Bioreactor well access

Abstract

A system for preparing microbial cultures includes a microtiter plate assembly including a plurality of openings, and a plurality of plugs sized and configured to be received within the plurality of openings on the microtiter plate. Each plug of the plurality of plugs has a handling portion. The system includes a liquid handler including a gantry system including a plug-handling system configured to temporarily attach to respective handling portions of the plurality of plugs, and a pipetting system configured to dispense or aspirate liquids through the plurality of openings on the microtiter plate assembly without contacting the microtiter plate assembly.

Description

BIOREACTOR WELL ACCESSCROSS REFERENCE TO RELATED APPLICATION

[0001] This application is being filed on January 30, 2025, as a PCT International application and claims the benefit of and priority' to U.S. Provisional Patent Application No. 63 / 627,559, filed on January’ 31, 2024, the disclosure of which is hereby incorporated by reference in its entirety.BACKGROUND

[0002] Miniaturization and parallelization in the industrial production of microorganisms have gained economic importance in recent decades. In some examples, biological samples are screened for the selection of suitable biological strains, enzymes, or suitable culture media relevant to pharmacological and medicinal applications. In this context, there is a need for high sample throughputs which may be achieved via parallelization of experiments.

[0003] A microtiter plate is one example of a device that can be utilized to achieve a high number of parallel operations. Such devices typically have a flat plate with multiple wells that are used as small test tubes. Each of the individual wells may' be filled with a medium, inoculated to introduce cells into the medium, and incubated at a particular temperature using a shaking incubator. Process parameters, such as a pH value, concentrations of dissolved oxygen (DO), dissolved carbon dioxide, and biomass, and other parameter values may be continuously monitored for each individual well during the growth process.

[0004] The wells on the microtiter plate must be sealed for controlled gassing and prevention of contamination of the cell culture. Current methods of sealing microtiter plates typically involve a thick rubber pad that cannot be easily accessed by disposable tips, and is instead typically accessed by fixed tips which can increase the risk of contamination of the wells. Additional equipment and time for cleaning fixed tips are ty pically required to mitigate this increased risk of contamination, resulting in inefficiencies.SUMMARY

[0005] In general terms, the present disclosure relates to improved access to bioreactor wells. In one possible configuration, a microtiter plate assembly and a plug provide contactless dispense or aspiration of liquids into a bioreactor well. Variousaspects are described in this disclosure, which include, but are not limited to, the following aspects.

[0006] One aspect relates to a system for preparing microbial cultures, the system comprising: a microtiter plate assembly including: a plurality of openings; a plurality' of plugs sized and configured to be received within the plurality of openings on the microtiter plate, each plug of the plurality of plugs having a handling portion; and a liquid handler including: a gantry system including: a plug-handling system configured to temporarily attach to respective handling portions of the plurality of plugs; and a pipetting system configured to dispense or aspirate liquids through the plurality of openings on the microtiter plate assembly without contacting the microtiter plate assembly.

[0007] Another aspect relates to a microtiter plate assembly for a bioreactor, the microtiter plate assembly comprising: a first plate having a first plurality of openings; a second plate that aligns with the first plate, the second plate having channels for gas transport and a second plurality of openings that align with the first plurality of openings; and a plurality of plugs for sealing the first and second pluralities of openings in the first plate and second plate respectively, each plug of the plurality of plugs having: a handling portion configured for attachment to a pipetting system of a liquid handler.

[0008] Another aspect relates to a system for preparing microbial cultures, the system comprising: a microtiter plate assembly including: a first plate having a first plurality of openings; a second plate having channels for gas transport and a second plurality of openings that align with the first plurality of openings; and a plurality of plugs sealing the first and second pluralities of openings in the first and second plates, each plug of the plurality of plugs having: a body including a first tapered exterior surface forming a funnel lid geometry, the body defining a bore; and a core housed inside the bore of the body, the core forming a handling portion; and a liquid handler including: a gantry system including: a plug-handling system configured to temporarily attach to the handling portions of the plurality of plugs; and a pipetting system configured to dispense or aspirate liquids through the first and second pluralities of openings on the microtiter plate assembly without contacting the microtiter plate assembly.

