Automated orbital culture shaker compartment

Abstract

The present invention relates to a set for maintaining and transporting at least one cell culture flask placed in at least one cell culture flask holder (2, 12), and to methods and uses of said set.The present invention relates to an orbital culture shaker comprising a set (7) of the present invention, and to a cell culture flask holder (2, 12) compatible with said set.

Description

[Technical field]

[0001] The present invention relates to a cell culture platform, in particular an automated cell culture platform comprising a shaking incubator and a means for handling laboratory equipment in the automated cell culture platform. In particular, the present invention relates to an automated culture shaker compartment for use in the automated culture platform. [Background technology]

[0002] Cell culture refers to laboratory methods that allow for the growth of eukaryotic or prokaryotic cells in physiological conditions. It is a method used, for example, to study tissue growth and maturation, viral biology and vaccine development, the role of genes in disease and health, and the use of large-scale hybrid cell lines to generate biopharmaceuticals. The experimental uses of cultured cells are as diverse as the cell types that can be grown in vitro. One particular example of a cell culture method is cell passaging (or subculture), which involves the removal of medium and the transfer of cells from the previous culture into fresh growth medium. This procedure allows for further propagation of the cell line or cell strain.

[0003] Cell culture methods require the use of dedicated laboratory equipment and precise multi-step cell manipulations. Manual procedures are difficult to adapt to GMP (Good Manufacturing Practice) or GLP (Good Laboratory Practice) facilities and therefore they are not usually used in biotech or pharmaceutical companies. Therefore, some processes / steps of cell culture are automated. One example of an automated cell passaging platform is the microtiter plate (MTP) culture format, as reported by the University of Luxembourg (Automated Cell Culture Maintenance and High Throughput / High Content Screening Platform; https: / / wwwen.uni.lu / lcsb / research / translational_neuroscience / automation_platform.). This is an example of a high-throughput / high-content platform for maintaining induced pluripotent stem cells in culture and differentiating these cells into neurons while performing chemical or viral vector-based genetic screens.

[0004] One example of an apparatus used for cell culture is the shaking (or shaker) incubator, which is a laboratory device that allows the incubation and shaking of cell cultures. A typical culture shaker includes at least a chamber with a door, a motor and shaking control means, and a shaking tray with temperature control means. The loading and unloading of the shaking tray is usually done manually and involves the opening of the door. To further reduce the manual operation of the shaking labware, the process can be automated such that the opening and closing of the incubator door is automated and a robotic arm located outside the incubator can perform the loading and unloading of the shaking tray. The automated process is usually compatible with one type of labware, typically a microtiter plate (MTP). In this example, the loading and unloading of the shaking tray needs to be done when the door is open, and the robotic arm mimics the manual actions of, for example, manipulating the MTP. Cell culture in the incubator requires the maintenance of temperature, humidity and composition of a defined gas atmosphere in the incubator. If the incubator is open during culture, the temperature, humidity and composition of the gas atmosphere within the incubator, and therefore within the culture, can change and adversely affect the cells.

[0005] Furthermore, shaking incubators usually include a universal tray that can be fitted with a tray clamp for labware such as clamps for Erlenmeyer flasks, test tube holders, microtiter plates, or other custom holders. In another example, such a tray is equipped with pins, whereby flasks are held in place by these pins. In another example, such a tray is equipped with adhesive strips, whereby labware is placed directly on the adhesive strips without the need for any other support. Most of this solution is not compatible with the mounting and removal of the shaking tray being performed by an automated robotic arm.

[0006] It is an object of the present invention to reduce the manual manipulation of shaking cultures and to provide improved devices and methods for the handling of cell cultures contained in laboratory equipment suitable for automated culturing and cell culture.

[0007] Document CN112940917 discloses a robot for the manipulation of culture bottles. These culture bottles are in the shape of Petri dishes and are moved individually by the arm of the robot into a frame, said frame being attached to a device capable of rotating and vibrating. Document JP 2009-284810 describes an incubator equipped with a shutter and a heated conveyor for transporting containers containing culture samples maintained in a static environment growth. A robot can move the containers into or out of the heated chamber while preventing changes in the culture conditions. Summary of the Invention

[0008] The aforementioned documents do not provide any address for the manipulation of suspension cells cultured in individual Erlenmeyer flasks or conical tubes mounted in flask or tube holders suitable for orbital movement within an orbital shaking incubator.

[0009] The present invention is based on the selection of preferred laboratory equipment for use in a cell passaging method for maintaining cell cultures, preferably for cell cultures in suspension. In particular, Erlenmeyer flasks or bioreactor tubes are suitable laboratory equipment for maintaining suspension cell cultures. A particularity of suspension cultures is the need to be stirred to avoid cell sedimentation to the bottom of the flask. While adherent cell cultures can be maintained in flat-bottomed stationary flasks with a large surface area (to promote cell adhesion), suspension cultures require stirring, otherwise the cells will settle to the bottom of the flask, greatly affecting their access to nutrients and oxygen, and ultimately resulting in cell death. For this reason, dedicated flasks (including, for example, shaker flasks and spinner flasks) are used to stir the medium and keep the cells in suspension. These flasks have in common a circular base area that allows a vortex-like motion to be generated by the orbital motion of the shaker.

[0010] The invention is based on providing a holder (or basket) (2, 12) for maintaining specialized labware containing cell cultures (e.g., flasks (1) as shown in Figure 1 or bioreactor tubes (11) as shown in Figure 9). The flasks have a circular bottom area and are designed for suspension cell culture. Cell suspension cultures simply grow single cells at high speed in a liquid medium. The liquid medium is continuously stirred on an orbital shaker. This culture method is used by scientists for cell growth research and development. The invention is based on providing an automated set (7) for maintaining and transporting cell culture labware (e.g., flasks, bioreactor tubes) that is suitable for an orbital culture shaker compartment and includes a culture shaker tray (3), a three-axis motorized stage (4), a sliding door (5), and a transport shuttle (6) (Figure 7). The invention relates to an orbital shaking incubator with an internal space, in which multiple baskets for suspension cell culture labware (e.g., flasks, bioreactor tubes) can be placed on the shaking tray. The orbital shaking incubator has a chamber designed to accommodate a set of holding and transporting cell culture labware (e.g., flasks, bioreactor tubes) holders. The present invention is particularly adapted for use in automated cell passaging platforms.

[0011] The present invention allows for the manipulation of suspension cell cultures containing labware individually and independently of each other in an orbital shaking incubator via a cell culture labware holder that is removably connected to a shaking tray. The present invention is designed for suspension cultures. Cell suspension cultures or suspension cultures are a type of cell culture that allows cells to grow in a growth medium that is agitated, thus forming a suspension.

[0012] In the case of parallel cultivation of multiple cultures in one orbital shaking incubator, one of the flask holders is taken out of the incubator via the set of the present invention, and this movement has only limited impact on the environment of the cultures in the other labware. Although the temperature changes slightly upon opening of the automated sliding door of the set, this small temperature change has minimal or no impact on the cultures in the other labware. The set of the present invention facilitates the installation and removal of the shaking tray, thus enhancing the ergonomic handling of the cultures.

[0013] In one embodiment, a set for maintaining and transporting at least one cell culture flask is provided, the set designed to be placed in an orbital culture shaker chamber, the set including: a flask holder, designed to receive at least one cell culture flask having a circular bottom surface and suitable for suspension cell culture; a culture shaker tray designed to be mounted inside an orbital culture shaker chamber, comprising at least one guiding member arranged to maintain each cell culture flask holder; an orbital culture shaker tray for receiving the flask holder and for transmitting orbital motion to the flask holder; - a three-axis motorized stage designed to be mounted inside the top wall of an orbital culture shaker chamber, gripping the X-axis, Y-axis and Z-axis, and three linear motors for driving the three axes and a cell culture flask holder; a sliding door designed to be mounted within a side wall of an orbital culture shaker chamber, the sliding door including a door panel and at least one motor for driving door movement; - a transport shuttle designed to be mounted on the outside of a side wall of an orbital culture shaker chamber, the transport shuttle including a slide frame, rails, at least two belts disposed on the slide frame, a cell culture flask holder plate and a motor for driving the sliding movement of the plate on the rails.

[0014] In another embodiment, a set for maintaining and transporting at least one suspension cell culture flask according to the present invention is provided, wherein the cell culture flask is a 125 ml Erlenmeyer flask or a bioreactor tube with a circular base area.

[0015] In another embodiment, a set according to the invention is provided, wherein the means for gripping the cell culture flask holder comprises one parallel gripper with adjustable gripping force mounted on the Z-axis of a three-axis motorized stage and with jaws and fingers for removable connection to the flask holder.

[0016] In another embodiment there is provided a set according to the invention, wherein the motor for driving the door movement of the sliding door comprises a stepper motor for driving the sliding movement and a stepper motor for driving the pressing movement of the door panel.

[0017] In another embodiment, there is provided a set according to the present invention, wherein the sliding door further comprises a precision sensor for detecting a sliding movement of the door panel and a precision sensor for detecting a pressing movement of the door panel.

[0018] In another embodiment, there is provided a set according to the present invention, wherein the sliding door further comprises a temperature probe, a heating pad, a heating ribbon and a controller, wherein the temperature probe controls the heating pad and the heating ribbon via the controller.

[0019] In another embodiment, there is provided a set according to the present invention, wherein the sliding door further comprises a silicone O-ring mounted between the door frame and the door panel.

