How to defrost frozen bags

Abstract

This document describes a consistent and repeatable thawing method for frozen samples within a bag storage container. The method and system can allow the use of multiple bag storage container sizes within the same device. A subset of multiple sensors can be adapted to the thawing method. The method may further include heating the frozen sample using a first heater row and a second heater row. Furthermore, the method may include measuring multiple second temperatures of the bag container. After a second threshold temperature or slightly exceeding a second threshold temperature, the method may include heating with the first heater row and then terminating heating with the second heater row. The method may include terminating heating of the partially thawed sample with the first heater row after a period of time has been achieved by heating the partially thawed sample with the first heater row.

Description

[0001] Cross-reference to related applications

[0002] This application claims priority to U.S. Provisional Application No. 62 / 962,733, filed January 17, 2020, the entire contents of which are incorporated herein by reference for all purposes. Background Technology

[0003] Cryopreservation of cells in suspension is a recognized and accepted technique for the long-term archiving storage and resuscitation of live cells. As a general method, cells are suspended in a cryopreservation medium, which typically includes a saline solution, buffers, nutrients, growth factors, proteins, and cryopreservatives. The cells are then dispensed into archiving storage containers of the desired size and volume, and the containers are then cooled until their contents are frozen. Typical long-term archiving conditions include liquid nitrogen vapor storage, where temperatures are typically between -196°C and -150°C. Vial or pouch storage containers can be used to store such sample fluids, as well as cryopreserved samples suitable for a given volume or application.

[0004] Successful revival of live cells preserved in this way depends on minimizing the growth of harmful ice crystals in the intracellular region during freezing and thawing. Reviving a sample from a cryogenic archiving state involves thawing the sample to a completely liquid state. During thawing, the rate of temperature change can affect the viability of cryogenically preserved cells. The solid contents of the sample storage container contain numerous islands of crystallized water, interspersed with channels of glassy aqueous solutes mixed with small ice nuclei. During the transition from cryogenic storage temperature to the fully liquid state, there is an opportunity for water molecule rearrangement within the sample, including thermodynamically favorable extension of small ice nuclei within the cells. Since the growth of intracellular ice crystals carries the potential for cell damage, and since the extent of crystal growth varies over time, minimizing the transition time during the phase transition is desirable.

[0005] Rapid temperature transitions in the sample container are typically achieved by partially immersing the container in a water bath set at approximately 37°C. While faster thawing rates can be achieved by increasing the bath temperature, immersing the container in the bath establishes a temperature gradient within the container, with the highest temperature located at the container walls. As a result, a transient thermodynamic state occurs, where the temperature of the liquid-solid mixture exceeds its melting temperature even with frozen material present in immediate proximity. Therefore, the temperature gradient within the container sets an upper limit on the bath temperature. Furthermore, due to the known toxic effects of common cryoprotectants on cells, the temporal and temperature differences in cell exposure in the liquid state allow for changes in cell viability as the thawing process completes. Since the toxic effects of cryoprotectants are enhanced at higher temperatures, lower liquid temperatures are desirable. For this reason, some thawing protocols typically include a rapid thawing phase that terminates while a small amount of solid material remains in the sample container. Upon removal from the water bath, the sample temperature rapidly equilibrates to near the phase transition temperature. Thawing protocols typically seek to minimize the duration for which the thawed sample remains in a concentrated state of cryoprotectant, and usually apply subsequent steps such as diluting the sample or replacing the cryopreservation medium with a culture medium within the shortest possible time interval.

[0006] While some thawing methods have been proposed to automate sample thawing, further improvements can be made, particularly regarding samples stored in bag-type cryopreservation containers (referred to as "freezing bags"). Summary of the Invention

[0007] This disclosure generally relates to systems and methods for thawing frozen samples within bagged storage containers. In various aspects, this disclosure relates to the cryopreservation of cells, tissues, and fluids, and to systems, apparatus, and methods for resuscitating cryopreserved cells, tissues, and fluids.

[0008] Embodiments of the present invention allow for consistent and repeatable thawing of frozen samples within bag storage containers. Thawing can achieve just enough to retain some solid phase in the liquid phase. Embodiments may allow the use of multiple bag storage container sizes in the same apparatus.

[0009] Embodiments of the present invention may include a method for thawing a frozen sample within a bag container. The method may include measuring a plurality of first temperatures of the bag container in contact with a first surface and a second surface. Each of the plurality of first temperatures may be measured by a different sensor among a plurality of sensors. Each of the plurality of sensors may be configured to measure temperature at a different location within the bag container. The method may include comparing each of the plurality of first temperatures to a first threshold. The method may further include using the comparison to determine a subset of the plurality of sensors, wherein each sensor in the subset of the plurality of sensors measures a first temperature below the first threshold. The method may further include heating the frozen sample using a first heater array and simultaneously heating the frozen sample using a second heater array. Furthermore, the method may include using a subset of the plurality of sensors to measure a plurality of second temperatures of the bag container. A second temperature exceeding a second threshold may indicate that a partially thawed sample is present in the bag container. At or slightly after this level, the method may include heating the partially thawed sample using the first heater array and terminating heating of the partially thawed sample using the second heater array. The method may include terminating the heating of the partially thawed sample using the first heater row after the sample has been heated using the first heater row for a certain period of time.

[0010] The embodiment may also include a defrosting system. The defrosting system may include defrosting devices. The defrosting devices may include a first surface, a second surface, multiple sensors, a first heater array, and a second heater array. The defrosting system may also include a computer system. The computer system may include instructions that, when executed, control the defrosting devices to perform the defrosting method.

[0011] A better understanding of the nature and advantages of the embodiments of the present invention can be obtained by referring to the following detailed description and accompanying drawings. Attached Figure Description

[0012] Figure 1A and Figure 1B A view of a sample thawing apparatus focused on a plate, heater, and sensor according to an embodiment of the present invention is shown.

[0013] Figure 1C An exploded view of a sample thawing apparatus focusing on a plate, heater, and sensor according to an embodiment of the present invention is shown.

[0014] Figure 2 A method for thawing frozen samples in a bagged container according to an embodiment of the present invention is shown.

[0015] Figure 3 A view of a sample thawing apparatus according to an embodiment of the present invention is shown, wherein the top of the outer shell has been removed.

[0016] Figure 4 The image shows a top right front view of the device according to an embodiment of the invention, wherein additional parts of the device have been selectively removed to concentrate the temperature sensor in the heater plate.

[0017] Figure 5 A cross-sectional view of a thermal sensor according to an embodiment of the present invention is shown.

