrates an example of a sterile sampling process consistent with embodiments herein. [Figure 13C] 1 illustrates an example of a sterile sampling process consistent with embodiments herein. [Figure 13D] 1 illustrates an example of a sterile sampling process consistent with embodiments herein. [Figure 13E] 1 illustrates an example of a sterile sampling process consistent with embodiments herein. [Figure 13F] 1 illustrates an example of a sterile sampling process consistent with embodiments herein. [Figure 13G] 1 illustrates an example of a sterile sampling process consistent with embodiments herein. [Figure 13H] 1 illustrates an example of a sterile sampling process consistent with embodiments herein. [Figure 13I] 1 illustrates an example of a sterile sampling process consistent with embodiments herein. DETAILED DESCRIPTION OF THE INVENTION

[0010] This disclosure Motion details In conjunction with the cell engineering system Nothing To provide a system and method for maintaining a microbial process. Motion details Cell engineering systems provide powerful tools for the production of a variety of modified cells and tissues, as well as biological materials (e.g., proteins, peptides, antibodies, antibody fragments, etc.). Motion details Among the benefits of cell engineering systems are: Nothing One example is the use of self-contained microbiological modules. Such modules provide a convenient and convenient way to process cells without the need to perform such processes in a high-level clean room. Nothing Creates a bacterial environment. Motion details Cell engineering systems are Nothing During operator interaction with these systems when operated in a sterile environment Nothing Steps must be taken to maintain bacterial integrity.

[0011] The methods and devices provided herein are Motion details From cell engineering systems Nothing facilitates bacterial sampling and therefore Motion details It allows for the removal and reinsertion of samples from the cell engineering system, such as cell cultures, biological material samples, reagent samples, and any other fluid or material samples.

[0012] One embodiment consistent with the present invention Motion detailsThe cell engineering system is the Cocoon™ platform, aspects of which are described in more detail below. The Cocoon™ platform is described in more detail in U.S. Patent Application No. 16 / 119,618, filed September 1, 2017, the contents of which are incorporated herein by reference in their entirety. Nothing Specific systems or systems that may be used with the bacterial sampling devices and methods Motion details The descriptions of cell engineering systems provided herein are merely exemplary. Nothing The microbial sampling devices and methods may be applied to additional systems, such as ADVA_X3® (Adva Biotechnology) and CLINIMACS PRODIGY® (Miltenyi Biotech). Nothing The bacterial sampling device and method are Nothing Any sample to be drawn from while maintaining bacterial integrity. Nothing Furthermore, the present invention can be suitably applied to bacterial systems. Nothing The bacterial sampling device and method are Nothing Any bacteria placed in a bacterial environment Nothing It can be suitably applied to bacterial systems.

[0013] As described herein, automated manufacturing facilities and comprehensive validation provide solutions to the logistical and operational challenges for the production of engineered cells and tissues. A key approach to introducing automation into a production process is to identify key modular steps, called "unit operations," where operators apply physical, biological, or chemical modifications to the production materials. In the case of cell manufacturing, this includes steps such as cell separation, genetic engineering, growth, washing, concentration, and cell harvest. Manufacturers often identify local process bottlenecks as immediate opportunities for introducing automation. This is reflected in the technical operating spectrum of most commercially available bioreactors, which tend to focus on individual process steps. Process Challenges in Cell Manufacturing ( Nothing The process of cellular chemistry (from maintaining sterility to tracking samples) is addressed herein with end-to-end automation that generates consistent cell output while ameliorating inevitable process variations. The methods described herein also provide simplification, and the associated electronic records aid in compliance with GMP standards.

[0014] Recent rapid progress in the clinical development of various cell cultures, including engineered autologous T cells for cancer immunotherapy, has led to the planning of related translational and scale-up / scale-out implications.

[0015] While specific cell culture growth protocols can vary for cell manufacturing, a generalized cell culture production process (including the production of autologous T cells) is shown in Figure 1. Figure 1 describes the unit operations of cell manufacturing, from the initial processing of, for example, a patient's blood sample to the formation of output cells for autologous T cell therapy.

[0016] To achieve cell manufacturing automation as described herein, sampling methods described herein are provided to understand the state of cells at each transition point and how they are affected by specific unit operations through the use of external analytical equipment. Microlot production for patient-specific therapies requires respecting key process sensitivities that affect the feasibility of automation. The automation described herein accommodates various process steps well.

[0017] A single, all-in-one system offers significant space efficiency gains and can minimize the required footprint within expensive GMP cleanrooms. For example, as shown in Figure 2, a fully integrated automated Verb The system is designed to maximize the required footprint and reduce expensive GMP cleanroom space. Figure 2, for example, shows 96 patient-specific end-to-end units running within a standard laboratory space.

[0018] As described herein, in embodiments, the methods provided utilize the COCOON platform (Octane Biotech, Kingston, ON), which integrates multiple unit operations in a single turnkey platform (see, e.g., U.S. Published Patent Application No. 2019 / 0169572, the disclosure of which is incorporated herein by reference in its entirety). However, it is understood that other fully or partially automated cell culture devices can be used in accordance with embodiments herein, including commercially available ones such as PRODIGY available from Miltenyi Biotech, Inc., XURI and SEFIA from General Electric Healthcare, and systems available from Atvio Biotech Ltd. Nothing The bacterial sampling device and method can be performed by each of the above and any other commercially available devices. Nothing It may be suitable for bacterial sampling operations.

[0019] The methods described herein can be used in conjunction with the production of CAR T cells (including activation, viral transduction and expansion, enrichment, and washing) in a fully integrated, closed, automated system (Figure 3).

[0020] In some embodiments, the methods described herein involve the use of functionally enclosed automated cells that suitably have instructions for performing the activation, transduction, expansion, enrichment, and harvesting steps of the cell culture. Motion details This is performed in conjunction with the cell engineering system 600 (see FIGS. 4A and 4B). Motion details Automated cell culture systems (also referred to as cell engineering systems) provide for the automated production of cell cultures. As used herein, "cell culture" refers to any suitable cell type, including individual cells, and multiple cells, or cells that can be formed into tissue structures. Exemplary cell cultures include blood cells, skin cells, muscle cells, bone cells, cells from various tissues and organs, and the like. In embodiments, genetically modified immune cells, including CAR T cells, can be produced as described herein. Exemplary autologous cell cultures include: Motion details The cell engineering system is also referred to throughout as COCOON or the COCOON system.

[0021] For example, a user can provide a cell engineering system pre-filled with cell culture materials and reagents (e.g., activation reagents, vectors, cell culture media, nutrients, and selection reagents, etc.), as well as parameters for cell production (e.g., starting number of cells, type of media, type of activation reagent, type of vector, and number or quantity of cells to be produced, etc.), and the cell engineering system can execute the method of producing engineered cell cultures, including genetically modified immune cell cultures, including CAR T cells, without further input from the user. At the end of the automated production process, the cell engineering system can alert the user (e.g., by playing a warning message or sending a mobile app alert) to collect the produced cells. In some embodiments, the functionally enclosed cell engineering system: NothingFunctionally enclosed refers to a system that is self-contained but may include means for gas exchange, e.g., hydrophobic filters and gas permeable tubing. In some embodiments, a functionally enclosed cytotechnological system is ... Nothing Minimizing contamination of cell cultures by reducing exposure to microbial environments. In additional embodiments, the functionally enclosed cell engineering system minimizes contamination of cell cultures by reducing user handling of the cells.

[0022] As described herein, the cell engineering system preferably includes a cassette 602 (see FIG. 4B). As used herein, "cassette" refers to a largely self-contained, removable, and replaceable element of the cell engineering system that includes one or more chambers for carrying out various elements of the methods described herein, and preferably also includes one or more of cell culture media, activation reagents, vectors, etc. The cassette may include a flexible bag, a rigid container, or other structural element. In some embodiments, the cassette may be configured to be disposable.

[0023] FIG. 4B illustrates an embodiment of a cassette 602 according to embodiments herein. In this embodiment, the cassette 602 includes a cold chamber 604 suitable for storing cell culture media and a hot chamber 606 suitable for performing activation, transduction, and / or expansion of immune cell cultures. Preferably, the hot chamber 606 is separated from the cold chamber 604 by a thermal barrier 1092 (see FIG. 5B). As used herein, "cold chamber" refers to a chamber maintained below room temperature, more preferably at about 2°C to about 8°C, to maintain cell culture media or the like at refrigerated temperatures. The cold chamber may include a medium bag or other holder containing about 1 L, about 2 L, about 3 L, about 4 L, or about 5 L of fluid. Additional medium bags or other fluid sources may be externally connected to the cassette via access ports, for example, by closed luer fittings, welded tubing, or the like.

[0024] As used herein, "high temperature chamber" refers to a chamber that is preferably maintained above room temperature, more preferably at a temperature that permits cell proliferation and growth, i.e., about 35-39°C, more preferably about 37°C.

[0025] In an embodiment, the high temperature chamber 606 suitably includes a cell culture chamber 610 (also referred to throughout as a proliferation chamber or cell growth chamber), as shown in Figures 4d and 4e.

[0026] The cassette, in some embodiments, may further include one or more fluidics pathways connected to the cell culture chambers, which fluidics pathways provide recirculation, waste removal, and uniform gas exchange and nutrient distribution to the cell culture chambers without disturbing the cells within the cell culture chambers. Cassette 602 also includes one or more pumps 605, including peristaltic pumps, for driving fluids through the cassette, and one or more valves 607 for controlling flow through the various fluidics pathways, as described herein.