[0020] In another embodiment, a set according to the present invention is provided, wherein the transport shuttle further comprises an amplified precision sensor for detecting a home position of the transport shuttle, a sensor for detecting an attachment position of the cell culture flask holder, and a sensor for detecting a removal position of the cell culture flask holder.

[0021] In another embodiment, a set according to the present invention is provided, further comprising a cell culture flask holder comprising a bottom surface, two handles on the bottom surface arranged perpendicular to the bottom surface and opposite each other, a locking means for holding the cell culture flask, and a heat transfer pad arranged on the bottom surface of the holder.

[0022] In another embodiment there is provided a set according to the invention, wherein the cell culture flask holder further comprises at least two magnets disposed on the exterior of a bottom surface of the cell culture flask holder providing a means for attachment to a culture shaker comprising a permanent electromagnet.

[0023] In another embodiment there is provided a set according to the invention, wherein the locking means of the cell culture flask holder comprises a clamping means and a safety latch located on a bottom surface of the cell culture flask holder.

[0024] In another embodiment a set according to the invention is provided, wherein the locking means of the cell culture flask holder comprises four guillotines, each holding a toothed jaw, and arranged on a bottom surface of the cell culture flask holder.

[0025] In another embodiment, there is provided a use of the set according to the invention for maintaining and transporting at least one cell culture flask disposed in at least one cell culture flask holder compatible with the set.

[0026] In another embodiment, a culture shaker is provided comprising a set according to the invention.

[0027] In another embodiment, an automated cell passaging platform is provided, comprising a liquid handling system, at least one robotized arm, at least one automated hotel, an automated refrigerated incubator, an automated microplate centrifuge with an automated loader, a cell counter and at least one orbital shaking incubator, and optionally the automated orbital shaking incubator, characterized in that at least one orbital shaking incubator is designed to be arranged in a set for maintaining and transporting at least one cell culture flask according to the present invention.

[0028] The aforementioned documents (CN112940917 and JP2009-284810) do not provide any address for the manipulation of suspension cells cultured in individual Erlenmeyer flasks or conical tubes mounted in flask or tube holders suitable for orbital movement in an orbital shaking incubator. [Brief description of the drawings]

[0029] [Figure 1] Figure 2 shows an example of a flask basket for a 125 ml Erlenmeyer flask; A, D, E show diagrams and panels B, C show photographs. The flask basket (2) for a 125 ml Erlenmeyer flask (1) includes a bottom (2.1), two handles (one handle-2.2), a heat transfer pad (2.3), a toothed spring with a horseshoe shape (2.4), a safety latch (2.5), a magnetic latch (2.6), a latch embossment (2.7), a counter magnetic pad (2.8), and an RFID tag or QR code (2.9). [Diagram 2]An example of a culture shaker tray (3) is shown. Panel A shows the lower plate (3.1) with 80 permanent electromagnets (3.4). Panel B shows the assembled lower plate (3.1) with the upper plate (3.2) (tray locker 3.3 not shown). The upper plate (3.2) has 80 locating pins (3.5) and 40 RFID antenna holes (3.6). Panel C shows the culture shaker tray (3) with 40 flask holders (2) for 125 ml Erlenmeyer flasks (1) arranged in an 8x5 array. [Diagram 3] An example of a three-axis motorized stage (4) is shown, which includes an X-axis (4.1), a Y-axis (4.2), a Z-axis (4.3), a linear motor (4.4) (not visible) for driving the X-axis (4.1), a linear motor (4.5) (not visible) for driving the Y-axis (4.2), a linear motor (4.6) (not visible) for driving the Z-axis (4.3), and a parallel gripper (4.7) with jaws (4.8) and fingers (4.9). [Figure 4] An example of a sliding door (5) is shown. Panel A: An example of a sliding door (5) including the door frame (5.1) and door panel (5.2), door tunnel (5.3), sliding motion driving stepper motor (5.4) and pressure motion driving stepper motor (5.5), precision sensor for sliding motion (5.6) and pressure motion (5.7), temperature probe (5.8) (not visible), heating pad (5.9) (not visible), O-ring (5.10) (not visible; in Figure 4D or 6C, marked) and heating ribbon (5.11) (not visible). Panels B-D show three parts of the door panel (5.2), which are the front panel (5.2.1) (Figure 4B shows the exterior view of the opened sliding door), the main body (5.2.2) (Figure 4C shows the center part of the opened sliding door) and the pressure panel (5.2.3) (Figure 4D). [Diagram 5]An example of a transport shuttle (6) is shown, which includes a sliding frame (6.1) with two upper and two lower rails (6.2), a sliding motion driving stepper motor (6.3), a pinion (6.4) (not visible), a flask holder plate (6.5) with two positioning pins (6.6) (not visible), an amplified precision sensor for detecting the transport shuttle home position (6.7), a sensor for detection of the flask holder attachment position (6.8) and a sensor for detection of the flask holder removal position (6.9), a presence laser detector (6.10), a belt (6.11) (not visible; marked in Figure 6A), a two-stage extension plate (6.12) and a ball screw (6.13) (not visible). [Figure 6] An example of a transport shuttle (6) designed to be attached to the outside of the sidewall of an orbital culture shaker chamber below the sliding door (5) is shown. Panel A shows the belt (6.11) of the transport shuttle (6) and panel C shows the O-ring (5.10) of the sliding door (5). [Figure 7] Shown is an example of a set (7) for maintaining and transporting cell culture flasks designed to be mounted in an orbital culture shaker chamber (chamber is marked as a shaded outline), which includes a culture shaker tray (3), a three-axis motorized stage (4), a sliding door (5), and a transport shuttle (6). [Figure 8] An example of the arrangement of a culture shaker (8) including a set (7) of the invention in an automated cell passaging platform is shown, where the automated orbital shaking incubator is integrated with a liquid handling system (9) and connected via a transport shuttle (6) through a hole / window drilled on the wall of the back cabinet (not marked) (Panel A). Panel B shows the sliding door (5) and transport shuttle (6) found on the outside of the side wall of the culture shaker for transporting flask holders to the liquid handling system (9). [Figure 9]FIG. 1 shows an example of a flask basket (12) for 50 ml bioreactor tubes (11) with locking means for four bioreactor tubes (10). Panels A and B show the diagram. The flask basket (12) for 50 ml bioreactor tubes includes four guillotines (10.1), eight toothed jaws (10.2), four rails and pivots (10.3) (pivots not visible), plunger (10.4), plunger holder (10.5), eccentric jig (10.6), control latch (10.7) and an RFID tag or QR code (2.9) (not marked).

[0030] Explanation of symbols 1. Flask (e.g., 125 ml Erlenmeyer flask); 1.1. Cap (e.g., 125 ml Erlenmeyer flask cap); 2. Flask basket; 2.1. Flask basket bottom; 2.2. Flask basket handle; 2.3. Heat transfer pad for flask basket; 2.4-2.7. Locking means for Erlenmeyer flask; 2.4. Tooth spring with horseshoe shape for flask basket; 2.5. Safety latch; 2.6. Magnetic latch; 2.7. Latch embossment; 2.8. Counter magnetic pad; 2.9. RFID tag or QR code; 3. incubator tray;3.1. bottom plate;3.2. top plate;3.3. tray locker;3.4. permanent electromagnet;3.5. locating pin;3.6. RFID antenna hole;4. three-axis motorized stage;4.1. X-axis;4.2. Y-axis;4.3. Z-axis;4.4. linear motor for X-axis (4.1);4.5. linear motor for Y-axis (4.2);4.6. linear motor for Z-axis (4.3);4.7. parallel gripper;4.8. parallel gripper jaws;4.9. parallel gripper fingers;5. sliding door;5.1. door frame;5.2. door panel;5.2.1.Front panel of door panel;5.2.2.Door panel body;5.2.3.Pressure panel of door panel;5.3.Door tunnel;5.4.Stepping motor driving sliding motion;5.5.Stepping motor driving pressure motion;5.6.Precision sensor for sliding motion;5.7.Precision sensor for pressure motion;5.8.Temperature probe;5.9.Warming pad;5.10.O-ring;5.11.Heating ribbon;6.Transport shuttle;6.1.Sliding frame;6.2.Upper and lower rails;6.3.Stepping motor driving sliding motion;6.4.Pinion 6.5. Flask holder plate; 6.6. Two positioning pins on the flask holder plate; 6.7. Amplified precision sensor for detecting the home position of the transport shuttle; 6.8. Sensor for detecting the mounting position of the flask holder; 6.9. Sensor for detecting the removal position of the flask holder; 6.10. Presence laser detector; 6.11. Belt; 6.12. Two-stage extension plate; 6.13. Ball screw; 7. Set for maintaining and transporting at least one cell culture flask; 8. Culture shaker comprising the set of the present invention; 9.Liquid handling system;10.1-10.7. Locking means for four bioreactor tubes;10.1. Guillotine;10.2. Toothed jaws;10.3. Rail and pivot;10.4. Plunger;10.5. Plunger holder;10.6. Eccentric jig;10.7. Control latch;10.8. Side heat conductor;10.9. Bottom heat conductor;10.10. Counter magnetic pads;11. Bioreactor tubes;11.1. Bioreactor tube caps (e.g. filter caps);12. Flask basket for bioreactor tubes. . DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0031] The term "orbital shaking incubator" or "orbital shaker incubator" or "orbital incubator shaker" (terms used interchangeably) refers to a laboratory device that combines incubation and orbital shaking functions. A typical orbital culture shaker includes at least a chamber (preferably made of stainless steel or painted stainless steel), a door (herein, the main door) including a door glass and a frame, a tray, a motor that moves the tray in a shaking motion, shaking control means (e.g., speed, frequency), heating / cooling means and means for temperature (heating / cooling) control. In addition, the culture shaker includes a means for regulating humidity and a means for controlling humidity, CO 2 Means for regulating the levels and CO 2The shaker may include means for temperature control as well as other environmental parameter (e.g., light) control. Culture shakers can be used for a variety of applications in the fields of biology, chemistry, biochemistry, pharmacology, etc., including but not limited to any type of cell culture (bacterial culture, tissue culture, and yeast), protein expression, solubility testing, or general mixing. Depending on the laboratory setting and the designated application, the shaker incubators range from compact tabletop to large capacity floor stackable types, etc., with various means for temperature control and means for shaking speed control. This application is developed around an orbital culture shaker that has a tray for receiving flask holders and transmitting orbital motion to the flask holders, where the orbital shaking motion is at a low speed (25-500 rpm).