[0009] A variety of additional aspects will be set forth in the description that follows. The aspects can relate to individual features and to combination of features. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the broad inventive concepts upon which the embodiments disclosed herein are based.DESCRIPTION OF THE FIGURES

[0010] The following drawing figures, which form a part of this application, are illustrative of the described technology and are not meant to limit the scope of the disclosure in any manner.

[0011] FIG. 1 is an isometric view of an example of a microbioreactor.

[0012] FIG. 2 is a front view of an example of a liquid handler integrated with the microbioreactor of FIG. 1.

[0013] FIG. 3 is an isometric view of an example of pipetting systems grouped together in a pipetting assembly of the liquid handler of FIG. 2.

[0014] FIG. 4 is an isometric view of an example of a pipetting system from the pipetting assembly of FIG. 3.

[0015] FIG. 5 is an isometric view of an attachment tube that is part of an attachment assembly shown in FIG. 4.

[0016] FIG. 6 is an isometric view of an example of a microtiter plate assembly of the microbioreactor of FIG. 1.

[0017] FIG. 7 is an exploded isometric view7of the microtiter plate assembly of FIG. 6.

[0018] FIG. 8 is another exploded isometric view of the microtiter plate assembly of FIG. 6.

[0019] FIG. 9 is an isometric cross-sectional view7of the microtiter plate assembly taken along the broken line shown in FIG. 6.

[0020] FIG. 10 is a front cross-sectional view of the microtiter plate assembly taken along the broken line shown in FIG. 6.

[0021] FIG. 11 is an isometric view of another example of the microtiter plate assembly of the microbioreactor of FIG. 1.

[0022] FIG. 12 is an isometric cross-sectional view7of the microtiter plate assembly taken along the broken line shown in FIG. 11.

[0023] FIG. 13 is a front cross-sectional view of the microtiter plate assembly taken along the broken line shown in FIG. 11.

[0024] FIG. 14 is an isometric view of a plug that can be used to seal an opening in the microtiter plate assemblies of FIGS. 6 and 11.

[0025] FIG. 15 is a side view of the plug of FIG. 14.

[0026] FIG. 16 schematically illustrates an example of a method of preparing microbial cultures using the liquid handler of FIG. 2 and the microtiter plate assemblies of FIGS . 6 and 11.DETAILED DESCRIPTION

[0027] Various embodiments will be described in detail with reference to the drawings, wherein like reference numerals represent like parts and assemblies throughout the several views. Reference to various embodiments does not limit the scope of the claims attached hereto. Additionally, any examples set forth in this specification are not intended to be limiting and merely set forth some of the manypossible embodiments for the appended claims.

[0028] FIG. 1 is an isometric view" of an example of a microbioreactor 100. As shown in the example of FIG. 1, the microbioreactor 100 includes a housing 102 that defines a cultivation chamber 104. The microbioreactor 100 monitors parameters such as biomass, pH, dissolved oxygen (DO), and fluorescence while running a cultivation inside the cultivation chamber 104. Additionally, the microbioreactor 100 includes a touchscreen display 106 that allows a user to control the shaking speed, temperature, gas concentration, gas flow rate, and humidity inside the cultivation chamber 104. Alternatively, or additionally, the microbioreactor 100 may be communicatively coupled to a separate computing device that may allow for such control.

[0029] In some aspects, the microbioreactor 100 can share similar components, features, and functionalities w ith the microbioreactor described in PCT International Patent Application No. PCT / US22 / 28207, titled Container Assembly for Microbioreactor, filed on May 6, 2022, the entirety of which is incorporated herein by reference.

[0030] FIG. 2 is a front view of an example of a liquid handler 200 integrated with the microbioreactor 100. Together, the microbioreactor 100 and the liquid handler 200 form a system 10 for preparing microbial cultures. As shown in the example of FIG. 2, the liquid handler 200 includes a pipetting assembly 300 for dispensing and aspiratingliquids into wells formed on a microtiter plate held inside the cultivation chamber 104 of the microbioreactor 100.