[0027] In an exemplary embodiment, as shown in FIG. 4d, cell culture chamber 610 is a flat, non-flexible chamber (i.e., made of a substantially non-flexible material, such as plastic) that does not easily bend or flex. The use of a non-flexible chamber allows the cells to be maintained substantially undisturbed. As shown in FIG. 4e, cell culture chamber 610 is oriented to allow the cell culture to spread across the bottom 612 of the cell culture chamber. As shown in FIG. 4e, cell culture chamber 610 is preferably maintained in a position parallel to the floor or table, maintaining the cell culture undisturbed and allowing the cell culture to spread across a large area of ​​the bottom 612 of the cell culture chamber. In embodiments, the cell culture chamber may include features, such as a bow, to facilitate consistent filling and draining. In embodiments, the overall thickness of cell culture chamber 610 (i.e., chamber height 642) is low, on the order of about 0.5 cm to about 5 cm. As described herein, in exemplary embodiments, the cassette is pre-filled with one or more of cell culture media, media, activation reagents, and / or vectors (including any combination thereof). In further embodiments, these various elements can be added later via suitable injection ports or the like.

[0028] As described herein, in embodiments, the cassette preferably further includes one or more of a pH sensor, a glucose sensor, a dissolved oxygen sensor, a carbon dioxide sensor, a lactate sensor / monitor, and / or an optical density sensor. The cassette may also include one or more sampling and / or injection ports. Examples of such sampling and injection ports (1094) are shown in FIG. 5a and may include access ports for connecting the cartridge to external devices such as an electroporation unit or an additional media source. FIG. 5a also shows the location of a cell input 1095, a reagent warming bag 1096 that can be used to warm cell media, and an incubation zone 1107 that holds various components for use in the culture medium, including, for example, cell media, vectors, nutrients, and waste.

[0029] FIG. 5b shows the COCOON cytotechnology system with the cassette 602 removed. Components of the cytotechnology system are visible in FIG. 5b, including the gas control seal 1020, the warming zone 1021, the actuator 1022, the pivot 1023 for rocking or tilting the cytotechnology system as needed, and the cold zone 1024 for holding the cold chamber 604. Also shown is an exemplary user interface 1030, which may include readers for one-dimensional and two-dimensional codes, such as barcodes and QR codes, as well as the ability to use and receive input via a touchpad or other similar device. The user interface 1030 may further include a component identification sensor, such as a barcode reader, a QR code reader, a radio frequency ID interrogator, or other component identification sensor. In some embodiments, the cassette 602 may include a first identification component, such as a barcode, and the user interface 1030 may include a reader configured to read and identify the first identification component. FIG. 5e shows additional details of cassette 602, including the location of a secondary chamber 1150 that can be used if additional cell culture volume is needed, and a harvest chamber 1152 that can be used to harvest the final cell culture produced herein.

[0030] In an exemplary embodiment, as shown in FIG. 4f, the cell culture chamber 610 further comprises at least one of a distal port 620 configured to allow removal of air bubbles from the cell culture chamber and / or as a recirculation port, an intermediate port 622 configured to function as a recirculation inlet port, and a proximal port 624 configured to function as an exhaust port for cell removal.

[0031] In still further embodiments, the present disclosure provides a system including a cell culture chamber 610 for performing activation, transduction, and / or expansion of immune cell cultures, the cell culture chamber having a chamber volume configured to accommodate immune cell cultures, and a satellite volume 630 for increasing the working volume of the cell culture chamber by providing additional volume for medium and other working fluids without accommodating immune cell cultures (i.e., the satellite volume does not contain any cells). Motion details A cassette 602 for use in a cell engineering system 600 is provided.

[0032] FIG. 4g shows a schematic diagram illustrating the connections between the cell culture chamber 610 and the satellite volume 630. Also shown in FIG. 4g is the positioning of various sensors (e.g., pH sensor 650, dissolved oxygen sensor 651), as well as a sampling / sample port 652 and various valves (control valve 653, bypass check valve 654), as well as one or more fluidics pathways 640, preferably comprising silicone-based or other tubing components connecting the components. As described herein, the use of silicone-based tubing components allows oxygen delivery through the tubing components to facilitate gas transfer and optimal oxygenation for cell culture. Also shown in FIG. 4g is the use of one or more hydrophobic or hydrophilic filters 655 or 656 within the flow paths of the cassette, along with pump tubing 657 and a bag / valve module 658.

[0033] In an additional embodiment, as shown in Figure 4g, cassette 602 preferably further includes a cross-flow reservoir 632 for holding additional media, etc., as needed. Preferably, the cross-flow reservoir has a volume of about 0.50 ml to about 300 ml, more preferably about 100 ml to about 150 ml.

[0034] In some embodiments, the cell engineering system includes multiple chambers. In further embodiments, each of the activation, transduction, expansion, enrichment, and harvesting steps of the cell methods described herein is performed in a different one of the multiple chambers of the cell engineering system. In some embodiments, the cells are substantially undisturbed during transfer from one chamber to another. In other embodiments, the method steps are performed within the same chamber of the cell engineering system, and the cell engineering system automatically adjusts the chamber environment as needed for each step of the method, thus further enabling cells to remain undisturbed during the various steps.

[0035] Self Motion details The various processes and / or operations that take place in a cell engineering system are Motion details It may be necessary to withdraw material (such as cell cultures, biological samples, or other fluids) from the cell engineering system. Some processes and operations involve withdrawing such material automatically. Motion details It may be necessary to inject various materials back into the cell engineering system. Additional requirements include injecting transfection reagents and various materials into the system, such as nutrient supplements, media supplements, etc. Such processes and operations may be performed in a non- Nothing without introducing any contaminants from the bacterial environment Nothing configured to withdraw and return samples in a sterile fashion Nothing may benefit from a bacterial sampling device.

[0036] Figure 6 shows the Motion details 11 illustrates the use of a conventional syringe 1170 and bag 1172 being used to draw a sample from the cytology system 600. The conventional syringe Nothing bacteria of It may have an interior. Nothing The bacterial environment, i.e., the cell engineering system Nothing bacteria of than inside Nothing When operated in a low germ-free environment, the inside of the syringe NothingFor example, in conventional plunger syringes, dynamic seals are provided to protect the syringe interior over the range of motion of the syringe plunger. sealing However, a portion of the syringe interior behind the reservoir face is exposed to the environment during operation, i.e., when the syringe plunger is depressed. Contaminants clinging to the interior walls have the potential to be introduced into any material drawn into the syringe interior during plunger operation.

[0037] next, Nothing Specific embodiments of microbial sampling devices and methods are described with reference to Figures 7-13. Nothing With respect to the microbial sampling device, the terms "distal" and "proximal" are used in the following description in reference to a position or direction relative to the operator. Nothing "Proximal" and "proximally" refer to the input / output end of the microbial sampling device, whether away from or facing the operator under normal use. Nothing This position refers to the working end of the bacterial sampling device.

[0038] Embodiments herein include Nothing drawn into the bacterial sampling device and / or Nothing of the sample released from the bacterial sampling device Nothing Suitable for maintaining bacterial activity Nothing A bacterial sampling device is provided. Nothing The microbial sampling device includes a sample chamber defining a sample reservoir configured to contain a sample, such as a biological material sample, a reagent sample, a cell culture, or any other fluid. NothingThe microbial sampling device includes an interconnect at the distal end of the sample chamber that is configured to connect the sample chamber to other sampling equipment and may include, for example, weldable tubing, Luer-activated connectors such as ICU Medical Spiros and BD Q-Syte, genderless connectors such as CPC AseptiQuik, standard Luer lock fittings, twist fits, and any other suitable connectors.

[0039] Nothing The microbial sampling device further includes a filling device configured to draw a sample into the sample reservoir and / or expel a sample from the sample reservoir through manual or automated operation. A filling device consistent with embodiments herein may be automated. Motion details Filling devices include any device, mechanism, or system configured to provide a pressure differential (either overpressure or underpressure) between a sample reservoir connected to a reservoir of a cellular engineering system and the external environment. The pressure differential provided by the filling device drives the drawing and ejection of fluid from the sample reservoir. Filling devices are configured to connect to and operate with various sample chambers, as discussed herein. For example, as discussed below, a syringe sample chamber may utilize a syringe plunger as the filling device. Other sample chamber designs may utilize pumps, vacuums, syringes, and other mechanisms as the filling device, as discussed below.

[0040] Nothing The bacterial sampling device is Nothing During operation of the microbial sampling device, for example, during filling and emptying of the sample reservoir, the sample reservoir may be contaminated with environmental contaminants. Nothing Constructed to maintain bacterial activity Nothing bacteria sealing As used herein, the term "sample reservoir" refers to a device that includes a sample reservoir. Nothing Maintaining sterility prevents contamination of the sample reservoir from the environment and keeps the sample reservoir out of the environment. Nothing Higher than bacterial levels Nothing This refers to maintaining the bacteria level. Nothing bacteria sealing The device may or may not be in liquid contact.

[0041] Nothing Specific examples of various embodiments of the microbial sampling device are discussed in more detail below with respect to FIGS.

[0042] FIG. 7 is consistent with embodiments herein. Nothing The bacterial sampling device is shown in Figure 7. Nothing The bacterial sampling device is Nothing A syringe plunger 700 includes a syringe barrel 701, a syringe plunger 703, a gasket 705, and a syringe plunger sealing It may include a device 709.

[0043] Syringe barrel 701 is generally cylindrical and defines a syringe reservoir 702 that occupies the interior of syringe barrel 701. Syringe barrel 701 is an example of a sample chamber that defines a sample reservoir. The distal end of syringe barrel 701 includes interconnect 710. Interconnect 710 can be any type of syringe interconnect, including, for example, a luer lock tip, a slip tip, an eccentric tip, and a catheter tip. Any suitable syringe tip can be utilized as interconnect 710. The proximal end of syringe barrel 701 includes a proximal opening 711 surrounded by syringe barrel flange 707. Proximal opening 711 has substantially the same diameter as syringe reservoir 702. Syringe barrel 701 can include graduated volume markings to indicate the volume of material within syringe reservoir 702. Syringe barrel 701 may be made of any suitable material, including, for example, polyethylene, polycarbonate, polypropylene, stainless steel, or the like.