[0032] The term "flask" or "shake flask" refers to a specific type of flask for suspension culture suitable for orbital shakers. These flasks have in common a circular base area where the orbital (or circular) motion increases the surface area of ​​the top surface of the liquid, maximizing gas exchange between the liquid and the air. A shake flask may be an Erlenmeyer flask, also known as a conical flask or titration flask, which features a flat bottom, a conical body, and a cylindrical neck. Another example of a shake flask may be a Fernbach flask, which has a baffle plate on the bottom to maximize oxygen transfer to the medium when shaken. Another example of a shake flask may be a bioreactor tube, which may have a conical bottom and allow for a smaller working volume of 1-15 mL.

[0033] The term "suspension cell culture" refers to a process whereby cells are grown under controlled conditions outside their natural environment. Cell suspension culture or suspension culture is a type of cell culture that allows cells to grow in an agitated growth medium, thus forming a suspension. Cells cultured in the laboratory can be classified into three different types: primary cells, transformed cells, and self-renewing cells. Common operations performed on cultured cells are changing the medium, passaging the cells, and transfecting the cells with genes. "Passaging" (also known as subculture or cell division) involves transferring a small number of cells into a new container. Cells can be cultured for longer times if they are divided regularly, as it avoids the senescence associated with long-term high cell densities.

[0034] Cell passaging may be performed by use of an automated cell passaging platform. An exemplary automated cell passaging platform may include: - Liquid handling systems (e.g. Hamilton VANTAGE 2.0 Liquid Handling System). - at least one robotic arm (e.g. Precise, PF400 extended), -At least one automated hotel (e.g. Liconic LPX740), - an automated refrigerated incubator (e.g. Liconic STX44-HCBT), Optionally, an automated ultra-low temperature freezer (e.g. Liconic STX88-DFMISA), - an automated microplate centrifuge with an automated loader (e.g. Agilent), - a cell counter (e.g., Raman Rxn2-785 Analyzer, Kaiser / Endress&Hauser), and - At least one (maximum four) orbital shaking incubators (e.g. Kuhner, ISF1-ZC Peltier).

[0035] Flask holder The present invention is based on the selection of preferred labware for maintaining cell cultures, preferably for use in cell passaging methods for seed train culture. In particular, Erlenmeyer flasks are preferred labware for maintaining cell cultures. Preferably, the 125 mL Erlenmeyer flask format provides sufficient amounts of cell culture for the required cell culture application (20 mL). However, Erlenmeyer flasks are considered tall labware, such as taller than microplates. The size of Erlenmeyer flasks is not compatible with known automated labware. However, Erlenmeyer flasks limit the risk of cross contamination compared to deep-bottom plate formats. Preferably, Erlenmeyer flasks are sterile, baffled and threaded flasks with 0.2 μm vent caps (i.e., filtering caps) made of PTFE (polytetrafluoroethylene) for enhanced aeration, which can provide optimal growth of selected cell lines. Another preferred labware for maintaining cell cultures is the bioreactor tube, which is a sterile, threaded tube with 0.2 μm PTFE vent caps (i.e., filtering caps).

[0036] In one embodiment, a holder for a flask, such as a 125 ml Erlenmeyer flask (1), is provided. The flask holder is referred to herein as a basket. The terms "(shaker) flask holder" and "(shaker) flask basket" may be used synonymously. In particular, the cell culture labware may be a flask and the cell culture labware holder (basket) is a flask holder (basket). Since the flask holder is placed in an orbital shaker incubator, it is referred to herein as a shaker flask holder. In one embodiment, the flasks maintain a CHO-M (Chinese Hamster Ovary) seed train cell line (Selexis). In one embodiment, a flask holder is provided that can have outer dimensions of 87.26±0.07 mm width and 85.64±0.08 mm length. Use of a flask holder of this dimension is compatible with other known automated laboratory equipment that is compatible with MTP labware, such as the MTP deck of the VANTGE 2.0 liquid handling system (Hamilton). Thus, the flask holder described herein is consistent with the dimensions defined by the Society for Biomolecular Screening (SBS) for standard plates.

[0037] In one embodiment, a flask holder (2) is provided that includes at least a base (2.1), two handles (2.2), a heat transfer pad (2.3), a locking means (e.g., a combination of 2.4, 2.5, 2.6, 2.7), and may further include a counter magnetic pad (2.8) and an identification means (2.9).

[0038] Possible elements of the flask holder can be described as follows: -Bottom(2.1) In one embodiment, the flask holder has a bottom surface that carries two handles (2.2), a heat transfer pad (2.3), a tooth spring with a horseshoe shape (2.4), a safety latch (2.5), a magnetic latch (2.6), a latch embossment (2.7), a counter magnetic pad (2.8), and further elements of the holder such as an RFID tag or QR code (2.9). In one embodiment, the bottom surface is as seen in Figures 1A-E. In one embodiment, the bottom surface is preferably made from PEEK polymer (polyetheretherketone polymer). In general, the use of PEEK polymer prevents corrosion inside the culture shaker compartment.

[0039] -Handle(2.2) In one embodiment, one or two handles are disposed on the flask holder bottom surface. Preferably, two handles are disposed on the flask holder bottom surface such that the handles are perpendicular to the bottom surface and face each other. In one embodiment, the handle is rectangular in shape, preferably hollow, and at least one side is molded to provide a gripping side that fits with a gripping means such as a three-axis motorized stage or a four-axis collaborative robot. In another embodiment, the handle is rectangular in shape, preferably hollow, and at least one side is embossed to provide a gripping side that fits with a gripping means such as a three-axis motorized stage or a four-axis collaborative robot. The shape of the handle allows for gripping by a gripping means (e.g., a gripping means of a three-axis motorized stage (4.7) or a gripping means of a four-axis collaborative robot) and may be referred to herein as a gripping handle. The handle may be attached to the bottom surface by any attachment means, such as screws. In one embodiment, the handle is preferably made of hard anodized aluminum alloy. In general, the use of hard anodized aluminum alloy prevents corrosion inside the culture shaker compartment. In one embodiment, the two handles can be arranged as seen in Figures 1A-E. In one embodiment, the height of the handle may be modified to accommodate and still allow for gripping of labware of different heights.

[0040] -Heat Conduction Pad (2.3) In one embodiment, the thermally conductive pad fits within a hollow cylindrical opening on the bottom surface of the flask holder and is positioned below the bottom surface of the flask. In one embodiment, the thermally conductive pad (2.3) is as seen in Figure IE. Another embodiment is shown in Figures 9A and 9B. The heat transfer pad is positioned to provide direct heating to a flask placed in the flask holder. A heat transfer pad is an example of a means for providing heat to a flask placed in the flask holder. The heat transfer pad is made from a heat conducting material, preferably a material with high thermal conductivity such as metal, that conducts heat. Heat transfer occurs when the material is placed on a heated surface, whereby the material favors heat exchange with, for example, a flask. In one embodiment, the thermally conductive pad is preferably made from a hard anodized aluminum alloy. In the example shown in Figure 1E, a heat transfer pad 2.3 is placed at the bottom of the flask holder and designed to contact the flat surface of the flask bottom. Heat is transferred from the tray to the flask via the heat transfer pad 2.3. In another embodiment, the thermal conduction pads (10.9) extend along the sides of the flask. In the example shown in Figure 9A, four thermal side conductors (10.8) are connected to the bottom pad (10.9) to provide heat along the sides of the flask. Note that the side heat conductor (10.8) can be used for a basket with one Erlenmeyer flask or for a basket with four tube flasks as shown in FIG.

[0041] - Locking means (for flasks - 2.4, 2.5, 2.6, 2.7 or for bioreactor tube combinations - 10.1, 10.2, 10.3, 10.4, 10.5, 10.6, 10.7) In one embodiment, the locking means is located on the bottom surface of the flask holder. In an alternative embodiment, the locking means is suitable for Erlenmeyer flasks and may include at least one of a toothed clamping means (e.g. 2.4) with a latch embossment (e.g. 2.7) and a latch (e.g. safety latch (2.5)) with a magnetic latch (e.g. 2.6). The function of this locking means is to keep the flask from rotation and vertical movement. This locking means for Erlenmeyer flasks is located on the flask holder bottom surface as seen in Figure 1. In another embodiment, the locking means is suitable for bioreactor tube flasks and comprises four guillotines (10.1) each holding a toothed jaw (10.2), and is placed on the bottom surface of the cell culture flask holder. In another embodiment, the locking means is suitable for bioreactor tube flasks and may be a combination of four guillotines (10.1), eight toothed jaws (10.2), four rails and pivots (10.3) (pivots not visible), plunger (10.4), plunger holder (10.5), eccentric jig (10.6) and control latch (10.7). The function of this locking means is to keep up to four bioreactor tubes from rotation and vertical movement. This locking means for the bioreactor tubes is located on the bottom surface of the flask holder as seen in Figure 9.