[0031] The pipetting assembly 300 includes a gantry system 302 that controls movement of a plug-handling system 304 and a pipetting system 305 about X, Y, and Z axes. In some aspects, the pipetting assembly 300 includes aspects of the pipetting assembly described in U.S. Provisional Patent Application No. 63 / 613,480, titled Replaceable Pipetting System, filed on December 21. 2023, the entirety of which is incorporated herein by reference.

[0032] FIG. 3 is an isometric view of an example of pipetting systems 305 grouped together in the pipetting assembly 300. In this example, eight pipetting systems are grouped together in the pipetting assembly 300. In alternative examples, more than eight pipetting systems or fewer than eight pipetting systems can be grouped together in the pipetting assembly 300.

[0033] As shown in FIG. 3, each pipetting system 305 includes a pipette tip 306. The pipetting systems 305 may move as a group or. in certain examples, individually. The pipetting systems 305 may be held together by a framework 312 about one or more axes, such that the pipetting systems 305 may move together about that those axes. For example, the framework 312 may hold the pipetting systems 305 together during movement about the X and Z axes. The pipetting systems 305 may then be free to move independently about the Y axis.

[0034] FIG. 4 is an isometric view of an example of a pipetting system 305. It is noted that the plug-handling system 304 is substantially similar to the pipetting system 305 such that the following description with respect to the pipetting system 305 in FIG. 4 can similarly apply to the plug-handling system 304. The plug-handling system 304 differs from the pipetting system 305 in that the pipette tip 306 is replaced with a modified pipette tip 308a, 308b.

[0035] As shown in FIG. 4, the pipetting system 305 includes a probe assembly 320 and an attachment assembly 340. The probe assembly 320 has s a gantry attachment structure 322 that attaches the probe assembly 320 to the pipetting assembly 300 such that the gantry system 302 is able to move the pipetting system 305 at least about the X and Z axes. In the example shown, the probe assembly 320 includes two motors, a first motor 324 and a second motor 326. The probe assembly 320 further includes a probeassembly housing 328 that houses a first rail and a second rail 330, 332 which are associated with first and second drive screws 334, 336.

[0036] FIG. 5 is an isometric view of an attachment tube 346 that is part of the attachment assembly 340 shown in FIG. 4. Referring now to FIGS. 4 and 5, the attachment assembly 340 includes an upper plunger housing 342 and a lower plunger housing 344. The attachment tube 346 extends at least partially through the upper and lower plunger housings 342. 344.

[0037] The attachment tube 346 extends about a pipetting axis between a first end 348 and a second end 350 and is radially outward of a plunger and an insulating tube about the pipetting axis. The attachment tube 346 is radially inward of the upper and lower plunger housings 342. 344 about the pipetting axis. The attachment tube 346 includes a first portion 352 and a second portion 354 at the pipette tip receiving region. The second portion 354 includes a first step 356 with an outer diameter that is smaller than an outer diameter of the first portion 352. The first step 356 is sized such that a first type of attachment (e.g., a first type of pipette tip) can slide and attach onto the outer diameter of the first step 356 such as by friction.

[0038] The second portion 354 also includes a second step 358 with a smaller outer diameter than that of the first step 356. The second step 358 is adjacent the first step 356, and extends to the second end 350 of the attachment tube 346. The second step 358 is sized such that a second type of attachment (e.g.. a second type of pipette tip) can slide and attach onto the outer diameter of the second step 358 such as by friction. An end of the second type of attachment is stopped by the transition between the first and second steps 356, 358.

[0039] The attachment tube 346 extends into an upper plunger tube of the upper plunger housing 342 about the pipetting axis. The attachment tube 346 also extends into a tubular structure of the lower plunger housing 344. In certain examples, the first and second steps 356, 358 extend out of the second end of the lower plunger housing 344 at least when the plunger is at the extended state, or between the extended state and the retracted state.

[0040] The attachment tube 346 is an electrically conductive tube. In certain examples, the attachment tube 346 is made of metal. In an assembled state, the attachment tube 346 is insulated by the insulating tube, such that the attachment tube 346 does not contact any conductive surfaces within the attachment assembly 340.