[0018] Figure 6 A computer system according to an embodiment of the present invention is shown. Detailed Implementation

[0019] To thaw cells stored in cryopreservation bags, standard practice involves rapidly heating the cells in a warm water bath (e.g., 37°C) to just about the last bit of ice about to melt, and then slowly diluting the cells into growth medium. If the sample is allowed to become too hot, the cells can begin to metabolize, and the sample can become poisoned by dimethyl sulfoxide (DMSO), which is often used in the freezing process. Typically, the thawing of cryopreserved cells and tissues is performed by laboratory technicians, and the protocols applied can vary not only between individual technicians but also depend on the technique. The completion of sample thawing is often subjectively judged by each individual technician and can lead to variations in the thawing rate or the amount of sample allowed to become too hot. While reproducible thawing profiles can theoretically be achieved using a bath and manually controlled cryopreservation, the anticipated variations in both technique and protocol compliance, especially combined with the requirement to frequently remove cryopreservation bags from the bath to monitor thawing status, make deviations from the standard curve nearly deterministic. Removing the freezer bag from the bath disrupts the heat transfer from the bath water to the freezer bag, and visual assessment of the thawed state is often difficult and can be complicated by the presence of labels and printed writing surfaces that are integrated features of the freezer bag product. Furthermore, the water bath is a source of contamination, and unintentional immersion of the freezer bag seal can introduce bath fluid into the freezer bag contents during opening or removing the seal.

[0020] Embodiments of the present invention allow for consistent and repeatable thawing of frozen samples in freezer bags. Multiple freezer bag sizes can be thawed in the same device. Some freezer bags may have a size substantially smaller than the heating plate used for thawing. Embodiments of the present invention avoid over-thawing and sample damage by tilting downwards or shutting off heaters positioned away from freezer bags that are significantly smaller than the heating plate. The latent heat in the heating plate is sufficient to thaw a portion of the freezer bag. Other heaters can continue thawing another portion of the freezer bag positioned in contact with or more closely with the heaters.

[0021] I. Overview

[0022] The thawing system can be designed to thaw the frozen contents of standard freezer bags (which are a type of bag container). Because freezer bags are commercially available in nominal volumes ranging from 25 ml to 1000 ml and larger, but in most cases used to contain volumes that are a portion of the nominal size, the maximum cross-sectional thickness of the bag when frozen in a flat orientation is approximately 0.4 inches. Freezer bags are typically rectangular in outline and are provided with a tubing system through which the bag is filled. After filling and before freezing, the filling tubing is heat-sealed and trimmed so that the tubing protrudes no more than approximately one inch from the end of the bag. In addition to the tubing, the same end region of the freezer bag will typically have two or more port features, which are sealed until broken by the connector tubing fittings, through which the contents of the bag can be retrieved. The area where the ports of the freezer bag are located is referred to as the neck or proximal end of the freezer bag. The thawing system can accept frozen bags and contents with initial temperatures ranging from approximately -70°C to -196°C, and rapidly increases the temperature of the bags and contents through phase change temperature until the contents are primarily in a liquid state. Since the thawing contents, as containers, do not have visual indications of unacceptably high temperatures, the cryopreservation industry has adopted the expectation of some solid residue at the end of the thawing process as a matter of course.

[0023] The thawing system can accept a range of frozen bag sizes, with a range of filling volumes applicable to each bag size. The instrument can thaw the bag contents at a rate close to that experienced when the bag is immersed in a water bath at approximately 37°C. The thawing system can agitate the bag contents during the thawing process to maintain a uniform temperature distribution and can automatically terminate the thawing process when some solid phase remains in the bag.

[0024] Figure 1A , Figure 1B and Figure 1CThe defrosting unit 100 is shown with plates, a heater, and a sensor. The defrosting unit 100 may include two aluminum plates (e.g., a lower plate 104 and an upper plate 108) that contact the upper and lower main surfaces of a frozen bag with a light clamping pressure to ensure optimal contact between the plates and the bag. The lower plate 104 may be configured as an extendable drawer and thus includes an area to accommodate the largest bag size (e.g., 1000 ml). The frozen bag may be positioned between the lower plate 104 and the upper plate 108, with the port of the frozen bag located on the left side of the plates relative to these figures. The upper plate 108 may be hinged on an axis parallel to the long dimension of the frozen bag, allowing the plate to rock to mix the contents of the bag during the defrosting process. Furthermore, the upper plate 108 may be confined to areas of the frozen bag where no port features are present, such that the ports do not restrict the clamping pressure of the upper plate on the portion where the contents of the frozen bag are located. Thus, ports for all bag sizes can be intentionally positioned in the same area of ​​the lower plate, such that as bag size and capacity increase, the bag extends into areas of the plate away from the port area. For convenience, "top," "bottom," "lower," and "upper" are used to describe the orientation of bags or tiers. However, freezer bags and tiers can be oriented vertically or at any angle between horizontal and vertical.

[0025] The lower plate 104 may include a group of thermal sensors 112 (sensors 112a-112e) embedded in an insulating medium, such that when a freezer bag is placed on the lower plate 104, the sensors 112a-112e can be thermally coupled to the bag but not to the lower plate 104. The sensors 112a-112e can report the temperature of the bag surface, which deviates only a few degrees from the contents of the freezer bag on the inner wall opposite the sensors. Because the plate sensors report near-proximity temperatures of the bag contents and because the mixing system promotes uniform temperature within the freezer bag contents, termination of the thawing process can be triggered when a preset temperature threshold is reached. This preset temperature threshold has been pre-determined to be associated with a phase transition state in which a low proportion of solid phase remains in the bag solution. The thermal sensor group 112 may be distributed along a line in the lower plate 104. The thermal sensor group 112 may be distributed such that a freezer bag of a given size typically contacts two or more sensors. Due to different freezer bag geometries, the sensors 112a-112e may be unevenly distributed across the plate. The thawing termination temperature threshold can be configured such that all participating temperature sensors must pass through this threshold to terminate thawing. As a safety mechanism, temperature readings from sensors that have exceeded the threshold can be used to monitor secondary temperature thresholds for over-temperature and to trigger termination of the thawing process to prevent overheating into potentially harmful temperature ranges to the viability of the biological suspension within the frozen bag solution.

[0026] The set of defrosting control variables may include a temperature value that defines a threshold below which a given bag temperature sensor must reach to be suitable for participation in the defrosting control process. As an example, the lower plate 104 may include five separate temperature sensors (e.g., sensors 112a, 112b, 112c, 112d, and 112e) strategically distributed along the centerline of the lower plate 104, such that they also contact the frozen bag along the centerline. Since the length of the bag varies depending on its nominal capacity, some of sensors 112a-112e will not engage with the bag for lower capacity bags. Sensors capable of engaging with a frozen bag of a given capacity can be identified in the variable table or dynamically by the absence of a temperature drop at the start of the defrosting cycle, by means of their location. However, if the lower surface of the frozen bag is not flat or includes accidental cavitation or any depressions, then sensors that would otherwise report the frozen bag temperature may provide false information. For this reason, each sensor may need to report a temperature drop with a sufficient value to conform to the defrosting process control algorithm. If no sensor meets the temperature drop threshold after a specific time limit, this fault status can be notified to the user via the display screen. The time limit value for this number of seconds after the start of the thawing process can be stored in a set of control variables. The failure event can be incorporated into an algorithmic sequence that terminates the thawing process and removes the frozen bag from the instrument.

[0027] The data stream from the temperature sensor in the freezer bag can be stored and monitored by a graphical display, or transferred along with metadata for a specific thawing file to a portable medium for archival storage and inspection.