[0042] Locking means for Erlenmeyer flasks In one embodiment, the locking means for the Erlenmeyer flask is located on the bottom surface of the holder and perpendicular to the bottom surface. The locking means is located on the bottom portion of the holder where the handle is not attached. The locking means may be attached to the bottom surface by any attachment means (e.g. a pivot).

[0043] Clamping means (2.4) In one embodiment, a clamping means (or element) for the bottom surface of the flask fits into a hollow cylindrical opening on the bottom surface of the flask holder and is positioned to maintain the bottom surface of the flask. The function of this clamping element is to keep the flask from rotation and vertical movement. The clamping element can be attached to the bottom surface by any attachment means, such as a pivot. In one embodiment, the flask is an Erlenmeyer flask and the clamping element is a circular toothed spring with a horseshoe shape. The function of this element is to keep the Erlenmeyer flask from rotation and vertical movement. The circular spring is arranged so that when the clamping element is placed around the Erlenmeyer flask bottom, it contacts (latches) at least 60% of the cylindrical Erlenmeyer flask bottom. The circular spring has additional toothed embossings on parts of the edges, which contribute to maintaining the position of the Erlenmeyer flask. In one example, a toothed spring (2.4) with a horseshoe shape for holding the Erlenmeyer flask can be seen as in Figure 1B-D. The circular spring has additional embossments at one or two spring ends, referred to herein as latch embossments (2.7). In one embodiment, the toothed clamping means has at least one, preferably at least two latch embossments.

[0044] The latch embossment is designed to latch the safety latch (2.5). The latch embossment contributes to the clamping of the flask. In one embodiment, the latch embossment (2.7) that latches the safety latch (2.5) can be seen as in FIG. 1C. ·Safety latch(2.5) In one embodiment, the locking means is exemplified by a latch further comprising at least one magnet inserted in the latch and by at least one opposing magnet arranged on the bottom surface of the holder. The inserted at least one magnet can be glued to the latch and the at least one opposing magnet can be glued to the bottom surface of the holder. The at least one inserted magnet can be a permanent magnet. In one embodiment, at least two permanent magnets are used and inserted in the latch. In one embodiment, at least two opposing magnets are used and placed on the bottom surface of the flask holder, whereby the magnets are perpendicular to the bottom surface. In another exemplary embodiment, two permanent magnets inserted in the latch and two opposing magnets placed on the bottom surface can be used, as seen in FIG. 1C. In one embodiment, the magnet pair together is referred to as a magnetic latch (2.6). It is understood that the at least one inserted magnet in the latch and the at least one opposing magnet placed on the bottom surface are attached by magnetic attraction to each other when the safety latch is in the closed position, as seen in FIG. 1B. Thus, the magnet on the latch and a set of opposing magnets on the bottom surface provide additional latching means to secure the toothed clamping means in the closed position, as seen in Figure 1B. This latch may be referred to herein as a safety latch (2.5). The at least one magnet inserted into the latch and at least one counter magnet disposed on the bottom surface of the holder are referred to herein as a magnetic latch (2.6) (FIGS. 1D-E). In an embodiment, the safety latch is made from a hard anodized aluminum alloy. In one embodiment, the clamping means is moved away from the holder bottom surface so that the flask is placed on the bottom surface of the holder. The clamping means is then lowered and placed around the flask bottom surface. Finally, the safety latch is lifted, allowing at least one interposed magnet in the latch and at least one counter magnet on the bottom surface to attach to each other, and then the safety latch is secured with the latch embossment of the clamping means. In one example, the latch embossment (2.7) of the clamping means (2.4) may latch the safety latch (2.5) when the clamping means and the safety latch are in the closed position as seen in FIG. 1B.

[0045] · Locking means for bioreactor tube flasks (10) In one embodiment, the locking means for the bioreactor tube (11) is located on the bottom surface of the holder and is perpendicular to the bottom surface. The locking means can be attached to the bottom surface by any attachment means (e.g. screws). In one embodiment, the locking means for up to four bioreactor tubes (11) may include four guillotines (10.1), eight toothed jaws (10.2), four rails and pivots (10.3) (pivots not visible), a plunger (10.4), a plunger holder (10.5), an eccentric jig (10.6), and a control latch (10.7). In one embodiment, the locking means for the bioreactor tube (11) comprises four guillotines (10.1) that hold the toothed jaws (10.2). The four guillotines are driven by four rails and pivots (10.3) located in the plunger (10.4). The up and down movement of the plunger (10.4) is driven by an eccentric jig (10.6). The construction (10) is connected to a control latch (10.7). It is understood that up to four bioreactor tubes can fit into one flask holder. Although exemplary alternative locking means for the Erlenmeyer flask and bioreactor tube are used, it is understood that this does not affect the technical specifications of the other flask holder elements and their function. Similarly, the exemplary alternative locking means used in the flask holder does not affect the way the flask holder is handled. The flask holder with locking means for the bioreactor tube is referred to herein as a flask basket for the bioreactor tube (12).

[0046] - Opposing magnetic pads (2.8 and 10.10) In one embodiment, at least one, at least two, at least three, at least four opposing magnets are disposed on the exterior of the flask holder bottom surface. Preferably, four opposing magnets are disposed on the exterior of the flask holder bottom surface. In one embodiment, the opposing magnets are glued to the exterior of the flask holder bottom surface. In one embodiment, the opposing magnets are shaped as magnetic pads (2.8 and 10.10), as seen in Figures IE and 9A. In an embodiment, the counter magnetic pad is made of magnetizable stainless steel (ferritic or martensitic stainless steel). In general, the use of stainless steel prevents corrosion inside the culture shaker compartment. The horizontal position of the flask holder on the shaker tray is ensured by at least two magnets that can be attached by magnetic attraction to at least two conical positioning guide members present on the shaker tray and at least two opposing magnetic pads located on the outside of the bottom surface of the flask holder present on the shaker tray. The horizontal position of the flask holder on the flask holder plate of the transport shuttle is ensured by up to four conical positioning guide members present on the flask holder plate. The function of these opposing magnetic pads is to further maintain the flask holder in an upright position.

[0047] -Means of Identification (2.9) In one embodiment, the flask holder is configured with an identification means, such as RFID (radio frequency identification) or a two-dimensional barcode (e.g., QR code), to allow for culture information tracking (cell line ID, passage number, passaging date, etc.). In one embodiment, the flask holder has an RFID tag or QR code (2.9). In one embodiment, the RFID tag or QR code (2.9) is placed on the outside of the handle, opposite the gripping side, as seen in FIG. 1E with the QR code (alternatively, the RFID tag can be placed under the bottom of the flask holder). The RFID is obtained through a reader / writer communication with the culture shaker's PLC (Programmable Logic Circuit), which then uses this RFID information to generate a database for tracking the baskets while in use, which can be used as a safety net in case of failure of scheduling software such as the Hamilton scheduling software. In one embodiment, the size of the flask holder is adjusted to accommodate all the flasks. In one embodiment, the flask placed in the holder is an Erlenmeyer flask, and the volume of the flask can be selected from about 25 ml to 4000 ml, such as 25 ml, 50 ml, 125 ml, 250 ml, 500 ml, 1000 ml, 2000 ml, or 4000 ml. In one embodiment, the flask is an Erlenmeyer flask, and the bottom diameter of the flask can be selected from about 4 cm to 20 cm, such as 4.1 cm, 5.1 cm, 6.7 cm, 8.2 cm, 10.1 cm, 12.9 cm, 16 cm, or 20.6 cm. In one embodiment, the flask is an Erlenmeyer flask, and the height of the flask can be selected from about 6 cm to 36 cm, such as 6.1 cm, 7.8 cm, 11.4 cm, 13.2 cm, 17.6 cm, 21.6 cm, 26.8 cm, or 36.0 cm. In one embodiment, the flask is a 125 ml Erlenmeyer flask. In one embodiment, the flask is a 125 mL Erlenmeyer flask (Thomson Optimum Growth™ 125 mL Flask; reference number 931110). In one embodiment, the flask placed in the holder is a bioreactor tube and the volume of the bioreactor tube may be selected from about 15 ml to 600 ml, for example, 15 ml, 50 ml, 450 ml, or 600 ml. In one embodiment, the bioreactor tube may have a conical or circular bottom. In one embodiment, the flask is a bioreactor tube and the dimensions may be selected from 17.1x120 mm, 30x115 mm, 96x145 mm, and 98x183 mm. In one embodiment, the flask is a bioreactor tube (TPP TubeSpin® Bioreactor 15), which may be conical bottom, allowing for a smaller working volume of 1-15 mL. In one embodiment, the flask has a closure cap (1.1; 11.1) that has an upper end and adjoins the side wall of the flask along the periphery of the flask neck. The closure cap is removably connected to the flask. Good sealing of the interior space of the flask against the surrounding atmosphere and also a secure fixing of the closure cap to the flask is achieved in an advantageous embodiment of the invention by a closure cap configured as a screw-on closure. The closure cap may be a screw-on cap (1.1; 11.1), which may be a filtering cap. Such a cap has the advantage of providing increased aeration while maintaining sterile conditions for the cell culture.