[0041] FIG. 6 is an isometric view of an example of a microtiter plate assembly 400 of the microbioreactor 100. The microtiter plate assembly 400 is housed inside the cultivation chamber 104 where orbital shaking is applied to the microtiter plate assembly 400 during cell cultivation. The microtiter plate assembly 400 includes a first plate 500, and a second plate 600 that aligns with the first plate 500. The second plate 600 has channels for gas transport to cell cultivation wells (not shown) which are positioned below the microtiter plate assembly 400.

[0042] FIG. 7 is an exploded isometric view of the microtiter plate assembly 400 from a top perspective. FIG. 8 is another exploded isometric view of the microtiter plate assembly 400 from a bottom perspective. In this illustrative example, the first plate 500 includes tabs 504 that are received inside apertures 604 on the second plate 600 for aligning the first plate 500 with the second plate 600. In alternative examples, the second plate 600 can include tabs that align with apertures on the first plate 500 for aligning the first plate 500 with the second plate 600.

[0043] The first plate 500 has a first plurality of openings 502, and the second plate 600 has a second plurality of openings 606 that align with the first plurality of openings 502 of the first plate 500. The first and second pluralities of openings 502, 606 provide access to cell cultivation wells (not shown) which are positioned below the microtiter plate assembly 400.

[0044] FIG. 9 is an isometric cross-sectional view of the microtiter plate assembly 400 taken along the broken line shown in FIG. 6. FIG. 10 is a front cross-sectional view of the microtiter plate assembly 400 taken along the broken line show n in FIG. 6. Referring now to FIGS. 6, 9, and 10, a plug 700 is used to seal the first and second openings 502, 606 in the first plate 500 and second plate 600. respectively. While the example provided in FIGS. 6, 9, and 10 shows a single plug, it is noted that the microtiter plate assembly 400 can include a plurality of plugs for sealing the openings on the microtiter plate assembly 400. As an illustrative example, the microtiter plate assembly 400 can include 24 plugs for sealing 24 openings on the microtiter plate assembly 400.

[0045] The plug 700 has a body 702 defining a bore 703. Additionally, the plug 700 includes a core 704 housed inside the bore 703. The core 704 forms a handling portion 706 that is configured for attachment to the plug-handling system 304 of the liquid handler 200, as will be described in more detail further below. In the example ofFIGS. 6, 9, and 10, the core 704 of the plug 700 extends at least partially outside of the bore 703 forming the handling portion 706. In alternative examples, the core 704 can be flush with a top surface of the plug 700.

[0046] In the example of FIGS. 6, 9, and 10, a modified pipette tip 308a can be attached to the second end 350 of the attachment tube 346 (see FIG. 5). The modified pipette tip 308a extends from a proximal end 309 to a distal end 311, where an aperture 313 on the proximal end 309 can fit around the first step 356 or the second step 358 of the attachment tube 346 such that the modified pipette tip 308a can be held on the attachment tube 346 by friction. In this example, a magnet 310 is inserted into an aperture 315 on the distal end 311 of the modified pipette tip 308a such that the magnet 310 is held on the modified pipette tip 308a by friction. Accordingly, the modified pipette tip 308a can also be referred to as a holder or gripper for the plug 700. In further examples, a geometry may be molded into the modified pipette tip 308a to positively locate and fit the magnet 310 into the aperture 315 on the distal end 311.

[0047] The body 702 of the plug 700 is made from a material that is stable across range of temperatures (e.g., up to 40°C), that is chemically resistant, and durable. For example, the body 702 can be made of a plastic material such as polypropylene or other similar types of plastic materials. The core 704 of the plug 700 is made from a ferromagnetic material that is attracted to the magnet 310. It is contemplated that the plug 700 is disposable after use.

[0048] The liquid handler 200 is programmed to control the gantry system 302 to lower a plug-handling system 304 such that the magnet 310 held on the modified pipette tip 308a is positioned proximate or otherwise abuts the handling portion 706 of the plug 700 such that when the gantry system 302 raises the plug-handling system 304, the plug 700 is removed from the openings 502, 606 in the first plate 500 and second plate 600 respectively.