[0044] Syringe plunger 703 includes a syringe plunger rod 706, a syringe plunger rod flange 708 positioned at the proximal end of syringe plunger rod 706, and a reservoir face 704 positioned at the distal end of syringe plunger rod 706. Syringe plunger rod 706 may be manufactured from any suitable material, including, for example, polyethylene, polycarbonate, polypropylene, stainless steel, etc. Syringe plunger 703 is an example of a filling device configured to fill and empty syringe reservoir 702.

[0045] Gasket 705 is disposed on reservoir surface 704 and is configured to provide a seal between reservoir surface 704 and the interior wall of syringe barrel 701 when syringe plunger 703 is seated within syringe barrel 701. Thus, when syringe plunger 703 is seated within syringe barrel 701, gasket 705 seals syringe reservoir 702. Nothing When the plunger syringe 700 is actuated, the syringe plunger 703 is moved back and forth within the syringe reservoir 702 to take in and expel fluid, ie, liquid or gas.

[0046] The portion of the syringe reservoir 702 distal to the reservoir face 704 is referred to herein as the active portion of the syringe reservoir 702, while the portion of the syringe reservoir 702 proximal to the reservoir face 704 is referred to herein as the inactive portion of the syringe reservoir 702. Nothing The plunger syringe 700 is operated by expanding and contracting the volume of the active portion of the syringe reservoir 702 by moving the syringe plunger 703. The expansion and contraction of the volume of the active portion provides the pressure imbalance necessary to draw and expel fluid into the syringe reservoir 702.

[0047] Nothing The syringe plunger 700 includes a syringe plunger 712 secured to a syringe barrel 701 and configured to provide a syringe plunger seal 712. sealing The syringe plunger 709 is a syringe plunger used in the method of the present invention. sealing "Seal" in reference to a device refers to a fluid-tight, preferably liquid-tight, seal that in some embodiments does not allow the transfer of any substantial amount of fluid (including liquid and / or gas) through the seal, preferably to the extent of less than 1% of the volume of the reservoir 702, more preferably less than 0.1%, less than 0.01%, and even more preferably less than 0.001% of the volume of the reservoir 702. air Refers to a tight, joined, connected, or fitting. Syringe plunger sealing The device 709 is a syringe reservoir 702 Nothing Constructed to maintain bacterial activity Nothing bacteria sealing An example of a syringe plunger sealing The device 709 is Nothing The syringe plunger 703 surrounds the proximal end of the syringe 700. sealing The device 709 may be a tube, a bag, a balloon, a sock-like structure, an accordion, or the like. Shape pleated structure, or Nothing The plunger may be any other structure sufficient to enclose the proximal end of the syringe 700 and syringe plunger 703. sealing The device 709 may be constructed from any suitable material, including polymers, plastics, and the like.

[0048] Syringe plunger sealing The distal portion of the device 709 may be secured to the proximal portion of the syringe barrel 701 distal to the syringe barrel flange 707. sealing The device 709 may also be secured to the syringe barrel flange 707 itself. sealing The securing or attachment of device 709 to syringe barrel 701 may be achieved by adhesive, thermal bonding, chemical bonding, or sonic welding, by mechanical means such as clamping, and / or by any other suitable means. sealing The device 709 may be removably secured.

[0049] Syringe plunger sealing The proximal portion of the device 709 may optionally be fixed or attached to a syringe plunger rod flange 708. sealing Device 709 may entirely encompass or surround syringe plunger 703. In such an embodiment, syringe plunger rod flange 708 may be configured to allow movement of syringe plunger 703 to move the syringe plunger rod flange 708. sealing the syringe plunger to cause a corresponding movement of the device 709. sealing It may be connected to the device 709.

[0050] In a further embodiment, a syringe plunger sealing The device 709 has an opening at its proximal end. In such an embodiment, the syringe plunger sealing The opening at the proximal end of device 709 is sealed to syringe plunger rod flange 708 to maintain closure. The opening at the proximal end may be sealed to syringe plunger rod flange 708 with a syringe plunger rod flange seal (not shown).

[0051] In an embodiment, the syringe plunger seal 712 is substantially fluid-tight. Nothing In order to maintain bacterial activity, air In an embodiment, the syringe plunger seal 712 is substantially liquid-tight, but may be gas-tight, for example. S To enable bacteria, practically air In embodiments, the syringe plunger rod flange seal is substantially fluid-tight. In embodiments, the syringe plunger rod flange seal is substantially air In embodiments, the syringe plunger rod flange seal is substantially liquid-tight, but not substantially airPreferably, less than 1% of the volume of the reservoir 702 passes through the seal, more preferably less than 0.1%, less than 0.01%, and even more preferably less than 0.001% of the volume of the reservoir 702.

[0052] Syringe plunger sealing The device 709 is a syringe reservoir 702 Nothing Therefore, the syringe plunger sealing The device 709 is a syringe reservoir 702 Nothing Bacterial, packaged Nothing Early version of the 700 syringe plunger No Maintain or maintain the same level of bacterial activity, Nothing bacteria in In an environment without Nothing Bacteria may be caused by the operation of the plunger syringe 700 Nothing Prevents bacterial deterioration.

[0053] Conventional syringes operate by expanding and contracting the active portion of the syringe reservoir. As the active portion expands and contracts, the inactive portion of the syringe reservoir simultaneously contracts and expands, respectively. When the reservoir surface is retracted from the distal end to the proximal end of the syringe barrel, the inner wall of the syringe barrel, previously exposed to the environment outside the syringe, becomes the wall defining the active portion of the syringe reservoir. Therefore, contaminants from the environment outside the syringe can adhere to the wall of the syringe barrel and be introduced into the active portion of the syringe reservoir. Contaminants from the interior of the syringe can also adhere to the wall of the syringe barrel and thus escape into the environment. This can become more likely when the syringe plunger is repeatedly pushed in and out of the syringe barrel.

[0054] Syringe plunger sealing The device 709 is Nothing Isolating the inactive portion of the syringe reservoir 702 during operation of the sterile plunger syringe 700. By isolating the inactive portion of the syringe reservoir 702 from the surrounding environment, NothingContamination of syringe reservoir 702 from the environment in which sterile plunger syringe 700 is used can be prevented. Nothing The bacterial plunger syringe 700 is Nothing While the plunger syringe 700 is being operated, the syringe reservoir 702, both in the active and inactive portions, Nothing Maintains bacterial activity. Nothing The maintenance of sterility is achieved by interconnection 710 Nothing Sterile or sterile target It may need to be connected to the environment.

[0055] 8A-8B are further diagrams consistent with embodiments herein. Nothing 8A to 8B show a bacterial sampling device. Nothing The bacterial sampling device is Nothing A syringe plunger 800 includes a syringe barrel 801, a syringe plunger 803, a gasket 805, and a syringe plunger sealing It may include a device 809.

[0056] Syringe barrel 801 is generally similar to syringe barrel 701 and includes all of the same features. Syringe barrel 801 defines syringe reservoir 802 and includes interconnect 810, proximal opening 811 surrounded by syringe barrel flange 807. Syringe barrel 801 is an example of a sample chamber that defines a sample reservoir, and the syringe reservoir is an example of a sample reservoir.

[0057] Syringe plunger 803 is generally similar to syringe plunger 703 and includes all of the same features as syringe plunger 703, including syringe plunger rod 806, syringe plunger rod flange 808 positioned at the proximal end of syringe plunger rod 806, and reservoir face 804 positioned at the distal end of syringe plunger rod 806. Syringe plunger 803 is an example of a filling device configured to fill and empty syringe reservoir 802. Gasket 805 is disposed on reservoir face 804 and is generally similar to gasket 705. The portion of the syringe reservoir 802 distal to the reservoir surface 804 is referred to herein as the active portion 820 of the syringe reservoir 802 (shown in FIG. 8B), while the portion of the syringe reservoir 802 proximal to the reservoir surface 804 is referred to herein as the inactive portion 821 of the syringe reservoir 802 (shown in FIG. 8B). Nothing The plunger syringe 800 is operated by expanding and contracting the volume of the active portion 820 of the syringe reservoir 802 by moving the syringe plunger 803. The expansion and contraction of the volume of the active portion 820 provides the pressure imbalance necessary to draw and expel fluid into the syringe reservoir 802.

[0058] Nothing The syringe plunger 800 includes a syringe plunger 812 secured to a syringe barrel 801 and configured to provide a syringe plunger seal 812. sealing The syringe plunger further includes a device 809. sealing The device 809 is a syringe reservoir 802 Nothing Constructed to maintain bacterial activity Nothing bacteria sealing An example of a syringe plunger sealing Device 809 is Nothing The proximal end of the plunger syringe 800 and the syringe plunger 803 are enclosed.

[0059] Syringe plunger sealing Device 809 is a multi-accordion Shape Includes 813 pleats. Accordion Shape The pleats 813 are configured to stack when the syringe plunger 803 is advanced into the syringe reservoir 802, as shown in FIG. 8A, and to stretch when the syringe plunger 803 is withdrawn toward the proximal end of the syringe reservoir 802, as shown in FIG. 8B.