[0028] A. Thawing stage

[0029] The thawing process of a frozen bag can be divided into two stages. The first stage can be the temperature transition stage, where the heat flow into the bag and its contents primarily serve to transition the temperature from the initial low temperature to the onset of a phase change in the frozen bag contents. The onset of the phase change can be characterized by the initial formation of a liquid phase within the frozen bag. Since the contents of a frozen bag typically include complex aqueous formulations, the phase change is not a sudden transition but rather a diffusion over a temperature range. When the phase change begins during the thawing process, the liquid phase can distribute throughout the internal surface of the frozen bag, thereby increasing the thermal contact between the bag and the heating plate as the bag conforms to the plate surface. The interval from the onset of the phase change to the end of the thawing process can be defined as the second stage.

[0030] The upper plate 108 and lower plate 104 may each include two rows of flat resistance pad heaters. On the lower plate 104, a first row 116 of heaters may be located at the proximal or neck region of the bag, while a second row 120 of heaters with a higher wattage may be positioned distal to the first row 116 relative to the neck and port regions of the frozen bag. On the upper plate 108, a third row 124 of heaters may be located at the proximal or neck region of the bag, while a fourth row 128 of heaters with a higher wattage may be located distal to the third row 124. Although the number of heaters may be fewer or more in embodiments, each row is shown as including two identical heaters. Using only one heater row per plate can lead to over-thawing of the distal portion of the bag. Using too many heater rows per plate can lead to unnecessary process complexity and increased cost. However, in some embodiments, additional heater rows (e.g., a total of 3, 4, 5, 6, 7, 8 or more) may be used. Further division of the distal heater rows can be used for segmented bags or bags with diaphragm boundaries.

[0031] Because the pad heater can have a uniform energy output per unit area, a larger wattage heater covers a wider area compared to a smaller wattage neck heater. For a range of freezer bag sizes, as the size of the freezer bag decreases, the larger areas of both the upper and lower plates will lack heat dissipation zones, and thus will begin to increase in temperature at a rate greater than the area of ​​the plate in contact with the freezer bag. The end result of this temperature imbalance in the plates can be an increased heat flux into the distal end of the freezer bag, and therefore a phase transition that can occur at a greater rate than in the neck region. If the defrosting algorithm terminates defrosting based on high sensor readings from the distal region, the neck region may still be in the solid phase, and therefore the defrosting process at termination will be non-uniform.

[0032] B. Phase Transition Control

[0033] To establish a balanced thawing endpoint for different freezer bag sizes, the system must determine the freezer bag size. The freezer bag size can be determined dynamically via sensors or from user input. Size determination can identify table entries for a set of variables that will control the thawing process for a specific bag size. The variable table may include time point values ​​that can be used to trigger the transition from the first thawing stage to the second thawing stage. Stage transitions can also be controlled by a temperature setpoint, which can be triggered by any combination of data streams from qualified freezer bag temperature sensors. The neck heater rows and distal heater rows on the upper and lower plates can be individually controlled by local temperature sensor feedback loops using a PID algorithm to achieve and maintain the setpoint temperature. An initial or first-stage temperature setting can be applied to all heater rows to achieve a rapid freezer bag temperature transition to the start of the phase change.

[0034] After the phase change begins, a second set of temperature settings can be applied to the neck heater rows (e.g., first row 116 and third row 124) and the distal heater rows (e.g., second row 120 and fourth row 128) to balance the phase change process of the frozen bag contents along the length of the frozen bag. Heater setting changes for medium and small-sized bags can be stored in a variable table. The variable table can be based on empirical data and can be used to input control variables. The control variables can be adjusted to optimize the thawing process outcome. By way of example, a 250ml nominal-size frozen bag can hold biological material contents ranging from 30ml to 70ml, and the frozen bag can occupy approximately half of the plate area. During the first stage of the thawing process, the proximal or neck portion of the plate, as well as the distal portion, is temperature-controlled to the first stage set point. However, under heat sink loading, both plates may experience a temperature drop. Due to the temperature drop, the heater rows can be activated in response to the temperature drop in an attempt to restore the plates to the temperature set point.

[0035] During the second stage of defrosting, as the temperature of the frozen bag approaches the predetermined temperature endpoint, the demand on the heating system can be reduced to restore the plate temperature to the setpoint. However, the distal portion of the plate, which is not in contact with the frozen bag, may contain a significant amount of residual heat, which can migrate towards the proximal end of the plate, towards the heat dissipation area of ​​the frozen bag. Without compensation, an additional distal area plate heat sink could cause the distal area of ​​the frozen bag to defrost before the proximal area. Therefore, a second-stage heater setting can be performed at this point to switch the distal heaters to a lower temperature setting, resulting in a more uniform heat distribution across the plate. In some cases, the proximal heater array can be switched to a higher temperature setting, allowing the PID control loop to apply a larger energy inflow. For a given bag solution, the temperature setpoints for all bag sizes can be optimized through prior testing. In addition to the stage plate temperature setting, the idle plate temperature setting can be selected as a hold temperature before and between defrosting procedures. The transition from the first to the second stage plate temperature setting can be triggered by temperature data provided by the frozen bag temperature sensor, by the time the setting is initiated after defrosting, or by a combination of both controls.

[0036] The defrosting stage and idle plate temperature settings, stage transition signal settings, sensor threshold identification, defrosting completion temperature, and high-temperature limit can be stored as profiles for specific defrosting applications. Profiles may also include other control values ​​for instrument operation, such as target plate clamping pressure values ​​and absolute defrosting duration values, to provide safety checkpoints to prevent unintended fault conditions from causing over-defrosting or over-temperature states. Profiles can be stored in a database for easy retrieval and loading into defrosting control variables to execute desired defrosting conditions. Defrosting profiles can be transferred to portable storage media or cloud storage, and external defrosting profiles can be transferred from portable storage media or cloud storage to the instrument profile storage database.

[0037] II. Example Method

[0038] Figure 2 An example method 2000 for thawing frozen samples in a bag container is shown. The bag container can be any freezing bag described herein. The freezing bag can have a nominal size of 25, 50, 250, 500, 750, or 1,000 ml. Method 2000 may include the use of any system described herein. Method 2000 may further include details of the thawing process described above.

[0039] The bag container may include a port. The port of the bag container may be positioned immediately adjacent to a first end of a first surface and a first end of a second surface. The bag container may have a surface area smaller than the surface area of ​​the first surface and may have a surface area smaller than the surface area of ​​the second surface. The surface area of ​​the first surface may be equal to the surface area of ​​the second surface. The surface area of ​​the bag container may be from 10% to 20%, 20% to 30%, 30% to 40%, 40% to 50%, 50% to 60%, 60% to 70%, 70% to 80%, 80% to 90%, or 90% to 95% of the surface area of ​​the first or second surface. The length of the bag container (excluding the port) may vary from 1 to 2 inches, 2 to 3 inches, 3 to 6 inches, 6 to 8 inches, 8 to 12 inches, or greater than 12 inches. These surfaces may be longer than the longest bag container used for defrosting, yet still accommodate much smaller bag containers without over-defrosting their contents.