[0049] The liquid handler 200 is further programmed to control the gantry7system 302 to lower a pipetting system 305 having a pipette tip 306 to dispense or aspirate a liquid into the cell cultivation wells (not shown) which are positioned below the microtiter plate assembly 400. Advantageously, the pipette tip 306 can enter the cultivation wells without contacting the microtiter plate assembly 400. This can reduce risk of contamination of the cell cultivation wells. Additionally, by not contacting the microtiter plate assembly 400. a broader range of pipette sizes can be used to dispenseand aspirate liquids into the cell cultivation wells. Also, disposable pipettes can be used to dispense and aspirate liquids into the cell cultivation wells to further reduce risk of contamination of the cell cultivation wells.

[0050] The liquid handler 200 is programmed to control the gantry system 302 to lower the plug-handling system 304 such that the plug 700 is inserted into the openings 502, 606 in the first plate 500 and second plate 600. respectively. Orbital shaking can be applied to the microtiter plate assembly 400 when housed inside the cultivation chamber 104 during cell cultivation. The orbital shaking can break the magnetic attachment between the magnet 310 of the plug-handling system 304 and the handling portion 706 of the plug 700 such that the plug 700 is shucked into the openings 502, 606 in the first plate 500 and second plate 600, respectively. Alternatively, lateral movement of the plug-handling system 304 relative to the microtiter plate assembly 400 may also break the magnetic attachment between the magnet 310 of the plughandling system 304 and the handling portion 706 of the plug 700. The plug 700 seals the cultivation wells positioned below the microtiter plate assembly 400 while allowing the pipetting assembly 300 to dispense or aspirate liquids without contacting the microtiter plate assembly 400.

[0051] FIG. 11 is an isometric view of another example of the microtiter plate assembly 400 and a modified pipette tip 308b that is positioned proximate the handling portion 706 of the plug 700. FIG. 12 is an isometric cross-sectional view of the microtiter plate assembly 400 and the modified pipette tip 308b taken along the broken line shown in FIG. 11. FIG. 13 is a front cross-sectional view of the microtiter plate assembly 400 and the modified pipette tip 308b taken along the broken line shown in FIG. 11. In the example shown in FIGS. 11-13, the microtiter plate assembly 400 and the plug 700 are the same as shown in the example of FIGS. 6, 9, and 10. The modified pipette tip 308b differs from the modified pipette tip 308a shown in FIGS. 6, 9, and 10 in that it does not include a magnet at the distal end 311.

[0052] Instead, as shown in FIG. 13, the aperture 315 on the distal end 311 of the modified pipette tip 308b has an interior diameter Di that corresponds with an outside diameter Do of the handling portion 706 of the plug 700. As an illustrative example, the outside diameter Do of the handling portion 706 can measure about 0. 1575 inches (0.40 cm), and the interior diameter Di of the modified pipette tip 308b can measure about 0. 1552 inches (0.39 cm).

[0053] When the liquid handler 200 controls the gantry system 302 to lower the plug-handling system 304. the handling portion 706 is received inside the aperture 315. A frictional force between the interior diameter Di of the modified pipette tip 308b and the outside diameter Do of the handling portion 706 causes the plug-handling system 304 to temporarily attach to the handling portion 706 of the plug 700 such that when the gantry system 302 raises the plug-handling system 304, the plug 700 is removed from the openings 502, 606 in the first plate 500 and second plate 600 respectively. Accordingly, the modified pipette tip 308b can also be referred to as a holder or gripper for the plug 700.

[0054] While the plug 700 is removed from the openings 502, 606 in the first plate 500 and second plate 600, the liquid handler 200 controls the gantry system 302 to lower the pipetting system 305 having the pipette tip 306 to dispense or aspirate a liquid into the cell cultivation wells. Like in the example described above with respect to FIGS. 6, 9, and 19, the pipette tip 306 can enter the cultivation wells without contacting the microtiter plate assembly 400. This can reduce risk of contamination of the cell cultivation wells. Additionally, by not contacting the microtiter plate assembly 400, a broader range of pipette sizes can be used to dispense and aspirate liquids into the cell cultivation wells. Also, disposable pipettes can be used to dispense and aspirate liquids into the cell cultivation wells to further reduce risk of contamination of the cell cultivation wells.