[0048] Culture shaker tray In one embodiment, an incubator tray (3) is provided for holding at least one flask holder (2, 12) as described herein. The incubator tray (3) is suitable for use inside an orbital shaking incubator and may be referred to herein as a culture shaker tray. In one embodiment, the incubator tray can accommodate up to 40 flask holders, such as 40 flask holders for 125 ml Erlenmeyer flasks arranged in an 8x5 array as seen in Figure 2 C. The function of the incubator tray is to maintain the flask holders in a stable position while the orbital shaking incubator is running. In one embodiment, the shaker tray is preferably made from a hard anodized aluminum alloy, which prevents corrosion inside the culture shaker compartment of the tray. In one embodiment, an incubator tray (3) is provided that comprises a lower plate (3.1), an upper plate (3.2) and a tray locker (3.3).

[0049] Possible elements of the incubator can be described as follows: -Lower plate (3.1) In one embodiment, the lower plate (3.1) is placed below the upper plate (3.2) and is attached to the upper plate (3.2) by any attachment means, preferably by screws. In one embodiment, the lower plate is preferably made from a hard anodized aluminum alloy. In one embodiment, the lower plate is about 3 mm to about 5 mm thick, preferably about 4 mm thick. In one embodiment, the lower plate (3.1) has up to 80 electromagnets located on the top of the lower plate. The electromagnets can be screwed to the top of the lower plate. In one embodiment, permanent electromagnets (3.4) can be used. A permanent electromagnet (3.4) is induced in the top plate (3.2) through a hole. The magnetic field of the permanent electromagnet is suppressed when the coil is activated. The permanent electromagnet is divided into eight parts in the tray, each one controlled by a solid state relay. The relays are controlled via a controller means such as an I2C bus by a microcontroller (e.g. an Arduino controller) which communicates to a Programmable Logic Controller (PLC) (e.g. a Festo) via a ME3 / ME4 control card (e.g. a Coppley control). In one embodiment, the lower plate (3.1) has up to 40 RFID readers / writers located on the top of the lower plate. The RFID readers / writers communicate to a microcontroller (e.g., Arduino) via I2C protocol. The RFID readers / writers may include built-in antennas. In one embodiment, the lower plate is as shown in Figure 1 A. In one embodiment, the lower plate constructed with the upper plate is as shown in Figure 1B.

[0050] -Upper plate (3.2) In one embodiment, the upper plate (3.2) is placed on the lower plate (3.1) and attached to the upper plate (3.1) by any attachment means, preferably by screws, and maintained together at a certain distance via spacers. In one embodiment, the distance between the lower plate (3.1) and the upper plate (3.2) placed above is about 10 mm to about 11 mm, preferably about 10.5 mm. In one embodiment, the upper plate is preferably made from a hard anodized aluminum alloy. In one embodiment, the top plate is about 3 mm to about 5 mm thick, preferably about 3 mm thick. In one embodiment, the top plate has at least one guide member disposed on the top of the top plate. In one embodiment, the top plate has up to 80 guide members disposed on the top of the top plate. The guide members may be attached to the top of the top plate by any attachment means, preferably by screws. The guide members are referred to herein as centering guide members or positioning guide members. The guide members may be positioned such that two guide members are positioned for each flask basket position. The centering guide members may have a cone shape. The centering guide members may be exemplified by pins, such as positioning pins (3.5). In one embodiment, the flask holder is placed between two guide members present on the incubator tray, which provide a horizontal position for the flask holder. In one embodiment, the top plate (3.2) has up to 40 holes for RFID reader / writer antennas, referred to herein as RFID antenna holes (3.6). It is understood that the opposing magnets on the flask holder (2.8) and the electromagnets (3.4) on the tray coincide via the guide features of guide members (e.g., locating pins (3.5)) located on the upper plate (3.2).

[0051] -Tray Locker (3.3) In one embodiment, the tray locker (3.3) is placed on the track tray holder. In one embodiment, the tray locker (3.3) provides precise positioning of the trays in two ways: first, by two half-moon millings on the tray that correspond to two cylindrical jigs on the back side of the tray, and second, by fixing the tray in a precise position fixed by an eccentric handle on the front side of the tray. In one embodiment, the tray locker (3.3) is compatible with automation as described herein.

[0052] Three-axis motorized stage In one embodiment, a three-axis motorized stage (4) is provided that is designed to be mounted inside the top wall (ceiling) of the orbital culture shaker chamber. The three-axis motorized stage is attached to the orbital culture shaker chamber by any attachment means, preferably by stainless steel threaded dowels and non-thermally conductive PEEK nuts. It will be appreciated that the three-axis motorized stage can be a compact system (with two-stage extension) that increases the operable height of the gripping system. Such a compact (low height) three-axis motorized stage allows for the handling and transport of tall flask holders, such as flask holders for Erlenmeyer flasks or bioreactor tubes. In one embodiment, the load capacity of the three-axis motorized stage is 0.8 kg. In one embodiment, a three-axis motorized stage (4) is provided, including an X-axis (4.1), a Y-axis (4.2), a Z-axis (4.3), and at least one motor for driving each axis. In one embodiment, the motors for driving the X-axis or the Y-axis or the Z-axis (4.3) are linear motors. In one embodiment, at least one encoder is provided, providing feedback for closed-loop control of the at least one motor for driving each axis. In one embodiment, as seen in FIG. 3, a three-axis motorized stage (4) is provided, comprising an X-axis (4.1), a Y-axis (4.2), a Z-axis (4.3), a linear motor (4.4) for driving the X-axis (4.1), a linear motor (4.5) for driving the Y-axis (4.2), a linear motor (4.6) for driving the Z-axis (4.3), and a means for gripping a flask holder (4.7).

[0053] Possible elements of a three-axis motorized stage can be described as follows: -X axis (4.1) In one embodiment, the X-axis has a range of travel of about 685 mm. Preferably, the three-axis motorized stage has one X-axis. In one embodiment, the X-axis (4.1) is connected to one linear motor (4.4). -Y axis (4.2) In one embodiment, the Y-axis has a range of motion of about 375 mm. Preferably, the three-axis motorized stage has one Y-axis. -Z axis (4.3) In one embodiment, the Z axis has a range of motion of about 175 mm. Preferably, the three-axis motorized stage has one Z axis.

[0054] - A linear motor (4.4) for driving the X-axis (4.1) In one embodiment, the X-axis (4.1) is connected to one linear motor (4.4), such that the linear motor drives the X-axis. In one embodiment, the linear motor driving the X-axis is selected from linear motors known in the art, such as a TM6 linear motor (Technotion). In one embodiment, an LA11 absolute encoder (RLS) is provided, providing feedback for closed loop control of the linear motor (4.4) via the ME3 card.

[0055] - Linear motor (4.5) for driving the Y-axis (4.2) In one embodiment, the Y-axis (4.1) is connected to one linear motor (4.5), such that the linear motor drives the Y-axis. In one embodiment, the linear motor driving the Y-axis is selected from linear motors known in the art, such as a TM6 linear motor (Technotion). In one embodiment, an LA11 absolute encoder (RLS) is provided, providing feedback for closed loop control of the linear motor (4.5) via the ME3 card.

[0056] - A linear motor (4.6) for driving the Z-axis (4.3) In one embodiment, the Z-axis (4.3) is connected to one linear motor (4.6), such that the linear motor drives the Z-axis. In one embodiment, the linear motor driving the Z-axis is selected from linear motors known in the art, such as a TM3 linear motor (Technotion). The Z-axis provides the extension motion. In one embodiment, an LA11 absolute encoder (RLS) is provided, providing feedback for closed loop control of the linear motor (4.6) via the ME3 card. In one embodiment, the linear motor may be disposed within the case (as seen in FIG. 3).

[0057] -means for gripping the flask holder In one embodiment, means for gripping the flask holder are provided. The means for gripping the flask holder are attached to the Z axis (4.3) by any attachment means such as a screw. The function of these gripping means is to allow gripping of the flask holder for transport. The function of the gripping means is to allow a detachable connection to the flask holder. The gripping means must therefore be compatible with the flask holder handle. In one embodiment, an example of a means for gripping the flask holder is one parallel gripper (4.7) with adjustable gripping force, with jaws (4.8) and fingers (4.9) for removable connection to the flask holder. In one embodiment, the parallel gripper is selected from parallel grippers known in the art, such as the motorized EHPS parallel gripper (Festo). In an embodiment, the parallel grippers are made from a hard anodized aluminum alloy. In an embodiment, the jaws of the parallel gripper are made from stainless steel. In an embodiment, the parallel gripper fingers are made from a hard anodized aluminum alloy. In an embodiment, the parallel gripper is controlled by a PLC, which communicates via an IO-Link interface.