[0040] The first and second surfaces may be horizontally oriented such that one surface is the bottom surface and the other is the top surface. The first and second surfaces may also be vertically oriented or oriented at an angle between horizontal and vertical. One plate may be larger than the other. In some embodiments, the surface area of ​​the first surface may be 0% to 10%, 10% to 20%, 20% to 30%, 30% to 40%, 40% to 50%, or 50% to 100% larger than the surface area of ​​the second surface. The surface may be any surface or plate described herein. Any thermally conductive surface may be used, including flexible materials, and may be or may comprise gels, liquids, fabrics, nonwovens, or meshes.

[0041] The bag-type container may overlap with the first surface at a first portion and may not overlap with the first surface at a second portion. Similarly, the bag-type container may overlap with the second surface at a first portion and may not overlap with the second surface at a second portion. A first heater row may be positioned closer to the first portion of the first surface and the second portion of the second surface than the second portion of the first surface and the second portion of the second surface. A second heater row may be positioned closer to the second portion of the first surface and the second portion of the second surface than the first portion of the first surface and the second portion of the second surface.

[0042] In one embodiment, the first heater row may be positioned closer to the first end of the first surface and the first end of the second surface than the second heater row is positioned at the first end of the first surface and the first end of the second plate. In another embodiment, both the first and second heater rows may be located on the first surface and configured to provide temperature readings for a contacted bag-like container rather than the first surface. In these and other embodiments, the second surface may include a third and a fourth heater row. The third heater row may be aligned with the first heater row, and the fourth heater row may be aligned with the second heater row. The first heater row may be identical to the third heater row. The second heater row may be identical to the fourth heater row. Each heater row may include one, two, three, four, or more heaters.

[0043] In one embodiment, a line perpendicular to the main surface of the first surface (e.g., a plate) extends through the second heater row but not through the bag container. The first surface may be parallel to the second surface. In some embodiments, a line perpendicular to the main surface of the first surface (e.g., a plate) extends through the second heater row and through the bag container.

[0044] Method 2000 may include receiving user input specifying the size of a bag container. In some embodiments, the only input received from the user is the size of the bag container. Method 2000 may include receiving user input specifying the type of medium in the bag container. In some embodiments, method 2000 may include loading the bag container into a drawer of a device. Method 2000 may include contacting the bag container with a first surface of a sample thawing device. Furthermore, method 2000 may include contacting the bag container with a second surface. Method 2000 may further include clamping the bag container between the first and second surfaces.

[0045] At block 2002, method 2000 may include measuring a plurality of first temperatures of a bag container in contact with a first surface and a second surface. Each of the plurality of first temperatures may be measured by a different sensor among a plurality of sensors. Each of the plurality of sensors may be configured to measure the temperature at a different location on the bag container. The plurality of sensors may include 5 sensors, 2 to 5 sensors, 5 to 8 sensors, 8 to 10 sensors, or more than 10 sensors. The plurality of sensors may be unevenly distributed on the first surface and the second surface. The bag container may be in contact with at least 2, 3, 4, or more sensors. One surface may include sensors, while the other surface may not have any sensors. In some embodiments, sensors may be present on both surfaces.

[0046] At block 2004, method 2000 may include comparing each of a plurality of first temperatures to a first threshold. The first threshold may be lower than the temperature of the bag container measured when the bag container initially contacts the first and second surfaces. The first threshold may depend on the size of the bag container and / or the medium. For example, the first threshold may be -10°C, from -30°C to -20°C, from -20°C to -10°C, or from -10°C to 0°C. In some embodiments, the first threshold may be a temperature difference, and the measured first temperature may also be a temperature difference. As an example, the first threshold may be a temperature change of -15°C. The first threshold may be specific to a certain duration. For example, the duration may be 30 to 45 seconds, 45 to 60 seconds, or 1 to 2 minutes after or before the freezer bag is clamped onto the two surfaces.

[0047] At block 2006, method 2000 may include using comparison to determine a subset of multiple sensors. The subset of multiple sensors may be considered suitable for the thawing process. Each sensor in the subset of multiple sensors may have measured a first temperature below a first threshold. The subset of multiple sensors may include fewer sensors than a plurality of sensors. In some embodiments, the subset of multiple sensors may be a plurality of sensors.

[0048] The subset can be determined using the size of the bag container. The size of the bag container can be received as user input or determined by sensors. Sensors determined to be outside the area of ​​the bag container can be excluded from the subset of multiple sensors.

[0049] At block 2008, method 2000 may include heating a frozen sample using a first heater row and simultaneously heating the frozen sample using a second heater row. Heating the frozen sample may occur before, simultaneously with, or after measuring multiple first temperatures. Heating the frozen sample using the first heater row may include setting a surface temperature setpoint for the first and second heater rows within the range of 37°C to 45°C, 37°C to 40°C, 40°C to 42°C, 42°C to 45°C, 45°C to 50°C, or exceeding 50°C. This temperature may be the surface temperature measured separately from multiple sensors measuring the temperature of the bag container. The first and second heater rows may have the same temperature setpoint. Heating may be controlled by a PID loop or any suitable control loop. Heating may also include a third and fourth heater row. Method 2000 may include stirring the surfaces and the bag container during heating.

[0050] At box 2010, method 2000 may include using a subset of multiple sensors to measure multiple second temperatures of the bag container. The multiple second temperatures may be associated with thawing time or duration. A temperature distribution can be determined from the measured temperatures and time.

[0051] At block 2012, method 2000 may include heating a partially thawed sample using a first heater array, while terminating heating of the partially thawed sample using a second heater array when a second temperature among a plurality of second temperatures exceeds a second threshold. The second threshold may be in the range of 0°C to 8°C, including 0°C to 4°C. In some embodiments, the second threshold is a temperature difference, and the measured second temperature may be a temperature difference. The temperature difference may be the difference from a first threshold. In some embodiments, only one of the plurality of second temperatures needs to exceed the threshold. This one second temperature may be, as appropriate, the first second temperature or, as appropriate, the last second temperature to exceed the threshold. In some embodiments, two second temperatures, a majority of the second temperatures, or all of the second temperatures need to exceed the threshold. In some embodiments, the second threshold may be a temperature array. For example, the second threshold may include a value for the highest second temperature and a value for the lowest second temperature. In some embodiments, the average or median of the plurality of second temperatures needs to exceed the threshold. Furthermore, the second threshold may be a temperature array. For example, the second threshold may include a value for the highest second temperature and a value for the lowest second temperature. If any, all, or some values ​​in the array exceed the threshold, it is considered that the second threshold has been exceeded. The second threshold can be determined empirically and can be determined using the size of the bag container and / or the medium.

[0052] Terminating heating of a partially thawed sample using the second heater row can include lowering the temperature setpoint of the second heater row. Lowering the temperature setpoint below the heater row temperature effectively terminates heating. However, if the surface temperature drops below the temperature setpoint, the second heater row can be reactivated.

[0053] At block 2014, method 2000 may include terminating heating of the partially thawed sample using the first heater row after a period of time has been achieved using the first heater row. In some embodiments, when heating of the partially thawed sample using the first heater row is terminated, the majority of the partially thawed sample is an aqueous solution and the solid phase remains in the partially thawed sample. For example, 70% to 80%, 80% to 90%, 90% to 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%, or 99.99% of the contents, by mass or volume, may be a liquid phase, with the remainder being a solid phase. The remaining solid phase may be particulate. The solid phase may be fine particles that are substantially uniformly distributed through a bag-like container. The partially thawed sample may resemble a fine slurry.