[0060] In an embodiment, a syringe plunger sealing The device 809 may be configured to have openings at both the proximal and distal ends. In such an embodiment, the syringe plunger sealing Device 809 is an accordion Shape The syringe plunger is a tube that is pleated by folds. sealing The distal end of device 809 is sealed to syringe barrel 801 with syringe plunger seal 812 on the proximal portion of syringe barrel 801. Syringe plunger seal 812 may be positioned on syringe barrel flange 807 or may be positioned on the cylindrical portion of syringe barrel 801, distal to syringe barrel flange 807. Syringe Plunger sealing The proximal end of the device 809 is sealed to the syringe plunger 803 with a syringe plunger rod flange seal 822. sealing The securing or attachment of device 809 to syringe barrel 801 and syringe plunger rod flange 808 may be achieved by adhesive, thermal bonding, chemical bonding, or sonic welding, by mechanical means such as clamping, and / or by any other suitable means. sealing The device 809 may be removably secured.

[0061] In an embodiment, a syringe plunger sealing The device 809 may be configured with a single opening at the distal end at the location of the syringe plunger seal 812. sealingThe closed proximal end of device 809 may fit over plunger rod flange 808 and may optionally be sealed to plunger rod flange 808 with syringe plunger rod flange seal 822. Syringe Plunger sealing The securing or attachment of device 809 to syringe barrel 801 and syringe plunger rod flange 808 may be achieved by adhesive, thermal bonding, chemical bonding, or sonic welding, by mechanical means such as clamping, and / or by any other suitable means. sealing The device 809 may be removably secured.

[0062] As described above with respect to syringe plunger seal 712, syringe plunger seal 812 and syringe plunger rod flange seal 822 are substantially fluid-tight, substantially air It may be liquid-tight or substantially liquid-tight, but air It doesn't have to be dense.

[0063] Syringe plunger sealing Device 809 is Nothing When the plunger syringe 800 is actuated, the syringe reservoir 802 Nothing It acts to maintain or preserve microbial activity. Nothing The maintenance of sterility is achieved by interconnection 810 Nothing Sterile or sterile target It may need to be connected to the environment.

[0064] In an embodiment, a syringe plunger sealing Device 809 is configured to enclose a substantially similar volume throughout the range of motion by syringe plunger 803. As used herein, substantially similar volume refers to a volume that varies by less than 10%, less than 5%, or less than 1%. When syringe plunger 803 is advanced into syringe reservoir 802 (as shown in FIG. 8A), syringe plunger sealingThe device 809 encloses an interior volume 830 (as shown in FIG. 8B), a portion of which is formed by the inactive portion 821 of the syringe reservoir 802, a portion of which is formed by the syringe plunger rod 806 and the syringe plunger sealing When syringe plunger rod 806 is withdrawn proximally through syringe reservoir 802 (as shown in FIG. 8B), the portion of interior volume 830 formed from inactive portion 821 is formed by the space and volume between syringe plunger rod 806 and syringe plunger rod 809. sealing The space between the syringe plunger rod 806 and the device 809 decreases in size while the volume formed by the space and volume increases. Throughout the range of motion of the syringe plunger rod 806, the internal volume 830 is maintained at substantially the same volume. As used herein, substantially the same volume means that any change in the volume of the internal volume 830 Nothing This means that the change in volume is not significant enough to interfere with the operation of the plunger syringe 800. For example, substantially the same volume may include a volume change of less than 5%, 4%, 3%, 2%, and / or 1%. If the volume of the internal volume 830 is different during operation, it may be Nothing This can create a pressure imbalance that prevents operation of the plunger syringe 800. Contracting the internal volume 830 is resisted by the pressure inside the internal volume 830, while expanding the internal volume 830 is resisted by the pressure outside the internal volume 830.

[0065] The internal volume 830 is the syringe plunger sealing Various design aspects of device 809 can maintain substantially the same volume throughout its range of motion. Shape The pleats 813 are sized and configured so that the interior volume 830 remains substantially the same size when collapsed and / or expanded. sealing The device 809 is a syringe plunger sealing The device 809 is flexible to allow the interior volume 830 to remain substantially the same volume during operation through flexing of the device 809. In a further embodiment, an accordion Shape Both methods may be utilized, with both pleats 813 and flexibility providing part of the maintenance of interior volume 830.

[0066] 9A-9C are consistent with embodiments herein. Nothing 1 shows a bacterial sampling device.

[0067] 9A to 9C are Nothing 9 shows an embodiment of a microbial sampling device 900. Nothing Bacterial sampling device 900 includes a sample container 901, a filter 930, and a filter interconnect 931. Figures 9A-9C show different container-filter connectors 950 / 951 / 952 and different filling devices 961 / 962 / 963. Nothing A bacterial sampling device 900 is shown.

[0068] Sample vessel 901 defines a sample reservoir 902 and includes an interconnect 910 and a proximal opening 911. Sample vessel 901 is an example of a sample chamber defining a sample reservoir. Sample vessel 901 may be a syringe reservoir or any other suitable vessel. Sample vessel 901 may be a syringe or other cylindrical vessel and may be constructed of any suitable material, including, for example, polyethylene, polycarbonate, polypropylene, stainless steel, etc. In embodiments, sample vessel 901 may include a flange 907, such as a syringe barrel flange. In embodiments, sample vessel 901 may include graduated volume markings.

[0069] The vessel filter connector 950 / 951 / 952 connects to the sample vessel 901 at the proximal opening 911. The vessel filter connector 950 / 951 / 952 provides a substantially fluid-tight seal, substantially air A hermetic seal, or a substantially liquid-tight seal air9B, receptacle filter connector 951 is a bayonet fitting that threads onto proximal opening 911 of sample vessel 901. In yet a further embodiment, receptacle filter connector 952 is integral with filter 930 and filter interconnect 931, as shown in FIG. 9C.

[0070] Receptacle filter connectors 950 / 951 / 952 connect to filter 930. In embodiments, filter 930 is a 0.22 micron hydrophobic filter. Other suitable filters may include a 0.2 micron hydrophobic filter, or any filter with a membrane hydrophobicity different from the fluid contained within the sampling device, where the membrane pore size is small enough to contain the fluid, but not large enough to accommodate the fluid. Nothing The filter 930 is large enough to allow air / gas to pass through the membrane so that the performance of the microbial sampling device is not substantially affected. Nothing Constructed to maintain bacterial activity Nothing bacteria sealing 1 is an example of a device. A filter interconnect 931 is disposed proximally of the filter 930.

[0071] Connected to filter interconnect 931 are filling devices 961 / 962 / 963 configured to fill and empty sample reservoir 902. Filling devices 961 / 962 / 963 may be another syringe, a pump, a syringe pump, a vacuum, or any other device suitable for drawing sample into a syringe reservoir through interconnect 910. For example, FIG. 9A illustrates the use of syringe filling device 961, FIG. 9B illustrates the use of pump filling device 962, and FIG. 9C illustrates the use of syringe pump filling device 963.

[0072] Nothing When the bacterial sampling device 900 is operated, the sample reservoir 902 Nothing Maintaining sterility. Maintaining sterility is achieved by ensuring that the interconnection 910 Nothing Sterile or sterile target The filter 930 may need to be connected to the environment. The filling device 961 / 962 / 963 is operated to reduce the pressure in the sample reservoir 902, causing the uptake of a sample or other fluid. To expel the sample or other fluid, the filling device 961 / 962 / 963 is operated to increase the pressure in the sample reservoir 902. The filter 930 allows sufficient gas (e.g., air) to pass through to provide the appropriate pressure reduction and increase, causing the uptake and expulsion of the sample or fluid. However, the filter 930 does not allow contaminants to pass through. Thus, the filter 930 Nothing During operation of the bacterial sampling device 900, the sample reservoir 902 Nothing The sample reservoir 902 operates to maintain sterility. Nothing The maintenance of sterility is achieved by ensuring that the filling device is Nothing bacteria to It does not need to be.

[0073] FIG. 10 is a diagram of a further embodiment consistent with the present disclosure. Nothing 1 shows a bacterial sampling device. Nothing Bacterial plunger syringe 1000 Nothing A bacterial sampling device. Nothing The plunger syringe 1000 includes a syringe barrel 1001 that defines a syringe reservoir 1002 and has an interconnect 1010 positioned at a distal end. The syringe barrel 1001 is an example of a sample chamber, and the syringe reservoir is an example of a sample reservoir.

[0074] Nothing Bacterial plunger syringe 1000 NothingIt further includes a sampling valve 1005 sealed to the syringe reservoir 1002 at its proximal opening 1011 by a bacterial seal 1012 . Nothing The bacterial seal 1012 is substantially fluid-tight, and in some embodiments, is substantially air In some embodiments, Nothing The bacterial seal 1012 is substantially liquid-tight, but air The sampling valve 1005 is operated to draw and expel fluid from the syringe reservoir 1002. Thus, the sampling valve 1005 is another example of a filling device configured to fill and empty the syringe reservoir 1002. Nothing The sampling valve 1005 in conjunction with the sterile seal 1012 is configured to prevent any contaminants from the environment from entering the syringe reservoir 1002. Thus, the sampling valve 1005 and Nothing The sterile seal 1012 is attached to the syringe reservoir 1002. Nothing Constructed to maintain bacterial activity Nothing bacteria sealing 1 is another example of a device.

[0075] During operation, the sampling valve 1005 is squeezed or compressed to expel air (or other gas) from the syringe reservoir 1002. When the sampling valve 1005 is released, it returns to its original shape, drawing fluid into the syringe reservoir 1002 through the interconnection 1010. Because the sampling valve 1005 and syringe reservoir 1002 are a closed system, contaminants cannot enter the syringe reservoir 1002 from the environment.