[0058] Sliding door In one embodiment, a sliding door (5) is provided that is designed to be mounted within a side wall of an orbital culture shaker chamber. The sliding door (5) is designed to be mounted within a rounded rectangular hole drilled on the side wall of the orbital culture shaker chamber. Preferably, the sliding door (5) is designed to be mounted on the left side wall of the orbital culture shaker chamber. In one embodiment, the sliding door is automated. In one embodiment, the sliding door is attached to the orbital culture shaker chamber by any attachment means, preferably by screws. In one embodiment, the sliding door includes a door frame (5.1) and a door panel (5.2), which forms a door tunnel (5.3) when the door panel is open. In another further embodiment, the door panel (5.2) is composed of three parts: a front panel (5.2.1) (Figure 4B), a main body (5.2.2) (Figure 4C) and a pressure panel (5.2.3) (Figure 4D). In one embodiment, a sliding door (5) is provided, comprising a door frame (5.1) and a door panel (5.2) and at least one motor for driving the movement of the door. In one embodiment, a sliding door (5) is provided, comprising a door frame (5.1) and a door panel (5.2) and a sliding movement driving stepper motor (5.4) and a pressure movement driving stepper motor (5.5). The sliding door may further comprise at least one sensor for the movement of the door. The sliding door may further include heating means such as a temperature probe (5.8), a heating pad (5.9), a heating ribbon (5.11) and a control device. The sliding door may further include a precision sensor (5.6) for the sliding movement and a precision sensor (5.7) for the pressure movement, a temperature probe (5.8), a heating pad (5.9), an O-ring (5.10) and a heating ribbon (5.11) as seen in Figure 4.

[0059] Possible elements of a sliding door can be described as follows: -Door Frame (5.1) -Door panel (5.2) The door panel of a door slides open and closed. When the door panel is open, it forms a door tunnel (5.3), which is the space through which goods travel when the door is open. The door panel (5.2) is composed of three parts: a front panel (5.2.1) (Figure 4B), a body (5.2.2) (Figure 4C) and a pressure panel (5.2.3) (Figure 4D). In another embodiment, the front panel (5.2.1) is arranged with a sliding motion driving stepper motor (5.4) and a pressure motion driving stepper motor (5.5). In one embodiment, the body (5.2.2) is arranged between the front panel (5.2.1) and the pressure panel (5.2.3). In one embodiment, the pressure panel (5.2.3) is arranged with a temperature probe (5.8) and a heating pad (5.9), preferably said temperature probe and said heating pad are embedded in the pressure panel. In one embodiment, when the door is closed, the pressure panel (5.2.3) is in contact with an O-ring (5.10). The sliding door exerts pressure against the O-ring.

[0060] - Two stepper motors: a sliding motion drive stepper motor (5.4) and a pressure motion drive stepper motor (5.5) In one embodiment, the sliding door includes at least one motor for driving the door movement, such as a sliding movement driving stepper motor (5.4) and a pressure movement driving stepper motor (5.5). In one embodiment, the stepper motor is selected from stepper motors known in the art, such as NEMA-23. In one embodiment, the stepper motor is controlled by a controller means, such as an ME4 control card. In one embodiment, one stepper motor drives the sliding motion (5.4) via a simple rack and pinion. In one embodiment, one stepper motor driving the sliding motion is located on the right side of the door, such as on the front panel (5.2.1). In one embodiment, one stepper motor drives the compression movement (5.5) via a motor gearbox, two toothed wheels and four satellite wheels. The four satellite wheels drive four lead screws that control the compression movement. Two sliding ball bearings guide the whole mechanism. In one embodiment, one stepper motor that drives the sliding motion is located on the left side of the door, such as on the front panel (5.2.1).

[0061] - Two sensors: a precision sensor for sliding movements (5.6) and a precision sensor for pressure movements (5.7) In one embodiment, the sliding door includes at least one sensor for the movement of the door, such as a sensor for the sliding movement (5.6) and a sensor for the pressing movement (5.7). In one embodiment, the sensor is a precision sensor. In one embodiment, the precision sensor is selected from sensors known in the art, such as a precision sensor switch (Baumer). In one embodiment, the sensor is controlled by a controller means, such as an ME4 control card, which is ultimately controlled by a PLC. In one embodiment, one precision sensor is dedicated to the detection of sliding motion when the door is opened (there is no detection when the door is closed). It is placed on the door frame (5.1). It is referred to herein as precision sensor (5.6) for automatic induction of the precision sensor of sliding motion. In one embodiment, one precision sensor is dedicated to the detection of the pressurization movement. It is placed on the sliding door panel (5.2). It is referred to herein as the precision sensor (5.7) for automatic induction of the pressurization movement when the door is not pressurized (such as an open door).

[0062] -Temperature probe (5.8) In one embodiment, the temperature probe is selected from temperature probes known in the art (5.8), such as a Pt1000 probe. The temperature probe used is suitable for precision temperature sensing. In one embodiment, at least one temperature probe is used. It is placed on the door, such as embedded in the pressure panel (5.2.3). In one embodiment, the temperature probe is connected to a microcontroller (e.g., an Arduino controller) that controls the heating pad (5.9) and / or heating ribbon (5.11). The microcontroller communicates the door temperature value to a PLC for temperature monitoring.

[0063] -Heating pad (5.9) In one embodiment, the heating pad is selected from heating pads known in the art (5.8), such as polyimide encapsulated heating pads. In one embodiment, at least one heating pad is used. In one embodiment, one polyimide encapsulated heating pad is embedded in the door, such as embedded in the pressure panel (5.2.3). The use of a heating pad allows for door temperature control and prevents agglomeration on the door surface.

[0064] -O-ring (5.10) In one embodiment, the O-ring is selected from O-rings (5.10) known in the art, such as silicone O-rings. O-rings may also be called countour rings. In one embodiment, at least one O-ring is used. In one embodiment, one silicone O-ring is installed between the door frame (5.1) and the pressure panel (5.2.3) of the door panel (5.2). The use of an O-ring ensures an airtight seal.

[0065] -Heating ribbon (5.11) In one embodiment, the door frame (5.1) is arranged with at least one heating ribbon (5.11) disposed around the opening forming the door tunnel (5.3). In one embodiment, the heating ribbon is selected from heating ribbons known in the art, such as polyimide encapsulated heating ribbons. In one embodiment, at least one heating ribbon is used. The use of a heating ribbon allows for tunnel temperature control and prevents agglomeration on the tunnel surface.

[0066] Transport Shuttle In one embodiment, a transport shuttle (6) is provided that is designed to be mounted on the outside of the side wall of the orbital culture shaker chamber. Preferably, the transport shuttle (6) is designed to be mounted on the left side wall of the orbital culture shaker chamber. In one embodiment, the transport shuttle is automated. In one embodiment, the transport shuttle is attached to the orbital culture shaker chamber by any attachment means, preferably by screws. The transport shuttle (6), for example shown in Figures 6A-6C, is designed to be attached to the outside of the sidewall of the orbital culture shaker chamber at the bottom of the sliding door (5), as seen in Figure 6. This allows movement of the flask holder (6.5) through the sliding door tunnel (5.3) that is formed when the sliding door (5.2) is open. In one embodiment, the automated transport shuttle (6) may include at least a slide frame (e.g. 6.1), rails (e.g. 6.2), at least two belts (e.g. 6.11) disposed on the slide frame, a cell culture flask holder plate (e.g. 6.5) and a motor (e.g. 6.3) for driving the sliding movement of the plate on the rails. In one embodiment, the automated transport shuttle (6) may include at least a sliding frame (6.1) and rails (6.2), a sliding motion drive stepper motor (6.3), a ball screw (6.13), a flask holder plate (6.5), a belt (6.11), a flask holder plate (6.5) having two guide members (6.6). The automated transport shuttle (6) may further include a pinion (6.4), an amplified precision sensor (6.7) for detecting the transport shuttle home position, a sensor (6.8) for detecting the flask holder attachment position and a sensor (6.9) for detecting the flask holder removal position, and a presence laser detector (6.10) as seen in Figures 5 or 6.

[0067] Possible elements of the transport shuttle (6) can be described as follows: - Slide frame (6.1) and rail (6.2) and sliding motion drive motor (6.3) In one embodiment, at least one frame is used, hi one embodiment, the frame is made of stainless steel. In one embodiment, the sliding frame (6.1) holds at least two rails. In another exemplary embodiment, the sliding frame (6.1) holds four rails (6.2), two on the top and two on the bottom. The top plate can slide on the top rails. The sliding frame slides on the shuttle base through the bottom rails. In one embodiment, the sliding motion drive motor is a stepper motor (6.3). The sliding is controlled by a stepper motor (6.3) via a ball screw (6.13) (first motion) (FIG. 5). In one embodiment, the stepper motor is selected from stepper motors known in the art, such as a NEMA-23. In one embodiment, a stainless steel frame slides on the rails and the sliding motion is controlled by a NEMA-23 stepper motor via a ball screw (6.13) (first motion). In one embodiment, a stepper motor driving the sliding motion is placed below the rails. In one embodiment, the stepper motor is controlled by a controller means, such as an ME4 control card, which is ultimately controlled by a PLC.

[0068] -Pinion (6.4) In one embodiment, the pinion (6.4) is mounted on the slide frame (6.1) with a pinion holder screwed onto it. In one embodiment, the pinion (6.4) drives the two-stage extension of the upper plate (6.12) via a rack (second motion).