[0054] In some embodiments, termination of heating of the partially thawed sample may occur at a predetermined time. The predetermined time can be calculated using empirically derived phase transition duration. The predetermined time may be 30 seconds to 1 minute, 1 to 3 minutes, 3 to 5 minutes, 5 to 7 minutes, 7 to 10 minutes, 10 to 15 minutes, or more than 15 minutes after reaching a first threshold time. In some embodiments, the predetermined time may be a safety check to prevent overheating in the event of a malfunction or anomaly.

[0055] In some embodiments, method 2000 may further include measuring multiple third temperatures of the bag container using a subset of multiple sensors. When one of the multiple third temperatures exceeds a third threshold, heating of the partially thawed sample using the first heater array is terminated. The third threshold may be in the range of 4°C to 25°C. The third threshold may be determined empirically and may vary based on the bag container size and / or medium. Similar to the second threshold, the third threshold may be a temperature difference. The third temperature may also be a temperature difference. Furthermore, similar to the second threshold, the third threshold may be based not only on third temperatures but may include multiple third temperatures or statistical values ​​of multiple third temperatures. Additionally, similar to the second threshold, the third threshold may be a temperature array. For example, the third threshold may include a highest third temperature value and a lowest third temperature value. If any, all, or some values ​​in the array are exceeded, the third threshold may be considered exceeded. For example, if the minimum third temperature is higher than 4°C or the maximum third temperature is higher than 12°C, the third threshold may be considered exceeded.

[0056] In some embodiments, multiple third temperatures can be compared to a reference temperature profile. When one of the multiple third temperatures deviates significantly from the reference temperature profile, a first heater is used to heat the partially thawed sample. The deviation from the reference temperature profile can also indicate leakage, overfilling, incomplete filling, or abnormality of the bag container or device. Heating can be based not only on the third temperatures but can also include multiple third temperatures or statistical values ​​of multiple third temperatures, similar to what is described for other thresholds.

[0057] Method 2000 may include removing the bag container from the device after terminating heating of the partially thawed sample using the first heater array. In some embodiments, a drawer including the first surface may automatically open after heating is terminated. The solid phase in the bag container may become a liquid phase within 10 to 20 seconds, 20 to 30 seconds, or 30 to 60 seconds after removal from the first surface.

[0058] In some embodiments, method 2000 may include thawing a second frozen sample in a second bag-shaped container in contact with the first and second surfaces. The second bag-shaped container may have a different size than the first bag-shaped container. The thawing process of the second bag-shaped container may include any of the methods described for thawing bag-shaped containers.

[0059] III. Example System

[0060] Example systems that can be used with the methods described in this article may include BioLife Solutions. The entire contents of the CB (Bio-Lay Cell Procedural Resuscitation System) and the system described in U.S. Application No. 16 / 054,454, filed August 3, 2018, are incorporated herein by reference for all purposes. Example systems may be used with... Figure 1A , 1B It can be combined with and may include aspects of the means set forth in 1C.

[0061] Figure 3The internal components of the defrosting unit 100 are shown, specifically a portion of the defrosting unit 200, including the upper housing, touchscreen cover, touchscreen interface, and the removed front panel. The rear panel 135 is shown. An upper heater plate 240 is shown, which may be formed of a material with generally high thermal conductivity, such as, but not limited to, aluminum, aluminum alloys, copper, copper alloys, stainless steel, carbon fiber, graphene, etc. In such respects, the upper heater plate 240 may have a thermal conductivity from 12 to 400 watts / (m·K) (κ = 12-400 W / (m·K)), or increments or gradients of thermal conductivity within that range. Two or more heaters 255 are joined to the upper surface of the upper heater plate 240. In some aspects, the heaters 255 may be resistance heaters, such as flexible silicone pad heaters. In other aspects, the heaters 255 may cover a large portion of the surface area of ​​the upper surface of the upper heater plate 240. In some embodiments, two heaters 255 are separated by a gap 256, which allows one or more thermistors 257 to pass through or be placed therebetween. One or more thermistors 257 may be embedded in the upper heater plate 240 to monitor the temperature of the upper heater plate 240 and to provide thermal measurement information to a control unit, enabling temperature control via feedback circuitry of the control unit, which adjusts the energy applied to the two heaters 255. In some embodiments, the heaters 255 on or within the upper heater plate 240 may operate on independent control circuitry, allowing for proactive adjustment (manually by operator input or automatically by algorithm) of the heat input to the heater plate in response to non-uniform heat sinks that may contact the upper heater plate 240, to balance the temperature of localized areas of the upper heater plate 240. In some embodiments, the number of individual heaters 255 on the upper plate may be greater than two to exert greater regional control over the temperature of the upper heater plate 240, thereby providing more precise control and dynamic balancing of the heat input to the plate, allowing the defrosting unit to accept bagged containers and defrost uniformly over a wide range of sizes or volumes.

[0062] In some aspects, heaters 255 can be controlled individually as a subset based on their position on the upper surface of the upper heater plate 240. For example, heaters 255 positioned towards the front of the upper heater plate 240 can be adjusted to a different temperature than heaters positioned towards the rear of the upper heater plate 240. Alternatively or in combination, heaters 255 positioned towards the left side of the upper heater plate 240 can be adjusted to a different temperature than heaters positioned towards the right side of the upper heater plate 240. In other aspects, heaters 255 can be insulated from each other on the surface of the upper heater plate 240 and / or from the rest of the defrosting device.

[0063] The upper heater plate 240 is attached to the cantilever assembly 202 via two concentric ring surface bearings, which interface with three flanged dry bearings 250 that rotate on an axis embedded in each of the two cantilevers 205. The cantilever assembly 202 can be considered as a cantilever clamping mechanism formed by the two cantilevers 205, a cross plate 210, and a push rod 215. Circular bearing races 245 restrict the movement of the upper heater plate 240 to rotation about an axis passing through the intersection of a horizontal plane passing through the middle of the freezer bag and a front vertical plane passing through the centerline of the freezer bag. The two cantilevers 205 are connected by two cross plates 210, which stiffen the cantilever assembly and prevent twisting under uneven clamping loads. The cantilevers 205 rotate on two pivots 225, which are connected to two bearing seats (see below) also attached to the base plate 285. Figure 4 The cantilever assembly 202 is hinged as a whole by push rod 215, which is attached to the two cantilever 205 via two pivot bearings 220. Push rod 215 is hinged by the force generated by the screw jack mechanism 290.

[0064] A model of a standard cryogenic storage bag is shown as a container 260 sandwiched between an upper heating plate 240 and a lower heating plate 230. The lower heating plate 230 rests in a frame receiving seat 235, which is part of and movable with an extendable drawer 265. The lower heating plate 230 may be formed of the same or different materials having the corresponding thermal conductivity as described with respect to the upper heating plate 240. When the clamping pressure generated by the screw jack mechanism 290 is released and the cantilever assembly 202 and the attached upper heating plate 240 are raised, the drawer 265 (and the container 260 placed in the drawer) extends forward freely from the remainder of the instrument on the rolling bearing track. The forward and retraction of the drawer 265 allows any suitable storage bag (here represented by container 260) to be introduced into and removed from the instrument.