[0076] 11A-11E are diagrams illustrating further embodiments consistent with the present disclosure. Nothing 11A to 11E show a bacterial sampling device. Nothing The bacterial sampling device is Nothing A syringe plunger 1100 includes a syringe barrel 1101, a syringe plunger 1103, a gasket 1105, and a syringe plunger. sealing 11A and 11B show the syringe plunger 1103 fully inserted into the syringe barrel 1101. Nothing 11C and 11D are perspective and cutaway views of the syringe plunger 1100 with the syringe plunger 1103 fully withdrawn but still seated within the syringe barrel 1101. Nothing 11E is a perspective view and cutaway view of a syringe plunger 1100. sealing FIG. 11 is an enlarged view of device 1109.

[0077] Syringe barrel 1101 is generally similar to and includes all of the same features as syringe barrel 701 or syringe barrel 801. Syringe barrel 1101 defines syringe reservoir 1102 and includes proximal opening 1111 surrounded by interconnect 1110, syringe barrel flange 1107. Syringe barrel 1101 is an example of a sample chamber that defines a sample reservoir, and the syringe reservoir is an example of a sample reservoir.

[0078] Syringe plunger 1103 is generally similar to syringe plunger 703 and syringe plunger 803 and includes all of the same features as syringe plunger 703 and syringe plunger 803, including a syringe plunger rod 1106, a syringe plunger rod flange 1108 positioned at the proximal end of syringe plunger rod 1106, and a reservoir face 1104 positioned at the distal end of syringe plunger rod 1106. Syringe plunger 1103 is an example of a filling device configured to fill and empty syringe reservoir 1102. Gasket 1105 is disposed on reservoir face 1104 and is generally similar to gasket 705 and gasket 805. The portion of the syringe reservoir 1102 distal to the reservoir surface 1104 is referred to herein as the active portion 1120 of the syringe reservoir 1102 (shown in FIG. 11B), while the portion of the syringe reservoir 1102 proximal to the reservoir surface 1104 is referred to herein as the inactive portion 1121 of the syringe reservoir 1102 (shown in FIG. 11D). Nothing The plunger syringe 1100 is operated by expanding and contracting the volume of the active portion 1120 of the syringe reservoir 1102 by moving the syringe plunger 1103. The expansion and contraction of the volume of the active portion 1120 provides the pressure imbalance necessary to draw and expel fluid into the syringe reservoir 1102.

[0079] Nothing The syringe plunger 1100 is secured to a syringe barrel 1101 and configured to provide a syringe plunger seal 1112. sealing The syringe plunger further includes a device 1109. sealing The device 1109 is a syringe reservoir 1102 Nothing Constructed to maintain bacterial activity Nothing bacteria sealing An example of a syringe plunger sealing Device 1109 is Nothing A syringe plunger 1100 surrounds the proximal end of the syringe plunger 1103 .

[0080] Syringe plunger sealing The device 1109 is a flexible tube or bag sealed at a first end to the syringe barrel 1101 and at a second end to the syringe plunger rod flange. sealing Materials for constructing device 1109 include various plastics and polymers, preferably plastic tubes or bags having a thickness of about 100 mm to about 1-2 mm, e.g., about 100 mm to about 1 mm, or about 100 mm to about 800 mm.

[0081] Syringe plunger sealing The device 1109 is sealed to the syringe barrel 1101 with a syringe plunger seal 1112 on the proximal portion of the syringe barrel 1101. The syringe plunger seal 1112 may be located on the syringe barrel flange 1107 or may be located on the cylindrical portion of the syringe barrel 1101, distal to the syringe barrel flange 1107. Syringe Plunger sealing The proximal end of the device 1109 is sealed to the syringe plunger 1103 with a syringe plunger rod flange seal 1122. sealing The securing or attachment of the device 1109 to the syringe barrel 1101 and syringe plunger rod flange 1108 may be achieved by adhesive, thermal bonding, chemical bonding, or sonic welding, by mechanical means such as clamping, and / or by any other suitable means. sealing The device 1109 may be removably secured.

[0082] In an embodiment, a syringe plunger sealing The device 1109 may be configured to have openings at both the proximal and distal ends. In such an embodiment, the syringe plunger sealing The device 1109 is configured as a tube. In such an embodiment, a syringe plunger sealingThe ends of the device 1109 are closed by a syringe plunger seal 1112 and a syringe plunger rod flange seal 1122 .

[0083] In an embodiment, a syringe plunger sealing The device 1109 may be configured with a single opening at the distal end at the location of the syringe plunger seal 1112. sealing The closed proximal end of the device 1109 may fit over the plunger rod flange 1108 and may optionally be sealed to the plunger rod flange 1108 with a syringe plunger rod flange seal 1122. Syringe Plunger sealing The securing or attachment of the device 1109 to the syringe barrel 1101 and syringe plunger rod flange 1108 may be achieved by adhesive, thermal bonding, chemical bonding, or sonic welding, by mechanical means such as clamping, and / or by any other suitable means. sealing The device 1109 may be removably secured.

[0084] As described above with respect to syringe plunger seal 712, syringe plunger seal 1112 and syringe plunger rod flange seal 1122 are substantially fluid-tight, substantially air It may be liquid-tight or substantially liquid-tight, but air It doesn't have to be dense.

[0085] Syringe plunger sealing Device 1109 is Nothing When the plunger syringe 1100 is actuated, the syringe reservoir 1102 Nothing It acts to maintain or preserve microbial activity. Nothing The maintenance of sterility is achieved by interconnection 1110 Nothing Sterile or sterile target It may need to be connected to the environment.

[0086] In an embodiment, a syringe plunger sealingThe device 1109 is configured to enclose a substantially similar volume throughout the range of motion of the syringe plunger 1103. sealing The device 1109 encloses an interior volume 1130, the shape of which is Nothing The syringe plunger changes dynamically with the operation of the syringe plunger 1100. sealing The device 1109 is configured to lock the syringe plunger 1103 when the syringe plunger 1103 is withdrawn but still seated within the syringe barrel 1101 (as shown in FIGS. 11A and 11B). sealing The device 1109 is configured to have an inner diameter similar to the inner diameter of the syringe barrel 1101. In this configuration, the internal volume 1130 is sealing It is defined by the volume between the device 1109 and the syringe plunger rod 1106 plus any additional volume between the syringe plunger rod and the distal end of the syringe barrel 1101 .

[0087] When the syringe plunger 1103 is pushed into the syringe barrel 1101, the syringe plunger sealing Excess material from the device 1109 is forced into the syringe barrel 1101. Flexible syringe plunger sealing Device 1109 folds itself at pleats 1151. FIG. 11E shows pleats 1151 and the syringe plunger partially pushed into syringe barrel 1101. sealing Shown is a device 1109. Syringe plunger sealing The material of the device 1109 is thin and adheres to the wall of the syringe barrel 1101. Therefore, the syringe plunger within the syringe barrel 1101 sealing The portion of the device 1109 occupies only a negligible portion of the total volume inside the syringe barrel 1101. As used herein, negligible portion refers to a volume portion that is less than 5%, less than 2%, less than 1%, and / or less than 0.5% of the volume inside the syringe barrel 1101.

[0088] When the syringe plunger 1103 is partially depressed into the syringe barrel 1101, the internal volume 1130 is sealing The volume between the device 1109 and the portion of the syringe plunger rod 1106 that protrudes from the syringe barrel 1101, and the volume between the syringe barrel 1101 and the portion of the syringe plunger rod 1106 disposed within the syringe barrel 1101 (the collapsed syringe plunger sealing When the syringe plunger 1103 is fully depressed into the syringe barrel 1101, the internal volume 1130 is the volume between the syringe barrel 1101 and the portion of the syringe plunger rod 1106 disposed within the syringe barrel 1101 (excluding the collapsed syringe plunger sealing device 1109), and syringe plunger sealing It is defined by the remaining volume between the device 1109 and the portion of the syringe plunger rod 1106 that still protrudes from the syringe barrel 1101 .

[0089] In each of the above-described configurations, partially inserted, fully inserted, and fully withdrawn, and all configurations in between, the shape of the interior volume 1130 varies negligibly, so that the interior volume 1130 remains substantially the same. The length of the interior volume 1130, as represented by the distance between the back of the reservoir face 1104 and the syringe plunger rod flange 1108, does not change. The diameter of the interior volume 1130 is determined by the distance between the syringe barrel 1101 (the syringe plunger inside the syringe barrel 1101) in the various configurations. sealing 1109) and a syringe plunger similar in diameter to the syringe barrel 1101 as described above. sealingThe diameter of the internal volume 1130 varies negligibly because it is partially defined by the device 1109. As the syringe plunger 1103 moves in and out of the syringe barrel 1101, the relative proportion of the internal volume 1130 that is within the syringe barrel 1101 changes, but the approximate diameter of the internal volume 1130 does not change. Nothing The design of the plunger syringe 1100 remains substantially the same throughout any configuration.

[0090] In a further embodiment, a syringe plunger sealing The flexibility of device 1109 acts to balance the negligible volume change inside syringe barrel 1101 when syringe plunger 1103 is pushed into syringe barrel 1101. Syringe plunger occupies a portion of the interior of syringe barrel 1101. sealing Any reduction in the volume of the internal volume 1130 that may be caused by the device 1109 will result in the syringe plunger remaining outside the syringe barrel 1101. sealing A slight expansion of a portion of the device 1109 may be offset by a corresponding increase in the volume of the interior volume 1130, which may be caused, for example, through elongation or freedom of movement. sealing The flexibility of the device 1109 can help ensure that the internal volume 1130 remains substantially the same throughout all configurations of the syringe plunger 1103.

[0091] As mentioned above, the volume of the interior volume 1130 varies during operation, Nothing This can create a pressure imbalance that interferes with the operation of the plunger syringe 1100. Contracting the internal volume 1130 is resisted by the pressure inside the internal volume 1130, while expanding the internal volume 1130 is resisted by the pressure outside the internal volume 1130.