[0055] The liquid handler 200 is programmed to control the gantry system 302 to lower the plug-handling system 304 such that the plug 700 is inserted into the openings 502, 606 in the first plate 500 and second plate 600, respectively. The orbital shaking applied to the microtiter plate assembly 400 during cell cultivation can break the frictional force between the interior diameter Di of the modified pipette tip 308b and the outside diameter Do of the handling portion 706 such that the plug 700 is shucked into the openings 502, 606 in the first plate 500 and second plate 600, respectively. Alternatively, lateral movement of the plug-handling system 304 relative to the microtiter plate assembly 400 may also break the frictional force between the interior diameter Di of the modified pipette tip 308b and the outside diameter Do of the handling portion 706 of the plug 700. In this manner, the plug 700 seals the cell cultivation wells positioned below the microtiter plate assembly 400 while allowing thepipeting assembly 300 to dispense or aspirate liquids without contacting the microtiter plate assembly 400.

[0056] In a further alternative example, the plug 700 can omit the core 704 such that the bore 703 is empty. In such examples, the second end 350 of the atachment tube 346 has a geometry that fits inside the bore 703. A frictional force between the second end 350 of the atachment tube 346 and the interior walls of the bore 703 provides a temporarily atachment allowing the plug-handling system 304 to position the plug 700 inside the openings 502, 606 in the first plate 500 and second plate 600, and to remove the plug 700 from the openings 502, 606 in the first plate 500 and second plate 600, as discussed in the examples described above.

[0057] FIG. 14 is an isometric view of the plug 700. FIG. 15 is a side view of the plug 700. Referring now to FIGS. 14 and 15, the plug 700 includes a first cylindrical exterior surface 708 defining a first outside diameter Doi. As an illustrative example, first outside diameter Doi of the first cylindrical exterior surface 708 can measure about 0.56 inches (1.42 cm).

[0058] The plug 700 further includes a first tapered exterior surface 710 that extends from the first cylindrical exterior surface 708. The first tapered exterior surface 710 forms a funnel lid geometry'. The first tapered exterior surface 710 corresponds to a tapered perimeter 506 that surrounds the openings 502 in the first plate 500 (see FIGS. 7, 9, and 12). The first tapered exterior surface 710 on the plug 700 and the tapered perimeters 506 around each opening 502 facilitate the shucking of the plug 700 into the openings 502, 606 in the first plate 500 and second plate 600 respectively by providing a tolerance that minimizes motions (e.g., in X-axis direction and in Y-axis direction) due to the orbital shaking of the microtiter plate assembly 400.

[0059] The plug 700 further includes a sealing surface 714 for receiving a seal. An example of the seal can include a gasket such as an O-ring or similar ty pe of seal. The seal can be made of rubber or similar material. The seal when received in the sealing surface 714 seals the gases transported by the second plate 600 to the cell cultivation wells that are positioned below.

[0060] The plug 700 further includes a second cylindrical exterior surface 712 defining a second outside diameter D02, and a third cylindrical exterior surface 716 defining a third outside diameter D03. The sealing surface 714 is positioned between the second and third cylindrical exterior surfaces 712. 716. As shown in FIG. 15. thethird outside diameter D03 is less than the second outside diameter D02, and the second outside diameter D02 is less than the first outside diameter Doi. Further, the plug 700 includes a second tapered exterior surface 718. In some examples, the second tapered exterior surface 718 at least partially enters a cell cultivation well positioned below the microtiter plate assembly 400 for sealing the cell cultivation well.

[0061] As an illustrative example, the second outside diameter D02 of the plug 700 can measure about 0.4165 inches (1.06 cm). As another illustrative example, the third outside diameter D03 of the plug 700 can measure about 0.36 inches (0.91 cm).