[0065] Further shown is a graphics control circuit board 280, attached to the upper housing, which includes various non-transient computer-readable media. The graphics control circuit board 280 can be electronically connected to the touchscreen interface 140 and / or access port 145. An external circuit board 295, also including various non-transient computer-readable media, includes a microcontroller that manages the instrument's mechanical and electrical power components. The defrosting unit 200 may further include a power cord and power switch interface module 270, a cooling fan 292, and a wiring link harness 267 (shown as volume-filled raw components).

[0066] Figure 4The internal components of the defrosting unit 200 are shown, focusing on temperature sensors integrated into the structure of the lower heater plate 230. Specifically, multiple container temperature sensor islands 675 are present and extend through the body of the lower heater plate 230, allowing the temperature sensor islands 675 to monitor the surface temperature of cryogenic storage containers resting on or sandwiched against the surface of the lower heater plate 230. The multiple temperature sensor islands 675 are distributed along the lateral centerline of the lower heater plate 230. The spacing between the temperature sensor islands 675 is typically such that two or more temperature sensor islands 675 contact a standard cryogenic storage bag container, representing most commercially available bag container products. In some aspects, such as in the case of a 25 ml storage bag with a relatively small surface area, only one of the temperature sensor islands 675 may contact the storage bag. In some embodiments, two temperature sensor islands 675 may be distributed and sized to even contact the 25 ml storage bag. In contrast to the upper heater plate 240, the location of the temperature sensor island 675 within the lower heater plate 230 offers the advantage that, due to gravity, the container contents will displace any air pockets present in the container interior on the bottom side of the container upon reaching the liquid phase, thereby providing an optimal thermal path between the contents and the temperature sensor island 675. In other words, any air bubbles in the freezing bag will rise to the top of the bag, and the bottom of the bag will be substantially flat on the surface of the lower heater plate 230, maximizing contact with the available thermal sensor. Another advantage of mounting the sensor in the lower heater plate 230 is that the lower heater plate 230 remains stationary, while the sensor mounted in the upper heater plate 240 will experience oscillations during a portion of the thawing process. Of course, in some embodiments, it should be understood that a thermal sensor positioned in the upper heater plate 240 may provide alternative advantages, such as targeting a specific location for thermal measurement on a particular container. In other embodiments, the temperature sensor may be a thermocouple, a thermistor, an IR sensor, or an RTD sensor.

[0067] Another aspect of the entire sample thawing apparatus may include a communication module formed of a non-transient computer-readable medium and configured to transmit sample data, including thermal data regarding the sample container held by the thawing apparatus, to other devices. The communication module may be directly coupled to a temperature sensor of the sample thawing apparatus (such as temperature sensor island 675). The communication module may also be electrically coupled to a graphical control board 280, an external board 295, and a touchscreen interface 140, thereby allowing control of all aspects of the instrument. The communication module may be further configured to communicate with a remote microprocessor (e.g., a cloud-based server or computer) to classify and display data. The communication module is also configured to receive instruction data or vial identification data and control the heating of the vials accordingly.

[0068] Figure 5 A schematic cross-sectional view of the lower heater plate sensor 800 (which may alternatively be referred to as a "thermal sensor") is shown, illustrating a sub-component of the sensor island. An insulating disk 810 rests in a cylindrical recess within the lower heater plate 230, supported by an annular flange 805, which may be an integral part of the lower heater plate 230. In some aspects, the insulating disk may be adhered to a thermally conductive material, which in turn is attached to a thermocouple or other temperature sensor structure. In some embodiments, the insulating disk 810 may be made of (but is not limited to) a semi-rigid foam material that provides spring-like resistance to downward forces and may have a thermal conductivity (κ) in the range of 0.02 to 0.15 W / (m·K). In various embodiments, the semi-rigid foam material may be a polyethylene foam blend, an alternative polymer foam, or a stack of foam materials. The insulating disk 810 includes a recess 812 in its upper surface that receives a contact disk 815 made of a thermally conductive material. In some embodiments, the contact plate 815 may be made of (but is not limited to) copper, copper alloy, silver, silver alloy, aluminum, or aluminum alloy. In some aspects, the contact plate 815 has a thermal conductivity greater than 150 watts / (m·K) (κ > 150 W / (m·K)). In some embodiments, the contact plate 815 may be plated with a coating such as nickel or gold to prevent corrosion of the contact plate 815. A thermocouple connector 820 is adhered to the underside of the contact plate 815, typically at the center of the contact plate 815. In some aspects, the thermocouple connector 820 may be attached to the contact plate 815 via a solder contact 825. In some embodiments, a resistance temperature detector may be used instead of the thermocouple connector. Thermocouple leads 830 exit the underside of the insulating plate 810 through a channel 835. In some aspects, the contact plate 815 may be bonded to the insulating plate 810 via adhesive contacts.

[0069] In operation, the frozen bag or sample container can be considered a heat sink load. The heat sink load, placed on the lower heater plate 230, contacts the contact pad 815, thereby generating a dynamic heat flow through the sensor system comprising the lower heater plate 230, the insulating pad 810, the contact pad 815, and the frozen bag container. It should be understood that each of the one or more thermal sensors in the lower heater plate 230 (e.g., temperature sensor island 675) can be configured as follows: Figure 4Such sensor systems can be controlled individually or in combination based on a set of measured sensor data. The insulating disk 810 can be formed of a material selected to have the lowest thermal conductivity for the sensor system, and therefore, under the conditions of the temperature flux established between the lower heater plate 230 and the freezer bag container, the maximum temperature drop in the thermal path will occur across the insulating disk 810. Therefore, the temperature of the contact disk 815 will be closely coupled to the temperature of the freezer bag. In other words, the temperature measurement will be relatively more specific for the section of the freezer bag within the lower heater plate 230 corresponding to the thermal sensor.

[0070] As the temperature of the frozen bag container, placed in contact with the lower heater plate 230, begins to rise rapidly, the temperature difference between the lower heater plate 230 and the frozen bag container decreases, and the amplitude of the heat flux through each sensor system continuously changes, exhibiting localized variations as measured at each thermal sensor in the lower heater plate 230. Therefore, the temperature of the contact plate 815 does not necessarily need to reach equilibrium with the frozen bag container; rather, the temperature of the contact plate 815 becomes a relative substitute for the temperature of the frozen bag container in the region of the interface between the contact bag and the contact plate 815. Because the temperature of the frozen bag container increases and the temperature difference between the lower heater plate 230 and the frozen bag container decreases, the temperature of the sensor contact plate 815 will more closely represent the temperature of the frozen bag container reaching a point where the phase transition of the contents of the frozen bag container is nearing completion. The temperature of the contact plate is correlated with the temperature recorded by sensors mounted on the inner wall of the bag, with an accuracy of ±10%.