[0092] The various Nothing The bacterial sample device is a self-contained device as described herein. Motion detailsSuch a sample device is suitable for use in conjunction with cassette 602 of cell engineering system 600. Nothing bacteria of It allows for the removal and / or introduction of necessary or desired biological material samples, cells, media, etc. without risk of compromising the integrity and without disturbing the cells or biological material being produced therein.

[0093] In further embodiments, various of the methods described herein may be used. Nothing The bacterial sampling device Nothing While operating in a germ-free environment Nothing bacteria of It can be operated or utilized with any system that maintains the interior. Motion details The cell engineering systems discussed herein Nothing Bacterial concerns are functionally enclosed Nothing bacteria of This may equally apply to further systems, including internal Nothing The present invention may benefit from operation in a bacterial environment and, therefore, Nothing may benefit from the use of microbial sampling devices and methods.

[0094] In an embodiment, the Nothing Any or all of the microbial sampling devices may be operated manually or automatically, including via an operator's hand and / or using one or more tools, equipment, or devices configured to assist in operation. Nothing Automatic operation includes operating a microbial sampling device, as described herein. Nothing This includes the use of an actuator drive system or device to operate the microbial sampling device. Suitable actuators include syringe pumps, motors, servo motors, vacuum pumps, etc. Any or all of the above devices may be modified to accommodate automated operation. For example, for use with a syringe pump: NothingThe plunger syringe may include a mechanical portion configured to interface with an operational portion of a syringe pump. Nothing A bacterial sampling device may be provided. Nothing The bacterial sampling device includes an interconnect, a sample chamber and a sample reservoir, suitable fluid passages, suitable gas passages, suitable valves, a filling device configured to draw fluid into the sample chamber and expel fluid from the sample chamber, and an automated Nothing During operation of the bacterial sampling device Nothing Constructed to maintain bacterial activity Nothing bacteria sealing The apparatus may include a cartridge or cassette based sampling device.

[0095] Figure 12 shows Nothing FIG. 12 is a flow chart showing the steps of the bacterial sampling method. Nothing The bacterial sampling method 1200 may include any of the methods disclosed herein. Nothing This can be done manually or by automated methods using a bacterial sampling device. Nothing The microbial sampling method 1200 does not require that all of the operations discussed below be performed in the order described. Some operations may be performed in the order described. Nothing may be omitted, rearranged, and / or repeated multiple times without departing from the scope of the bacterial sampling method. Nothing The microbial sampling method 1200, as described herein, may be performed to withdraw a sample from a cellular engineering system. The withdrawn sample may include, for example, a cell culture, a biological material sample, a reagent sample, or any other fluid or medium that may need to be withdrawn from a cellular engineering system.

[0096] Nothing To perform the bacterial sampling method, Nothing A bacterial sampling device may be provided. Nothing Suitable for carrying out the bacterial sampling method 1200 NothingThe microbial sampling device defines a sample reservoir and includes an interconnect, a filling device, and Nothing fungal sealing The device may include at least a sample chamber. Nothing Examples of these aspects of the microbial sampling device are described herein and below. Nothing Suitable for carrying out the bacterial sampling method 1200.

[0097] In operation 1212, Nothing The bacterial sampling method 1200 is Nothing The method includes filling a sample reservoir of a microbial sampling device with a first gas portion, which may be air and / or any other suitable gas. The first gas portion comprises: Nothing bacteria in Also good and / or Nothing In an embodiment, the microorganism may be extracted from a bacterial environment. Nothing The bacterial sampling device is Nothing The sample reservoir may be packaged and provided to contain the first gas portion. In a further embodiment, a filling device may be utilized to fill the sample reservoir with the first gas portion.

[0098] In operation 1214, Nothing The bacterial sampling method 1200 is Nothing The method includes connecting a bacterial sampling device to a cytotechnology system. Nothing The interconnect of the bacterial sampling device is suitably connected to a port of the cassette of the cytotechnology system. Prior to connection, the port of the cytotechnology system is suitably cleaned, for example, with a sterile wipe or spray, to prevent contamination during the sampling procedure. Nothing bacteria to In an embodiment, Nothing The bacterial extension line Nothing It can be used to connect the interconnect of a bacterial sampling device to a port of a cytotechnology system. NothingConnections between the bacterial sampling device, extension line, and cytotechnology system port may be achieved via any suitable means, including luer locks, closure luer connectors, press fit connectors, snap fit connectors, and the like.

[0099] In operation 1216, Nothing The microbial sampling method 1200 includes injecting a sub-portion of the first gas portion through a filling device to fill a feed line in the cytotechnology system. Clear This includes: Nothing The microbial sampling device is operated to inject some or all of the first gas portion into the cell engineering system. The injected gas travels into the feed line of the cell engineering system and into the sample-containing system culture reservoir of the cell engineering system. The feed line of the cell engineering system is a conduit for the sample that connects the system culture reservoir to a port of the cell engineering system. Injecting gas through the feed line serves to push any sample collected in the feed line back into the system culture reservoir. This step can serve to homogenize the sample in the cell engineering system culture reservoir before sample withdrawal.

[0100] In operation 1218, Nothing The microbial sampling method 1200 includes, through operation of the filling device: Nothing mixing the sample in the cell engineering system with the bacterial sampling device. The sample in the cell engineering system culture reservoir is Nothing Further homogenization can be achieved through a mixing procedure applied through the bacterial sampling device. Nothing This can be accomplished by drawing into a bacterial sampling device and then injecting the mixed sample back into the cell engineering system culture reservoir. The drawing and re-injecting mixing can be performed any suitable number of times.

[0101] In operation 1220, Nothing The bacterial sampling method 1200 includes, via a filling device: NothingThe method includes drawing a sample from the cytotechnology system using a microbial sampling device. A specific amount of sample may be drawn. The drawing of the sample may be performed using a microbial sampling device. Clear The process may be carried out after the heating and / or mixing steps have been carried out.

[0102] In operation 1222, Nothing The microbial sampling method 1200 includes injecting a second portion of the gas into the cytotechnology system via a filling device, the second portion of the gas being injected into the cytotechnology system after drawing the sample to fill the feed line. Clear After drawing the sample through the cytotechnological system port, a portion of the sample may remain in the feed line. A second portion of the gas is Nothing The bacterial sampling device is injected back into the cytotechnological system and the feed line Clear possible.

[0103] As mentioned above, Nothing The microbial sampling method 1200 does not require that each of the operations 1212-1222 be performed in the order described. In various embodiments, any suitable arrangement of any number of operations 1212-1222 may be performed in the order described. Nothing For example, in one embodiment, Nothing The microbial sampling device automatically Motion details The sample can be connected to a cell engineering system and the sample can be transferred in operation 1222 without any intermediate operations. Nothing In a further embodiment, the bacteria may be extracted by a bacterial sampling device. Nothing The sample reservoir of the microbial sampling device may be filled with a first gas portion in operation 1212 and then automatically filled in operation 1214. Motion details A sub-portion of the first gas portion may then be automatically generated in operation 1220. Motion details Before drawing the sample from the cell engineering system, in operation 1216 Motion details After withdrawal, the second portion of the gas is automatically injected into the endoscopic system. Motion details Cell Engineering System Feedline Clear In order to do this, in operation 1222 Motion details In yet further embodiments, the injection of the second portion of gas may be omitted. Still further embodiments may include causing mixing in operation 1218 while excluding one or both of gas injection operations 1216 and 1222. In additional embodiments, the gas injection step is omitted, or Nothing All of the above embodiments may be implemented in operation 1212 either because the microbial sampling device is pre-filled with the first portion of gas. Nothing This may be performed without filling the microbial sampling device with the first portion of gas.

[0104] Nothing Bacteria sampling method 1200 and Nothing Throughout the operation of the bacterial sampling device, Nothing bacteria sealing The device is Nothing It is used to maintain the sterility of the sample reservoir. Nothing Maintaining sterility is essential for the integrity of any sample drawn into the sample reservoir. Nothing This entails maintaining the virulence of any sample contained within the cytotechnological system. Nothing Maintains bacterial activity. Nothing bacteria sealing Through the operation of the device, Nothing The bacterial sampling method 1200 is a method for detecting bacterial Nothing Without compromising the antibacterial properties Nothing It can be carried out in a bacterial environment.

[0105] 13A to 13K are the same as those described with respect to FIG. Nothing 13A-13K illustrate specific embodiments of bacterial sampling methods. The methods and procedures described with respect to Figures 13A-13K may also be performed through suitable automated means. As shown, Nothing The bacteria sampling device 1300 is Nothing Used in bacterial sampling procedures. NothingThe microbial sampling device 1300 is a Nothing The specific embodiment of Figures 13A-13K may include any of the microbial sampling devices, and any other devices capable of preforming the steps of the described procedure. Nothing 12 illustrates, but is not limited to, a bacterial sampling method 1200. As noted above, Nothing The microbial sampling method 1200 is not limited to the devices or actions shown or discussed with respect to Figures 13A-13K.

[0106] Figure 13A shows the Motion details External to port 1194 of cassette 602 of cell engineering system 600 Nothing 1 shows the first step of providing the bacteria. Nothing The bacterial sampling device is Nothing for sampling in bacterial environments Nothing A bacterial device is provided. Nothing Bacteria sampling device Motion details Before connecting to the cell engineering system, make sure port 1194 is Nothing bacteria to Being non Nothing Remove any contaminants from the bacterial environment.