[0069] -Flask holder plate (6.5) In one embodiment, at least one flask holder plate is used. The flask holder slides on a rail (6.2). The sliding movement of the flask holder (6.5) on the rail (6.2) is driven by at least two belts (6.11) arranged on the slide frame (6.1) (third movement). In another embodiment, the sliding movement of the flask holder (6.5) on the rail (6.2) is driven by at least four belts (6.11) arranged on the slide frame (6.1) (third movement). In one embodiment, the belts are glued and screwed on one side on the slide frame and on the other side on the flask holder. A tensioning device is installed on one side. A pulley installed on a two-stage tension plate (6.12) reverses the direction of movement. The belts are preferably made of polyimide film. In general, the use of polyimide film provides higher mechanical resistance (does not stretch) and because it is thinner than other common materials used in belts, it allows for more compact belts while still maintaining their resistance.

[0070] In one embodiment, the flask holder plate (6.5) comprises at least two guiding elements attached to the top of the flask holder plate by any attachment means, preferably by screws. The guiding elements are referred to herein as centering or positioning guiding elements. Preferably, four centering guiding elements are arranged for the flask holder. The guiding elements may have a conical shape. The centering guiding elements may be exemplified by pins, such as positioning pins (6.6). In one embodiment, the flask holder is placed between at least two or at least four guiding elements (e.g. pins) present on the flask holder plate, which provide a horizontal position for the flask holder.

[0071] - Position sensors / detectors including an amplified precision sensor (6.7) for detecting the transport shuttle home position, a sensor (6.8) for detecting the flask holder attachment position, and a precision sensor (6.9) for detecting the flask holder removal position. In one embodiment, the transport shuttle includes at least one sensor for detecting a home position of the transport shuttle, such as a home position of the flask holder and / or an attachment position and / or an detachment position of the flask holder, and thus preferably referred to herein as a position sensor / detector. In one embodiment, the sensor is selected from an amplified precision sensor or a precision sensor.

[0072] · Amplified precision sensor for measuring transportation shuttle home position (6.7) In one embodiment, the sensor (6.7) for detecting the transport shuttle home position can be an amplified precision sensor. In one embodiment, the amplified precision sensor is selected from sensors known in the art, such as a precision sensor switch (Baumer). In one embodiment, the sensor is controlled by a controller means, such as an ME4 control card, which is in turn controlled by a PLC. In one embodiment, preferably at least one amplified precision sensor is dedicated to detecting the home position of the transport shuttle. It is placed on the shuttle frame and detects the position of the slide frame. It is referred to herein as precision sensor for detecting transport shuttle home position (6.7). The home position is shown in Figure 5 or 6 and represents the idle state, defined as the non-extended state where the flask holder plate is located outside the incubator compartment.

[0073] Two precision sensors (6.8) and (6.9) for detection of the flask holder attachment / detachment position In one embodiment, the sensors (6.8) and (6.9) for detection of the attachment / detachment position of the flask holder can be precision sensors. In one embodiment, the amplified precision sensor is selected from sensors known in the art, such as a precision sensor switch (Metrol). In one embodiment, the sensor is controlled by a controller means, such as an ME4 control card, which is in turn controlled by a PLC. In one embodiment, preferably at least one precision sensor is dedicated to the detection of the flask holder attachment position, i.e. when the flask holder is placed on the flask holder plate inside the culture shaker compartment. In one embodiment, preferably at least another precision sensor is dedicated to the detection of the flask holder removal position, i.e. when the flask holder is removed from the flask holder plate outside the culture shaker compartment. The two sensors are referred to herein as sensor (6.8) for the detection of the attachment position of the flask holder and sensor (6.9) for the detection of the removal position of the flask holder. The two sensors for the detection of the attachment / removal position of the flask holder are arranged on opposite sides of the two-stage extension plate (6.12). The attachment position represents the state when the transport shuttle is extended and the flask holder plate is located inside the culture shaker compartment. The removal position corresponds to the home position shown in Figure 5 or 6 and is defined as the non-extended state where the flask holder plate is located outside the culture shaker compartment.

[0074] Presence Laser Detector (6.10) The laser detector makes it possible to detect the presence or absence of a flask holder (basket) and is therefore preferably referred to herein as a presence laser detector. In one embodiment, the presence laser detector is selected from the presence laser detectors known in the art. The presence laser detector preferably used is suitable for measuring distances, such as the distance between a basket on a shuttle inside the incubator and a basket outside the incubator. If a flask holder is present, the presence laser detector makes it possible to identify whether the flask holder can be grasped inside / outside the incubator. In one embodiment, at least one presence laser detector is used. The presence laser detector may be located on the outer end of one of the transport shuttles. The presence laser detector may be directly controlled by a control device, such as by a PLC, communicating via an IO-Link interface.

[0075] Automated Culture Shaker Compartment In one embodiment, - culture shaker trays (3), - Three-axis motorized stage (4), -sliding door (5), and -Transport shuttle (6), A construct designed to be placed in an orbital culture shaker chamber is provided, comprising: a set of at least one cell culture flask disposed in at least one cell culture flask holder; this set of elements is essential for maintaining and subsequently transporting at least one cell culture flask disposed in at least one cell culture flask holder; and is therefore referred to herein as a set, or alternatively, as an apparatus. In one embodiment, the culture shaker tray functions to maintain at least one cell culture flask holder. In one embodiment, the function of the three-axis motorized stage is to initially maintain and subsequently simultaneously maintain and transport one cell culture flask holder. In one embodiment, the function of the sliding door is to enable transport of one cell culture flask holder inside / outside the orbital culture shaker chamber. In one embodiment, the function of the transport shuttle is to initially maintain and subsequently simultaneously maintain and transport one cell culture flask holder.

[0076] In one embodiment, - culture shaker trays (3), - Three-axis motorized stage (4), -sliding door (5), and -Transport shuttle (6), A system for an orbital culture shaker chamber is provided, comprising: In one embodiment, the culture shaker tray (3), three-axis motorized stage (4), sliding door (5) or transport shuttle (6) described herein may be used individually and may be combined with any other compatible device.

[0077] In one embodiment, a set (7) for maintaining and transporting at least one cell culture flask is provided, the set being suitable for a culture shaker compartment, and - culture shaker trays (3), - Three-axis motorized stage (4), -sliding door (5), and -Transport shuttle (6), wherein at least one cell culture flask is placed in at least one cell culture flask holder (2, 12) that is compatible with the set (7). In one embodiment, the set is an automated set. In one embodiment, the set (7) for maintaining and transporting suitable cell culture flasks to a culture shaker compartment can be referred to as an automated culture shaker compartment.

[0078] In one embodiment, the set (7) is designed to be attached to an orbital culture shaker chamber, thereby - a culture shaker tray (3) designed to be mounted inside the orbital culture shaker chamber; - a three-axis motorized stage (4) designed to be mounted inside the top wall of the orbital culture shaker chamber; - a sliding door (5) designed to fit into a rounded rectangular hole on the side wall of the orbital culture shaker chamber; - The transport shuttle (6) is designed to be attached to the outside of the side wall of the orbital culture shaker chamber. An exemplary design of the above set (7) for an orbital culture shaker chamber is shown in FIG.

[0079] In one embodiment, - culture shaker trays (3), - Three-axis motorized stage (4), -sliding door (5), and -Transport shuttle (6), The parts of set (7) of the present invention, including: have at least one feature or any combination of features described herein. In one embodiment, at least one cell culture flask holder (2, 12) compatible with the set (7) has at least one feature or any combination of features described herein. In one embodiment, the set of the present invention is suitable for attachment to any culture shaker, particularly a culture shaker having dimensions selected from a length (L) of about 1288 mm, a depth (D) of about 859-917 mm, and a height (H) of about 771 mm. In one embodiment, the set of the present invention is mounted on a culture shaker having a chamber with dimensions selected from 1288x859x771mm (LxDxH) or 1288x917x771mm (LxDxH) or 1288x887x771mm (LxDxH) or 1288x859x771mm (LxDxH), preferably 1288x859x771mm (LxDxH).

[0080] Culture shaker with automated culture shaker compartment In one embodiment, a culture shaker is provided that includes a set, device, system or assembly of elements described herein. In one embodiment, a culture shaker (8) is provided that includes a set (7) for maintaining and transporting at least one cell culture flask, the set comprising: - culture shaker trays (3), - Three-axis motorized stage (4), -sliding door (5), and -Transport shuttle (6), wherein at least one cell culture flask is placed in at least one cell culture flask holder (2, 12) that is compatible with the set (7).

[0081] In one embodiment, the set in the culture shaker is an automated set. In one embodiment, the set in the culture shaker may be referred to as an automated culture shaker. In one embodiment, a culture shaker (8) is provided that includes a set (7), the set being designed to be attached to an orbital culture shaker chamber, thereby - a culture shaker tray (3) designed to be mounted inside the orbital culture shaker chamber; - a three-axis motorized stage (4) designed to be mounted inside the top wall of the orbital culture shaker chamber; - a sliding door (5) designed to fit into a rounded rectangular hole on the side wall of the orbital culture shaker chamber; - The transport shuttle (6) is designed to be attached to the outside of the side wall of the orbital culture shaker chamber. In one embodiment, the sliding door (5) is designed to mount within a rounded rectangular hole on the side wall of the orbital culture shaker chamber.