[0071] Therefore, the temperature of the sensor contact plate 815 can be used as a precise and repeatable measure of the completion state of the phase change of the contents of the frozen bag container. Thus, the temperature profile derived from the sensor contact plate 815 can serve as the primary or exclusive data stream for the thawing algorithm in the control instrument, completing the thawing sequence. Applications with multiple sensors in contact with the frozen bag container (e.g.) Figure 6 (As shown) allows the temperature profile of the freezer bag container to be measured at different locations on the container and integrated into a more complex data processing algorithm. The data processing algorithm can compensate for temperature gradients within the container or heater plate and also provides redundant sensors to ensure functionality in the event of failure of one of the sensors.

[0072] Different uses of the defrosting device should be understood from the figures above, including but not limited to the exemplary operating sequences described herein. It should be recognized that the event sequences described below represent one of many possible specific event sequences that can be applied to embodiments of the invention, and are not intended to limit in any way the states, phases, or event sequences that may be associated with the use of the instrument.

[0073] IV. Computer Systems

[0074] Multiple embodiments of this technology may include a defrosting system. The defrosting system may include defrosting devices, which may be any defrosting devices described herein. The defrosting system may also include a computer system, which includes instructions, when executed, to control the defrosting devices to perform methods for defrosting.

[0075] Any computer system mentioned in this article can utilize any suitable number of subsystems. Examples of such subsystems are... Figure 6 The computer system is illustrated in computer system 10. In some embodiments, the computer system includes a single computer device, wherein a subsystem may be a component of the computer device. In other embodiments, the computer system may include multiple computer devices, each of which is a subsystem having internal components. The computer system may include desktop and laptop computers, tablets, mobile phones, other mobile devices, and cloud-based systems.

[0076] Figure 6 The subsystems shown are interconnected via system bus 75. Additional subsystems are shown, such as printer 74, keyboard 78, storage device 79, monitor 76 (e.g., display screen, such as LED) coupled to display adapter 82, etc. External devices and input / output (I / O) devices coupled to I / O controller 71 can be connected to the computer system via any number of devices known in the art, such as input / output (I / O) port 77 (e.g., USB). For example, I / O port 77 or external interface 81 (e.g., Ethernet, Wi-Fi, etc.) can be used to connect computer system 10 to a wide area network, such as the Internet, a mouse input device, or a scanner. The interconnection via system bus 75 allows central processing unit 73 to communicate with each subsystem and control the execution of multiple instructions from system memory 72 or storage device 79 (e.g., a fixed disk, such as a hard disk or optical disk), as well as the exchange of information between subsystems. System memory 72 and / or storage device 79 can embody a computer-readable medium. Another subsystem is a data collection device 85, such as a camera, microphone, accelerometer, etc. Any data mentioned in this article can be output from one component to another, and can also be output to the user.

[0077] A computer system may include multiple identical components or subsystems, for example, connected together via an external interface 81, an internal interface, or via a removable storage device that can be connected and removed from one component to another. In some embodiments, the computer system, subsystem, or device may communicate via a network. In such a case, one computer may be considered a client, and another computer may be considered a server, where each may be part of the same computer system. The client and server may each include multiple systems, subsystems, or components.

[0078] Aspects of the embodiments may be implemented in a modular or integrated manner as control logic using hardware circuitry (e.g., application-specific integrated circuits or field-programmable gate arrays) and / or computer software having a typically programmable processor. As used herein, the processor may include a single-core processor, a multi-core processor on the same integrated chip, multiple processing units on a single circuit board, or networked and dedicated hardware. Based on the disclosure and teachings provided herein, those skilled in the art will recognize and understand other ways and / or methods of implementing embodiments of the invention using hardware and combinations of hardware and software.

[0079] Any software component or function described in this application may be implemented as software code, which will be executed by a processor using any suitable computer language (such as, for example, Java, C, C++, C#, Objective-C, Swift) or a scripting language (such as Perl or Python) using, for example, conventional or object-oriented techniques. The software code may be stored as a series of instructions or commands on a computer-readable medium for storage and / or transmission. Suitable non-transitory computer-readable media may include random access memory (RAM), read-only memory (ROM), magnetic media (such as hard disk drives or floppy disks), or optical media (such as optical discs (CDs) or DVDs (Digital Universal Discs) or Blu-ray discs), flash memory, etc. The computer-readable medium may be any combination of such storage or transmission devices.

[0080] Such programs can also be encoded and transmitted using carrier signals adapted for transmission over wired, optical, and / or wireless networks (including the Internet) conforming to various protocols. Thus, computer-readable media can be created using data signals encoded with such programs. Computer-readable media encoded with program code can be packaged with compatible devices or provided separately from other devices (e.g., downloaded via the Internet). Any such computer-readable medium can reside on or within a single computer product (e.g., a hard drive, CD, or an entire computer system) and can exist on or within different computer products within a system or network. A computer system may include a monitor, printer, or other suitable display for providing a user with any of the results mentioned herein.

[0081] Any method described herein can be performed, wholly or partially, by a computer system including one or more processors configured to perform the steps. Therefore, embodiments may relate to computer systems configured to perform the steps of any method described herein, potentially utilizing different components that perform corresponding steps or groups of steps. Although presented as numbered steps, the steps of the methods herein may be performed at the same time or at different times or in a different order. Furthermore, portions of these steps may be used in conjunction with portions of other steps of other methods. Moreover, all or part of a step may be optional. Additionally, any step in any method may be performed using modules, units, circuits, or other devices of a system for performing those steps.

[0082] The specific details of particular embodiments may be combined in any suitable manner without departing from the spirit and scope of the embodiments of the invention. However, other embodiments of the invention may relate to specific embodiments associated with each individual aspect or a particular combination of these individual aspects.

[0083] The above description of exemplary embodiments of the present disclosure has been presented for purposes of illustration and description. It is not intended to be exhaustive or to limit the present disclosure to the precise forms described, and many modifications and variations are possible in light of the above teachings.

[0084] The use of “a,” “an,” or “the” is intended to mean “one or more” unless explicitly stated otherwise. The use of “or” is intended to mean “including or,” not “excluding or,” unless explicitly stated otherwise. Referring to the “first” component does not necessarily require providing the second component. Furthermore, referring to the “first” or “second” component does not limit the component to a specific location unless explicitly stated otherwise. The term “based on” is intended to mean “at least partially based on.”

[0085] The specific details of particular embodiments may be combined in any suitable manner without departing from the spirit and scope of the embodiments of the invention. However, other embodiments of the invention may relate to specific embodiments associated with each individual aspect or a particular combination of these individual aspects.

[0086] For purposes of illustration and description, the foregoing description of exemplary embodiments of the invention has been presented. It is not intended to be exhaustive or to limit the invention to the precise forms described, and many modifications and variations are possible in light of the foregoing teachings.

[0087] In the above description, numerous details have been set forth for illustrative purposes to provide an understanding of different embodiments of the present technology. However, it will be apparent to those skilled in the art that certain embodiments may be practiced without some of these details or with additional details.

[0088] Several embodiments have been described, and those skilled in the art will recognize that various modifications, alternative constructions, and equivalents can be used without departing from the spirit of the invention. Furthermore, many well-known processes and elements have not been described to avoid unnecessarily obscuring the invention. Moreover, details of any particular embodiment may not always be present in variations of the embodiments, or may be added to other embodiments.