[0062] FIG. 16 schematically illustrates an example of a method 1600 of preparing microbial cultures using the liquid handler 200 and the microtiter plate assembly 400. The method 1600 includes an operation 1602 of removing a plug 700 from the openings 502, 606 in the first plate 500 and second plate 600. Operation 1602 can include controlling the gantry system 302 to lower a plug-handling system 304 such that the modified pipette tip 308a, 308b temporarily attaches to the handling portion 706 of the plug 700. Operation 1602 can further include controlling the gantry system 302 to raise the plug-handling system 304 such that the plug 700 is removed from the openings 502, 606 in the first plate 500 and second plate 600, respectively.

[0063] The method 1600 includes an operation 1604 of inserting a pipette tip 306 through the openings 502, 606 in the first plate 500 and second plate 600. In some examples, operation 1604 further includes inserting the pipette tip 306 into a cell cultivation well below the microtiter plate assembly 400. Operation 1604 can include controlling the gantry system 302 to lower a pipetting system 305 to have the pipette tip 306 inserted into openings 502, 606 in the first plate 500 and second plate 600, and optionally into a cell cultivation well below the microtiter plate assembly 400. Advantageously, operation 1604 can be performed without touching the microtiter plate assembly 400 such that the operation 1604 is contactless. By eliminating contact with the microtiter plate assembly 400, contamination of the cell cultivation well is reduced, and a larger variety' of pipette tips 306 can be used such as disposable pipette tips ranging in different sizes.

[0064] The method 1600 includes an operation 1606 of performing an action using the pipette tip 306. For example, operation 1606 can include dispensing a liquid into the cell cultivation well, or can include aspirating a liquid from the cell cultivation well.

[0065] The method 1600 includes an operation 1608 of removing the pipette tip 306 from the openings 502, 606 in the first plate 500 and second plate 600. respectively. In examples where operation 1604 includes inserting the pipette tip 306 into the cell cultivation well, operation 1608 can include removing the pipette tip 306 from the cell cultivation well. Operation 1608 can include controlling the gantry system 302 to raise the pipetting system 305 to have the pipette tip 306 removed from the openings 502, 606 in the first plate 500 and second plate 600 and from the cell cultivation well below the microtiter plate assembly 400. Advantageously, operation 1608 can be performed without touching the microtiter plate assembly 400 such that the operation 1608 is contactless. As discussed above, by eliminating contact with the microtiter plate assembly 400. contamination of the cell cultivation well is reduced, and a larger variety of pipette tips 306 can be used such as disposable pipette tips ranging in different sizes.

[0066] The method 1600 includes an operation 1610 of inserting the plug 700 back into the openings 502, 606 in the first plate 500 and second plate 600, respectively. Operation 1610 can include controlling the gantry system 302 to lower the plughandling system 304 such that the plug 700 is inserted into the openings 502, 606 in the first plate 500 and second plate 600, respectively. The orbital shaking breaks the temporary attachment between the modified pipette tip 308a, 308b and the handling portion 706 of the plug 700 such that the plug 700 is shucked into the openings 502, 606 in the first plate 500 and second plate 600, respectively. Alternatively, lateral movement of the plug-handling system 304 relative to the microtiter plate assembly 400 may also break the temporary attachment between the modified pipette tip 308a. 308b and the handling portion 706 of the plug 700 such that the plug 700. After completion of operation 1610, the plug 700 seals the cultivation wells positioned below the microtiter plate assembly 400.

[0067] The various embodiments described above are provided by way of illustration only and should not be construed to be limiting in any way. Various modifications can be made to the embodiments described above without departing from the true spirit and scope of the disclosure.

Claims

What is claimed is:

1. A system for preparing microbial cultures, the system comprising: a microtiter plate assembly including: a plurality of openings; a plurality of plugs sized and configured to be received within the plurality of openings on the microtiter plate, each plug of the plurality of plugs having a handling portion; and a liquid handler including: a gantry system including: a plug-handling system configured to temporarily attach to respective handling portions of the plurality of plugs; and a pipetting system configured to dispense or aspirate liquids through the plurality of openings on the microtiter plate assembly without contacting the microtiter plate assembly.