[0082] In one embodiment, a culture shaker (8) is provided that includes a set (7), - culture shaker trays (3), - Three-axis motorized stage (4), -sliding door (5), and -Transport shuttle (6), The parts of set (7) of the present invention, including: have at least one feature or any combination of features described herein. In one embodiment, at least one cell culture flask holder (2, 12) compatible with the culture shaker (8) comprising set (7) has at least one feature or any combination of features described herein. In one embodiment, the culture shaker including the set of the present invention may be any culture shaker, particularly a culture shaker having dimensions selected from a length (L) of about 1288 mm, a depth (D) of about 859 to 917 mm, and a height (H) of about 771 mm. In one embodiment, the culture shaker comprising the set of the present invention has a chamber having dimensions selected from 1288x859x771mm (LxDxH) or 1288x917x771mm (LxDxH) or 1288x887x771mm (LxDxH) or 1288x859x771mm (LxDxH), preferably 1288x859x771mm (LxDxH). In one embodiment, a culture shaker comprising the set of the present invention is provided, wherein the culture shaker is part of an automated cell culture platform, such as an automated passaging platform. An exemplary arrangement of a culture shaker (8) including a set (7) of the present invention together with an automated cell passaging platform is shown in FIG. In one embodiment, the chamber of the liquid handling system (9) has holes that follow the contour of the automated sliding door (5) of the culture shaker (8), and the culture shaker (8) is designed such that the flask holders (2, 12) are transferred to the liquid handling system (9) with a transport shuttle (6). In one embodiment, the use of a culture shaker comprising a set of the present invention for cell culture, in particular for cell passaging, is provided. In one embodiment, a method of cell culture is provided that includes the use of a culture shaker comprising the set of the present invention.

[0083] Automated procedure for automated culture shaker It will be appreciated that the flask holders (2, 12) can be placed in and removed from the culture shaker (8) through the main front door of the culture shaker (8). Non-automated steps for handling / mounting the flask holders (2, 12) may include the following steps: a) Vacant holder position (unlock safety latch) b) placing the flask in the flask holder; c) locking the safety latch; d) opening the culture shaker through the main front door; e) Mounting the holder on the incubator tray; two pins position the holder on the tray and magnetic attraction locks the basket on the tray; f) RFID detection and verification; g) Close the culture shaker with the main front door. It will be appreciated that the flask holders (2, 12) are placed in and removed from the culture shaker (8) via a sliding door (5) of the culture shaker (8).

[0084] The automation steps for the flask holder (2, 12) transport may include the following steps: a) Place the three-axis stage at the home position. b) X / Y axis stage movement above the holder loading position, c) Z-axis stage movement to the holder loading position; d) gripping the holder by releasing the gripper fingers; e) De-energizing the holder electromagnet; f) Picking up the holder by moving the Z-axis stage; g) X / Y axis stage movement above the shuttle position, h) opening of the sliding door and extension of the shuttle towards the shaking incubator compartment (mounting position); i) Attaching the holder to the shuttle by moving the Z-axis stage; j) Return of the shuttle to the home position and re-closing of the sliding doors at the home position; k) Return of X / Y / Z stage to home position. In the case of parallel cultivation of several cultures in one orbital shaking incubator, such an automated procedure for transporting one flask holder through the set of the invention makes it possible to limit undesirable changes to the culture environment in other laboratory equipment. In one embodiment, a method is provided for maintaining at least one cell culture flask disposed in at least one cell culture flask holder using a set as described herein. In one embodiment, a method is provided for maintaining and transporting at least one cell culture flask disposed in at least one cell culture flask holder using a set as described herein.

[0085] Automated cell passaging platform including a culture shaker with an automated culture shaker compartment In one embodiment, an automated cell passaging platform is provided, comprising a liquid handling system, at least one robotized arm, at least one automated hotel, an automated refrigerated incubator, an automated microplate centrifuge with an automated loader, a cell counter and at least one shaking incubator, and optionally an automated orbital shaking incubator, characterized in that at least one orbital shaking incubator is designed to be mounted in a set to hold and transport at least one cell culture flask as described herein. In one embodiment, an automated cell passaging platform as described herein is provided, characterized in that it comprises at least two orbital shaking incubators, and the at least two orbital shaking incubators are designed to be mounted with a set for maintaining and transporting cell culture flasks as described herein. It is understood that the capacity of the automated cell passaging platform is up to 80 cell culture flasks, such as Erlenmeyer flasks, i.e. up to 40 in each orbital shaking incubator. It is understood that the capacity of the automated cell passaging platform is up to 320 bioreactor tubes, i.e. up to 160 in each orbital shaking incubator. In one embodiment, the chamber of the liquid handling system (9) has holes that follow the contour of the automated sliding door (5) of the culture shaker (8), and at least one culture shaker (8) is designed such that the flask holders (2, 12) are transferred to the liquid handling system (9) by a transport shuttle (6). In one embodiment, there is provided the use of an automated cell passaging platform comprising at least one culture shaker with an automated culture shaker compartment as described herein for cell passaging. In one embodiment, a method of cell culture is provided that includes the use of an automated cell passaging platform that includes at least one culture shaker with an automated culture shaker compartment as described herein.

[0086] References cited herein are incorporated herein by reference in their entirety. The present invention should not be limited in scope by the specific embodiments and figures described herein, which are intended as merely illustrative of individual aspects of the invention; functionally equivalent methods and elements are within the scope of the invention. The examples illustrating the present invention are not intended to limit the scope of the invention in any way.

Claims

1. A set for holding and transporting at least one cell culture flask, the set being designed to be placed in an orbital culture shaker chamber; and a flask holder, designed to receive at least one cell culture flask having a circular base area and suitable for suspension cell culture; - a culture shaker tray designed to be mounted inside the orbital culture shaker chamber, comprising at least one guiding member arranged to hold each cell culture flask holder; - an orbital culture shaker tray for receiving the flask holders and transmitting orbital motion to the flask holders; - a three-axis motorized stage designed to be mounted inside the top wall of an orbital culture shaker chamber, comprising an X-axis, a Y-axis and a Z-axis, as well as three linear motors for driving the three axes and a means for gripping a cell culture flask holder; - a sliding door designed to be mounted within a side wall of an orbital culture shaker chamber, comprising a door panel and at least one motor for driving the door movement; - a transport shuttle designed to be mounted on the outside of a side wall of said orbital culture shaker chamber, comprising a slide frame, rails, at least two belts arranged on said slide frame, a cell culture flask holder plate and a motor for driving the sliding movement of the plate on said rails; Including, set.

2. 2. The set for maintaining and transporting at least one cell culture flask according to claim 1, wherein the cell culture flask is a 125 ml Erlenmeyer flask or a bioreactor tube.

3. 3. The set of claim 1 or 2, wherein the means for gripping the cell culture flask holder comprises one parallel gripper with adjustable gripping force mounted on a Z-axis of the three-axis motorized stage and with jaws and fingers for removable connection to the flask holder.

4. 4. The set of claim 1, 2 or 3, wherein the motor for driving the door movement of the sliding door includes a stepper motor for driving the sliding movement and a stepper motor for driving the pressing movement of the door panel.

5. The set according to any one of claims 1 to 4, wherein the sliding door further comprises a precision sensor for detecting a sliding movement of the door panel and a precision sensor for detecting a pressing movement of the door panel.

6. The set of any one of claims 1 to 4, wherein the sliding door further comprises a temperature probe, a heating pad, a heating ribbon and a control device, and the temperature probe controls the heating pad and the heating ribbon via the control device.

7. The set according to any one of claims 1 to 6, wherein the sliding door further comprises a silicone O-ring mounted between the door frame and the door panel.

8. The set of any one of claims 1 to 7, wherein the transport shuttle further comprises an amplified precision sensor for detecting a home position of the transport shuttle, a sensor for detecting an attachment position of the cell culture flask holder, and a sensor for detecting a removal position of the cell culture flask holder.

9. 9. The set of any one of claims 1 to 8, wherein the flask holder comprises a bottom heat conductor on a bottom surface of the holder, one side of the heat conductor designed to contact the orbital culture shaker tray and another side of the heat conductor designed to contact the flask.

10. 10. The set of claim 9, wherein the bottom heat conductor includes four side heat conductors at four corners of the basket, which supply heat to the sides of the flask.

11. 11. The set according to any one of claims 1 to 10, further comprising a cell culture flask holder, said cell culture flask holder further comprising a bottom surface, two handles arranged perpendicular to said bottom surface and opposite each other, a locking means for holding said cell culture flask, and a heat transfer pad arranged on the bottom surface of said holder.

12. 12. The set of claim 11, wherein the cell culture flask holder further comprises at least two magnets disposed on an exterior of a bottom surface of the cell culture flask holder that provide a mounting means for the culture shaker that includes a permanent electromagnet.

13. 12. The set of claim 11, wherein the locking means of the cell culture flask holder comprises a clamping means and a safety latch disposed on a bottom surface of the cell culture flask holder.

14. 12. The set of claim 11, wherein the locking means of the cell culture flask holder comprises four guillotines each holding a toothed jaw and positioned on a bottom surface of the cell culture flask holder.

15. Use of the set according to any one of claims 1 to 14 for maintaining and transporting at least one cell culture flask arranged in at least one cell culture flask holder compatible with said set.

16. A culture shaker comprising the set according to claim 1.

17. a liquid handling system, at least one robotic arm, at least one automated hotel, an automated refrigerated incubator, an automated microplate centrifuge with an automated loader, a cell counter and at least one shaking incubator, and optionally an automated orbital shaking incubator; 2. An automated cell passaging platform, characterized in that at least one said orbital shaking incubator is designed to be arranged with said set for maintaining and transporting at least one cell culture flask according to claim 1.

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