[0089] When providing a range of values, it should be understood that, unless the context explicitly specifies otherwise, each intermediate value between the upper and lower limits of the range is also specifically disclosed to be one-tenth of the lower limit unit. This covers every smaller range between any stated value or intermediate value in the range and any other stated value or intermediate value in the range. The upper and lower limits of these smaller ranges may be independently included or excluded from the range, and each range in which any, and no, one or both limitations are included is also included in the invention, subject to any specifically excluded limitations within the range. Where a stated range includes one or both of these limitations, ranges excluding one or both of those included limitations are also included.

[0090] As used herein and in the appended claims, the singular forms “a,” “an,” or “the” include plural indicators unless the context clearly specifies otherwise. Thus, for example, reference to “a method” includes multiple such methods, and reference to “the heater” includes reference to one or more heaters and their equivalents known to those skilled in the art, etc. For purposes of clarity and understanding, the invention has now been described in detail. However, it should be understood that certain changes and modifications may be practiced within the scope of the appended claims.

[0091] For all purposes, all patents, patent applications, publications, and descriptions mentioned herein are incorporated herein by reference in their entirety. None are acknowledged as prior art.

Claims

1. A method for thawing frozen samples in a bagged container, the method comprising: A plurality of first temperatures are measured in the bag container in contact with the first and second surfaces, each of the plurality of first temperatures being measured by a different sensor of a plurality of sensors, each of the plurality of sensors being configured to measure the temperature at a different location in the bag container; Each of the plurality of first temperatures is compared with a first threshold. The comparison is used to determine a subset of the plurality of sensors, wherein each sensor in the subset of the plurality of sensors measures a first temperature below the first threshold. The frozen sample is heated using a first heater array and simultaneously heated using a second heater array. Multiple second temperatures of the bag container are measured using a subset of the multiple sensors; When one of the plurality of second temperatures exceeds a second threshold, the second threshold indicates that the partially thawed sample is in the bag container: The partially thawed sample was heated using the first heater pack, and The heating of the partially thawed sample using the second heater array is terminated; and the heating of the partially thawed sample using the first heater array is terminated after a certain period of time has been spent heating the partially thawed sample using the first heater array.

2. The method according to claim 1, wherein, When heating of the partially thawed sample using the first heater is stopped, the majority of the partially thawed sample is an aqueous solution, and the solid phase remains in the partially thawed sample.

3. The method according to claim 2, wherein: Heating of the partially thawed sample was terminated at a predetermined time, and The predetermined time is calculated using empirically derived phase transition duration.

4. The method of claim 2, further comprising: Multiple third temperatures of the bag container are measured using a subset of the multiple sensors. The termination of heating of the partially thawed sample using the first heater array is performed when the third temperature among the plurality of third temperatures exceeds a third threshold.

5. The method according to claim 4, wherein, The third threshold is determined based on experience.

6. The method of claim 1, further comprising: Multiple third temperatures of the bag container are measured using a subset of the multiple sensors, and The plurality of third temperatures are compared with the reference temperature curve. The termination of heating of the partially thawed sample using the first heater array is performed when the third temperature among the plurality of third temperatures deviates significantly from the reference temperature curve.

7. The method of claim 1, further comprising: The bag-type container is configured to contact the first surface and the second surface such that: The port of the bag-type container is adjacent to the first end of the first surface and the first end of the second surface; and The port of the bag-type container is positioned closer to the first heater row than the second heater row.

8. The method of claim 1, further comprising: The bag-type container is configured such that: The bag-shaped container overlaps with the first surface at a first portion of the first surface and does not overlap with the first surface at a second portion of the first surface; as well as The bag-shaped container overlaps with the second surface at a first portion and does not overlap with the second surface at a second portion. The first heater array is positioned closer to the first portion of the first surface and the first portion of the second surface than the second portion of the first surface and the second portion of the second surface. The second heater array is positioned closer to the second portion of the first surface and the second surface than the first portion of the first surface and the first portion of the second surface. The bag-type container has a smaller surface area than the first surface. The surface area of ​​the bag-type container is smaller than that of the second surface.

9. The method according to claim 8, wherein, The surface area of ​​the bag-type container is 30% or less of the surface area of ​​the first surface.

10. The method of claim 1, further comprising: The bag-shaped container is configured such that a line perpendicular to the first surface extends through the second heater row but does not extend through the bag-shaped container.

11. The method of claim 1, further comprising: Receive user input specifying the size of the bag-type container; as well as Sensors are excluded from a subset of the plurality of sensors based on the size of the bag-type container.

12. The method of claim 11, further comprising: Receive user input specifying the type of media in the bag container.

13. The method according to claim 1, wherein: Measuring a plurality of first temperatures of the bag-type container in contact with the first and second surfaces includes measuring the plurality of first temperatures at five locations that are not uniformly spaced on the first and second surfaces.

14. The method according to claim 1, wherein, The subset of the plurality of sensors includes fewer sensors than the plurality of sensors.

15. The method according to claim 1, wherein: The frozen sample is the first frozen sample, and The bag-type container is a first bag-type container characterized by a first size. The method further includes: Thawing a second frozen sample in a second bag container that is in contact with the first surface and the second surface, wherein the second bag container is characterized by a second size different from the first size.

16. The method according to claim 1, wherein, The first threshold is lower than the temperature of the bag container as measured when the bag container initially comes into contact with the first surface and the second surface.

17. The method according to claim 1, wherein: Heating the frozen sample using the first heater array includes setting a temperature setpoint for the first heater array within the range of 37°C to 45°C, and Heating the frozen sample using the second heater array includes setting a temperature setpoint for the second heater array.

18. The method according to claim 1, wherein, Heating the partially thawed sample using the first heater array includes setting a temperature setpoint for the first heater array within the range of 37°C to 45°C.

19. The method of claim 1, further comprising: Bring the bag-type container into contact with the first surface. Make the bag-shaped container contact the second surface, and The bag-shaped container is sandwiched between the first surface and the second surface.

20. The method of claim 19, further comprising: Load the bagged container into the drawer of the device, and After terminating heating of the partially thawed sample using the first heater array, the bag container is removed from the device.

21. A defrosting system, comprising: The thawing device includes: First surface, Second surface, Multiple sensors, The first heater row, and The second heater row; and A computer system, the computer system including instructions that, when executed, control the defrosting device to perform the method as described in claim 1.

22. The defrosting system according to claim 21, wherein, The first surface is a first plate, and the second surface is a second plate.

23. The defrosting system according to claim 21, wherein, The multiple sensors are unevenly distributed on the first surface and the second surface.

24. The defrosting system according to claim 21, wherein, Compared to the second heater array being disposed at the first end of the first surface and the first end of the second surface, the first heater array is disposed closer to the first end of the first surface and the first end of the second surface.

25. The defrosting system according to claim 21, wherein, When the bag containing the frozen sample is placed between the first surface and the second surface for thawing, a line perpendicular to the first surface extends through the second heater row but does not extend through the bag.

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