2. The system of claim 1 , wherein the plug-handling system includes magnets that magnetically attach to the handling portions of the plurality' of plugs.

3. The system of claim 2, wherein magnetic attachment between the plug-handling system and the handling portions of the plurality of plugs is broken by relative motion between the plug-handling system and the microtiter plate assembly.

4. The system of claim 1, wherein the plug-handling system includes holders each having an inside diameter that corresponds with an outside diameter of the handling portions of the plurality of plugs such that the holders attach to the handling portions by friction.

5. The system of claim 4, wherein attachment between the holders of the plug-handling system and the handling portions of the plurality of plugs is broken by relative motion between the plug-handling system and the microtiter plate assembly.

6. The system of any of claims 1-5, wherein each plug of the plurality of plugs has a body defining a bore, and a core housed inside the bore.

7. The system of claim 6, wherein the body is made from a plastic material, and the core is made from a ferromagnetic material.

8. The system of any of claims 1-7, wherein the core of each plug of the plurality of plugs extends at least partially outside of the bore forming the handling portion9. The system of any of claims 1-8, wherein each plug of the plurality' of plugs includes a sealing surface for receiving a seal.

10. The system of any of claims 1-9, wherein each plug of the plurality of plugs further includes a first tapered exterior surface forming a funnel lid geometry'.

11. The system of any of claims 1-9, wherein the liquid handler includes a processing circuitry having a memory for storing instructions which, when executed by the processing circuitry', cause the processing circuitry' to: control the gantry system to lower the plug-handling system to temporarily attach to the handling portions of the plurality of plugs; control the gantry system to raise the plug-handling system such that the plurality of plugs are removed from the plurality of openings; control the gantry' system to lower the pipetting system to have pipette tips inserted into the plurality of openings without touching the microtiter plate assembly; perform an action using the pipette tips; control the gantry system to raise the pipetting system to have the pipette tips removed from the plurality of openings without touching the microtiter plate assembly; and control the gantry system to lower the plug-handling system such that the plurality of plugs are inserted into the plurality of openings.

12. A microtiter plate assembly for a bioreactor, the microtiter plate assembly comprising:a first plate having a first plurality of openings; a second plate that aligns with the first plate, the second plate having channels for gas transport and a second plurality of openings that align with the first plurality of openings; and a plurality of plugs for sealing the first and second pluralities of openings in the first plate and second plate respectively, each plug of the plurality of plugs having: a handling portion configured for attachment to a pipetting system of a liquid handler.

13. The microtiter plate assembly of claim 12, wherein each plug further includes: a body defining a bore; and a core housed inside the bore, wherein the body of each plug is made from a plastic material, and wherein the core of each plug is made from a ferromagnetic material.

14. The microtiter plate assembly of claim 13, wherein the core of each plug extends at least partially outside of the bore forming the handling portion.

15. The microliter plate assembly of any of claims 12-14, wherein each plug includes a sealing surface for receiving a seal.

16. The microtiter plate assembly of any of claims 12-15, wherein each plug further includes a first tapered exterior surface forming a funnel lid geometry.

17. A system for preparing microbial cultures, the system comprising: a microtiter plate assembly including: a first plate having a first plurality of openings; a second plate having channels for gas transport and a second plurality of openings that align with the first plurality of openings; and a plurality of plugs sealing the first and second pluralities of openings in the first and second plates, each plug of the plurality of plugs having: a body including a first tapered exterior surface forming a funnel lid geometry, the body defining a bore; anda core housed inside the bore of the body, the core forming a handling portion; and a liquid handler including: a gantry system including: a plug-handling system configured to temporarily attach to the handling portions of the plurality of plugs; and a pipetting system configured to dispense or aspirate liquids through the first and second pluralities of openings on the microtiter plate assembly without contacting the microtiter plate assembly.

18. The system of claim 17, wherein the body is made from a plastic material, and wherein the core is made from a ferromagnetic material.

19. The system of claim 17 or 18, wherein the core extends at least partially outside of the bore forming the handling portion.

20. The system of any of claims 17-19, further comprising: a sealing surface for receiving a seal.

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