[0107] Figure 13B shows Nothing Bacteria sampling device 1300 Motion details connecting to port 1094 of cassette 602 of cell engineering system 600; Motion details 10 illustrates the optional step of introducing or injecting a portion of gas into the cytotechnological system. Nothing Before connecting the bacterial sampling device 1300 to port 1094, some of the gas Nothing In an embodiment, the gas may be air or any other suitable gas. Nothing bacteria in For example, Motion details At least as strong as the environment of the cell engineering system Nothing bacteria in Part of the gas is Nothing Automatically from the bacterial sampling device 1300 Motion details Injected into the cell engineering system and feed line Clear Figure 13C shows the feed line 1310 containing fluid before air injection. The feed line 1310 contains fluid. Figure 13D shows the feed line 1310 after air injection with the fluid removed.

[0108] The feed line 1310 is connected to the gas injection to allow the proper mixture of fluids to be sampled. Clear For example, the fluid in the feed line 1310 may Motion details The fluid may not be adequately mixed with the fluid in the system culture reservoir of the cell engineering system 600. This may occur, for example, due to the addition (i.e., added fluid) or changes (e.g., cell growth) of fluid in the system culture reservoir. Therefore, flushing the feed line may result in a more homogenous fluid draw.

[0109] 13E and 13F are self- Motion details 13E illustrates the steps of mixing samples within the cell engineering system 600. As shown in FIG. 13E, the mixed sample is automatically Motion details In an embodiment, a specified volume is withdrawn from the cell engineering system 600 for a mixed sample. Nothing The microbial sampling device 1300 can be oriented so that the distal end having the interconnections faces upward. This orientation allows the drawn sample to Nothing It settles into the proximal portion of the sample reservoir of the microbial sampling device 1300, leaving the distal portion of the sample reservoir filled with gas.

[0110] Nothing Various orientations of the microbial sampling device 1300 are referenced with respect to the process shown in Figures 13A-13I. Nothing The orientation of the microbial sampling device may depend on the particular application. For example, NothingFluid entering the sampling reservoir from the top to seed the scaffolds within the bacterial sampling device may result in a more optimal distribution of cells on the scaffold. Alternatively, orienting the sampling device so that fluid enters the sampling reservoir from the bottom may provide less mixing to prevent shear stress on the fluid. Furthermore, the preferred orientation may depend on the specific design of the sampling device. For example, relying on a filter at one end Nothing For microbial sampling devices, it may be inappropriate to operate in an orientation that results in liquid contact with the filter. The orientations discussed herein are suitable for the operations and devices discussed herein, but may be varied according to alternative requirements without departing from the scope of the present disclosure.

[0111] After the sample is extracted, Nothing Use the Bacterial Sampling Device 1300 to automatically collect mixed samples. Motion details and then injected back into the cell engineering system 600. Motion details 6 shows a mixed sample being injected back into the cell engineering system 600. In an embodiment, Nothing The bacterial sampling device 1300 is oriented so that the distal end with the interconnection faces downward to inject the mixed sample. This orientation allows the withdrawn sample to settle in the distal portion of the sample reservoir, leaving the proximal portion of the sample reservoir filled with gas. Motion details When pushed back into the cell engineering system, Nothing The remaining air in the bacterial sampling device 1300 continues to sample and enters the feed line 1310. Clear The steps of withdrawing and injecting may optionally be repeated multiple times.

[0112] The withdrawal and injection steps are performed automatically. Motion details The cassette 602 of the cell engineering system 600 acts to induce mixing within the system culture reservoir. Motion details Mixing fluids within the system culture reservoirs of cassette 602 of cell engineering system 600 may result in a more homogenous sample draw. Motion details The samples in the system culture reservoirs of the cassette 602 of the cell engineering system 600 are automatically Motion details via mechanical means contained within cassette 602 of cell engineering system 600, and any other suitable method. Motion details Mixing can be achieved in alternative ways, such as by rocking or shaking the cassette 602 of the cell engineering system 600 or the system culture reservoir.

[0113] Figure 13G shows the Motion details 6 illustrates the step of withdrawing a sample from cassette 602 of cell engineering system 600. A specific volume of sample is withdrawn. In an embodiment, Nothing The microbial sampling device 1300 can be oriented so that the distal end having the interconnections faces upward. This orientation allows the drawn sample to Nothing It settles into the proximal portion of the sample reservoir of the microbial sampling device 1300, leaving the distal portion of the sample reservoir filled with gas.

[0114] This orientation may enhance the ability to draw a specific sample volume. As sample is drawn through the extension tubing and interconnect, it falls into the sample reservoir, Nothing The volume can be easily verified by comparing it with the graduated markings on the bacterial sampling device.

[0115] Figure 13H shows the automatic extraction of part of the gas after the sample was extracted. Motion details 6 shows the step of injecting the sample into the cassette 602 of the cell engineering system 600. After the sample is extracted, Nothing The microbial sampling device 1300 can be oriented so that the interconnect faces upward. This orientation causes the sample to fall into the proximal end of the sample reservoir, leaving the distal portion of the sample reservoir filled with gas. The gas then self-replenishes. Motion details The cassette 602 of the cell engineering system 600 is pushed back into the feed line 1310 as shown in FIG. 13H. Clear do.

[0116] Figure 13I shows the sample transfer step. After withdrawing the appropriate amount of sample, Nothing The bacteria sampling device 1300 is Motion details The sample may be cut from the cell engineering system 600. Nothing The sample may be dispensed from the microbial sampling device 1300 into a suitable sample container for storage, shipping, analysis, or any further processing. In embodiments, the sample may be dispensed into a suitable container for storage prior to shipping, analysis, or further processing. Nothing It may remain within the bacterial sampling device 1300.

[0117] As described with respect to method 1200 and the illustrated steps shown in FIGS. 13A-13I, consistent with embodiments herein. Nothing The bacterial sampling method is Motion details Into and by the cell engineering system 600 Motion details This may involve multiple sample transfers from the cell engineering system 600. Nothing Any contaminants introduced into the bacterial sampling device will Motion details Such contamination can be introduced into the cell engineering system 600. Motion details This may damage or destroy the cell cultures or other materials being processed in the cell engineering system 600. Nothing bacteria sealing The devices discussed herein include Nothing The bacterial sampling device must be kept in the same position throughout the sampling process. Nothing It is used to maintain bacterial activity. Nothing Sample reservoir of the bacterial sampling device Nothing The maintenance of fungi Nothing Contamination of the sample reservoir is prevented when the microbial sampling device is operated to take in and expel a sample. Nothing Provided by the bacterial sampling device Nothing The maintenance of bacterial activity is Motion details Whether it is a mixed sample intended to be returned to the cell engineering system 600 or a sample intended for later processing, Nothingof any sample drawn into the bacterial sampling device Nothing Therefore, the present invention provides a method for the preparation of a medicament for the prevention of bacterial infection. Nothing The maintenance of bacterial activity is Motion details of the cell engineering system 600 and any samples withdrawn for subsequent processing. Nothing Helps maintain bacterial integrity.

[0118] Illustrative Embodiments Additional embodiments include the following:

[0119] Embodiment 1 includes a syringe barrel defining a syringe reservoir and having an interconnection at a distal end and an opening surrounded by a syringe barrel flange at a proximal end; a syringe plunger including a syringe plunger rod flange, a plunger rod, and a reservoir face; a gasket disposed on the reservoir face and configured to provide a seal between the reservoir face and the syringe reservoir when the syringe plunger is seated within the syringe barrel; and a syringe plunger secured to the syringe barrel and configured to provide a syringe plunger seal. sealing a device; Nothing It is a bacterial plunger syringe.

[0120] Embodiment 2 is a syringe plunger sealing 2. The method of claim 1, wherein the device is secured to a proximal portion of the syringe barrel. Nothing It is a bacterial plunger syringe.

[0121] Embodiment 3 is a syringe plunger sealing 2. The method of claim 1, wherein the device is secured to a syringe barrel flange. Nothing It is a bacterial plunger syringe.

[0122] Embodiment 4 is a syringe plunger sealing 4. The method of claim 3, wherein the device is secured by adhesive, thermal bonding, chemical bonding, or sonic welding. Nothing It is a bacterial plunger syringe.

[0123] Embodiment 5 is a syringe plunger sealing 4. The method of claim 1, wherein the device is removably secured by clamping. Nothing It is a bacterial plunger syringe.

[0124] Embodiment 6 is a syringe plunger sealing The device has multiple accordions Shape 6. The method according to any one of claims 1 to 5, comprising pleating. Nothing It is a bacterial plunger syringe.

[0125] Embodiment 7 is a syringe plunger sealing 7. The method of claim 1, wherein the device seals the opening at the proximal end of the syringe reservoir. Nothing It is a bacterial plunger syringe.

[0126] Embodiment 8 is the syringe according to any one of embodiments 1 to 7, wherein the syringe plunger seal is substantially fluid-tight. Nothing It is a bacterial plunger syringe.

[0127] Embodiment 9 is a method for manufacturing a syringe plunger seal, the method comprising: air 9. The method of claim 8, wherein the Nothing It is a bacterial plunger syringe.

[0128] Embodiment 10 is a self- Motion details From cell engineering systems Nothing A method of microbial sampling, the method comprising: a syringe barrel; a syringe plunger; and a syringe plunger seal. sealing Including devices Nothing providing a bacterial plunger syringe; Nothing Bacterial plunger syringe Motion details Connecting to the cell engineering system and Motion details Biosamples from cell engineering systems Nothing and withdrawing the syringe plunger.

[0129] Embodiment 11 is a syringe barrel defining a syringe reservoir having an interconnection at a distal end and an opening surrounded by a syringe barrel flange at a proximal end, wherein the syringe plunger comprises a syringe plunger rod flange, a plunger rod, a reservoir face, a gasket disposed on the reservoir face and configured to provide a seal between the reservoir face and the syringe reservoir when the syringe plunger is seated in the syringe barrel, and a gasket secured to the syringe barrel and configured to provide a syringe plunger seal. sealing 11. The method of embodiment 10, comprising a device.

[0130] Embodiment 12 is a syringe plunger sealing 12. The method of embodiment 11, wherein the device seals the opening at the proximal end of the syringe reservoir.

[0131] Embodiment 13 is Nothing Bacterial plunger syringe Motion details Before connecting to the cell engineering system, some of the gas Nothing 13. The method of any one of embodiments 10 to 12, further comprising introducing the syringe plunger into a syringe.

[0132] Embodiment 14 is a syringe plunger seal, the syringe plunger seal is an internal Nothing 14. The method according to any one of embodiments 10 to 13, wherein the method maintains bacterial activity.

[0133] Embodiment 15 is a syringe reservoir internal Nothing Maintaining bacterial Nothing 15. The method of any one of embodiments 14, wherein contamination of the syringe reservoir is prevented when the sterile plunger syringe is actuated.

[0134] In the sixteenth embodiment, after the biological sample is extracted, a portion of the gas is injected to automatically Motion details Feed lines in cell engineering systems Clear16. The method of any one of embodiments 10 to 15, further comprising:

[0135] Embodiment 17 injects a portion of the gas before drawing the biological sample to automatically Motion details Feed lines in cell engineering systems Clear 17. The method of any one of embodiments 10 to 16, further comprising:

[0136] Embodiment 18 further comprises, before drawing the biological sample, Nothing Pump the syringe plunger to Motion details 18. The method of any one of embodiments 10 to 17, further comprising mixing in a cell engineering system.

[0137] Embodiment 19 is Nothing Pumping the syringe plunger automatically mixes the sample. Motion details From cell engineering systems Nothing The syringe plunger automatically draws the mixed sample into the syringe. Motion details 18. The method of embodiment 17, comprising returning the cell to the cell engineering system.

[0138] Embodiment 20 is a self- Motion details From cell engineering systems Nothing 1. A method of microbial sampling, the method comprising: a sample chamber defining a sample reservoir; a filling device; and Nothing bacteria sealing Including the device, Nothing providing a bacterial sampling device; Nothing Bacteria sampling device Motion details It can be connected to the cassette of the cell engineering system and automatically inserted via the filling device. Motion details and withdrawing a biological sample from the cytotechnology system.

[0139] Embodiment 21 is Nothing Before connecting the microbial sampling device to the cassette, a portion of the gas is Nothing21. The method of embodiment 20, further comprising introducing into a microbial sampling device.

[0140] Embodiment 22 is Nothing bacteria sealing While drawing a biological sample through the device, the inside of the sample reservoir Nothing 22. The method of embodiment 20 or 21, further comprising maintaining microbial activity.

[0141] Embodiment 23 is a sample reservoir internal Nothing Maintaining bacterial Nothing 23. The method of embodiment 22, wherein contamination of the sample reservoir is prevented when the microbial sampling device is operated.

[0142] In the twenty-fourth embodiment, after the biological sample is extracted, a portion of the gas is injected to automatically Motion details Feed lines in cell engineering systems Clear 24. The method of any one of embodiments 20 to 23, further comprising:

[0143] Embodiment 25 injects a portion of the gas before drawing the biological sample to automatically Motion details Feed lines in cell engineering systems Clear 25. The method of any one of embodiments 20 to 24, further comprising:

[0144] Embodiment 26 is a method for automatically extracting a biological sample before drawing the biological sample through a filling device. Motion details 26. The method of any one of embodiments 20 to 25, further comprising mixing in a cell engineering system.

[0145] Embodiment 27 is a method for mixing a liquid containing a mixture of a plurality of liquids. Motion details Drawing the mixed sample from the cell engineering system into a sample reservoir and automatically discharging the mixed sample. Motion details 27. The method of embodiment 26, wherein the method is induced by returning the cell to an engineered system.

[0146] It will be readily apparent to those skilled in the relevant art that other suitable modifications and adaptations to the methods and applications described herein can be made without departing from the scope of any of the embodiments.

[0147] Although particular embodiments have been illustrated and described herein, it is to be understood that the claims should not be limited to the specific forms or arrangements of parts described and illustrated. Although exemplary embodiments are disclosed and specific terms are employed herein, they are used in a generic and descriptive sense only and not for purposes of limitation. Modifications and variations of the embodiments are possible in light of the above teachings. It is therefore to be understood that the embodiments may be practiced otherwise than as specifically described.

[0148] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference.

Claims

1. A sterile plunger syringe, a syringe barrel defining a syringe reservoir and having an interconnect at a distal end and an opening surrounded by a syringe barrel flange at a proximal end; a syringe plunger including a syringe plunger rod flange, a plunger rod, and a reservoir face; a gasket disposed on the reservoir surface and configured to provide a seal between the reservoir surface and the syringe reservoir when the syringe plunger is seated within the syringe barrel; a syringe plunger sealing device secured to the syringe barrel and configured to provide a syringe plunger seal; A sterile plunger syringe, wherein the syringe plunger sealing device is configured to enclose an interior volume that remains substantially the same throughout a range of motion of the syringe plunger.

2. 10. The sterile plunger syringe of claim 1, wherein the syringe plunger sealing device is secured to a proximal portion of the syringe barrel.

3. 10. The sterile plunger syringe of claim 1, wherein the syringe plunger sealing device is secured to the syringe barrel flange.

4. 4. The sterile plunger syringe of claim 1, wherein the syringe plunger sealing device is secured by adhesive, thermal bonding, chemical bonding, or sonic welding.

5. 4. A sterile plunger syringe according to any one of claims 1 to 3, wherein the syringe plunger sealing device is removably secured by clamping.

6. 6. The sterile plunger syringe of claim 1, wherein the syringe plunger sealing device comprises a plurality of accordion-like pleats.

7. 7. The sterile plunger syringe of claim 1, wherein the syringe plunger sealing device seals the opening at the proximal end of the syringe reservoir.

8. A sterile plunger syringe according to any one of claims 1 to 7, wherein the syringe plunger seal is substantially fluid-tight.

9. 9. The sterile plunger syringe of claim 8, wherein the syringe plunger seal is substantially airtight.

10. 1. A method of aseptic sampling from an automated cell engineering system, said method comprising: Providing a sterile plunger syringe according to claim 1; connecting the sterile plunger syringe to the automated cell engineering system; and withdrawing a biological sample from the automated cell engineering system with the sterile plunger syringe.

11. 11. The method of claim 10, wherein the syringe barrel defines a syringe reservoir having an interconnection at a distal end and an opening surrounded by a syringe barrel flange at a proximal end, the syringe plunger includes a syringe plunger rod flange, a plunger rod, a reservoir face, and a gasket disposed on the reservoir face and configured to provide a seal between the reservoir face and the syringe reservoir when the syringe plunger is seated within the syringe barrel, and the syringe plunger sealing device is secured to the syringe barrel and configured to provide the syringe plunger seal.

12. The method of claim 11 , wherein the syringe plunger sealing device seals the opening at the proximal end of the syringe reservoir.

13. 13. The method of any one of claims 10 to 12, further comprising introducing a portion of sterile gas into the sterile plunger syringe before connecting the sterile plunger syringe to the automated cell engineering system.

14. The method of any one of claims 10 to 13, wherein the syringe plunger seal maintains sterility of the interior of the syringe reservoir.

15. 15. The method of claim 14, wherein maintaining sterility of the interior of the syringe reservoir prevents contamination of the syringe reservoir when the sterile plunger syringe is actuated.

16. 16. The method of any one of claims 10 to 15, further comprising injecting a portion of gas to clear feed lines in the automated cell engineering system after withdrawing the biological sample.

17. 17. The method of any one of claims 10 to 16, further comprising injecting a portion of gas to clear feed lines in the automated cell engineering system before withdrawing the biological sample.

18. 18. The method of any one of claims 10 to 17, further comprising pumping the sterile plunger syringe to cause mixing within the automated cell engineering system prior to withdrawing the biological sample.

19. Pumping the sterile plunger syringe comprises: withdrawing a mixed sample from the automated cytotechnology system into the sterile plunger syringe; and returning the mixed sample to the automated cytotechnology system.

20. 1. A method of aseptic sampling from an automated cell engineering system, said method comprising: Providing a sterile plunger syringe according to claim 1; connecting the sterile plunger syringe to a cassette of the automated cell engineering system; and withdrawing a biological sample from the automated cell engineering system via the syringe plunger.

21. 21. The method of claim 20, further comprising introducing a portion of gas into the sterile plunger syringe before connecting the sterile plunger syringe to the cassette.

22. 22. The method of claim 20 or 21, further comprising maintaining sterility of the interior of the syringe reservoir while withdrawing the biological sample through the syringe plunger sealing device.

23. 23. The method of claim 22, wherein maintaining sterility of the interior of the syringe reservoir prevents contamination of the syringe reservoir when the sterile plunger syringe is actuated.

24. 24. The method of any one of claims 20 to 23, further comprising injecting a portion of gas to clear feed lines in the automated cell engineering system after withdrawing the biological sample.

25. 25. The method of any one of claims 20 to 24, further comprising injecting a portion of gas to clear feed lines in the automated cell engineering system before withdrawing the biological sample.

26. 26. The method of any one of claims 20 to 25, further comprising causing mixing within the automated cell engineering system prior to withdrawing the biological sample via the syringe plunger.

27. The mixing drawing a mixed sample from the automated cell engineering system into the syringe reservoir; and returning the mixed sample to the automated cytotechnology system.

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