Incubator system for assisted reproductive technology

Abstract

The present disclosure relates to a method of controlling an incubator including controlling an environmental condition of an incubation chamber for a first time period, wherein a pH value of media surrounding a biological specimen in a culture dish in the incubation chamber is affected by the environmental condition. The method includes controlling the environmental condition of the incubation chamber for a second time period, wherein the pH value of media surrounding the biological specimen in the culture dish during the second time period is different than the pH value during the first time period.

Description

INCUBATOR SYSTEM FOR ASSISTED REPRODUCTIVE TECHNOLOGYCLAIM OF PRIORITY

[0001] This application claims priority to U.S. Patent Application No. 63 / 613,853 filed on December 22, 2023, the entire contents of which are hereby incorporated by reference.TECHNICAL FIELD

[0002] The present disclosure relates to assisted reproductive technology, and particularly, an incubator for assisted reproductive technology.BACKGROUND

[0003] Assisted reproductive technology (ART) protocols involve maturing a fertilized oocyte into an embryo within a culture dish that resides in an incubator. During the protocol, the incubator maintains ideal temperature and atmospheric conditions for the embryo and media (z.e., the drop) that overlays the embryo within the culture dish so that the pH and temperature of the drop remain constant. Different types of media provide nutrients that the embryo needs at each stage of embryo growth: fertilization media, cleavage media, and blastocyst media. After a growth stage is complete, the embryo is moved to a different media drop and sometimes to a different incubator with differing gas concentrations.

[0004] Moving embryos between media drops generally takes place under a microscope. For example, each embryo is picked up in a pipette from a dish, moved into a wash drop containing the destination media of another dish where it is injected and aspirated several times to wash the old media off the embryo, and then injected into a drop of the new media. The dish is then placed into, or returned to, the incubator. This process is typically performed between the fertilization stage and the cleavage stage (on days zero or one of an embryos development, for example), and repeated between the cleavage stage and the blastocyst stage (day three, of an embryos development, for example).

[0005] To facilitate an undisturbed culture, a “single step” media was developed by combining aspects of cleavage stage and blastocyst stage media to enable the embryo to develop from a fertilized egg to a mature embryo in a single media drop. This single stage media has a pH value that is acceptable for both cleavage and blastocyst stage, but not optimal for both. Furthermore, current incubators have a single gas setting, and all incubation chambers of that incubator are set at the single gas setting.SUMMARY

[0006] The present disclosure relates to an incubator having a customizable gas management system that provides a controlled environment suitable for storing and culturing human gametes and embryos at various stages of gamete and embryo development.

[0007] In a first example aspect, a method of controlling an incubator may include controlling an environmental condition of an incubation chamber for a first time period, wherein a pH value of media surrounding a biological specimen in a culture dish in the incubation chamber is affected by the environmental condition. The method may include controlling the environmental condition of the incubation chamber for a second time period, wherein the pH value of the media surrounding a biological specimen in the culture dish during the second time period is different than the pH value of media during the first time period.

[0008] In a second example aspect, a method of incubating one or more embryos may include detecting a first gas concentration in a gas management system of an incubator using one or more of a Carbon dioxide gas (CO2) sensor and an O2 sensor. The method may include comparing a first detected gas concentration with a first stage O2 or CO2 concentration suitable for a first embryonic growth stage, and delivering one or more of a Nitrogen gas (N2) and CO2 to an interior volume of an incubation chamber of the incubator to provide the first stage O2 or CO2 concentration in the interior volume of the incubation chamber. The method may include detecting a second gas concentration in the gas management system using the one or more of the CO2 sensor and the O2 sensor, and comparing a second detected gas concentration with a second stage O2 or CO2 concentration suitable for a second embryonic growth stage. The method may include delivering the one or more of the N2 and the CO2 to the interior volume of the incubation chamber to provide the second stage O2 or CO2 concentration in the interior volume of the incubation chamber.

[0009] In a third example aspect, an incubator system for assisted reproductive technologies may include a nitrogen gas (N2) source, a carbon dioxide gas (CO2) source, a gas feed line fluidly coupleable with one or more of the N2 source and the CO2 source, and an incubation chamber operably coupleable to the gas feed line. The incubation chamber may include an inlet, an outlet, and an interior volume disposed between the inlet and the outlet. The interior volume may be sized to receive a dish holding a biological specimen. An inlet valve may be operably coupleable to the inlet of the incubation chamber. The inlet valve may be movable to an open position in which the gas feed line is in fluid communication with the interior volume. An outlet valve may be operably coupleable to the outlet of theincubation chamber, the outlet valve movable to an open position to remove a gas mixture from the interior volume. A pH value of media surrounding a biological specimen disposed in the interior volume of the incubation chamber may be controllable by operating one or more of the inlet valve and the outlet valve.

[0010] In a fourth example aspect, an incubator system for assisted reproductive technologies may include a Nitrogen gas (N2) source, a Carbon dioxide gas (CO2) source, and a gas mixing chamber fluidly coupled to the N2 source and the CO2 source. The gas mixing chamber may include an inlet and an outlet. A gas feed line may be coupled to the outlet of the gas mixing chamber, and a gas recirculation line may be coupled to the inlet of the gas mixing chamber. An incubation chamber may be fluidly coupled to the gas mixing chamber. The incubation chamber may include an inlet, an outlet, and an interior volume disposed between the inlet and the outlet. The interior volume may be sized to receive a dish holding a biological specimen. An inlet valve may be operably coupled to the inlet of the incubation chamber. The inlet valve may be movable to an open position in which the gas feed line is in fluid communication with the interior volume of the incubation chamber, and to a different position in which the gas feed line is not in fluid communication with the interior volume. A pH value of media surrounding a biological specimen disposed in the interior volume of the incubation chamber may be controllable by operating the inlet valve.

[0011] In a fifth example aspect, an incubator system for assisted reproductive technologies may include a Nitrogen gas (N2) source, a Carbon dioxide gas (CO2) source, and a first gas mixing chamber fluidly coupled to the N2 source and the CO2 source and configured for containing a first gas mix having a CO2 gas concentration in a range of 4% CO2 to 7% CO2 and an oxygen gas (O2) concentration in a range of 4% O2 to 6% O2. A second gas mixing chamber may be fluidly coupled to the N2 source and the CO2 source and configured for containing a second gas mix having a CO2 gas concentration in a range of 4% CO2 to 7% CO2 and an O2 concentration in a range of 1% O2 to 4% O2. An incubation chamber may be fluidly coupled to the first gas mixing chamber and the second gas mixing chamber. The incubation chamber may include an inlet, an outlet, and an interior volume disposed between the inlet and the outlet. The interior volume may be configured for receiving a dish holding a biological specimen. A first gas feed line may couple the first gas mixing chamber and the inlet. A second gas feed line may couple the second gas mixing chamber and the inlet. An O2 concentration of the interior volume of the incubation chamber may be controllable by delivering one or more of the first gas mix and the second gas mix through the inlet of the incubation chamber.

[0012] In a sixth example aspect, an incubator system for assisted reproductive technologies may include a first premixed gas source comprising Oxygen gas (O2), Nitrogen gas (N2), and Carbon dioxide gas (CO2). A second premixed gas source may include O2, N2, and CO2. A first gas feed line may be coupled to the first premixed gas source, and a second gas feed line may be coupled to the second premixed gas source. A gas recirculation line may be coupled to the first premixed gas source. An incubation chamber may be fluidly coupled to the first premixed gas source, and may include an inlet, an outlet, and an interior volume disposed between the inlet and the outlet. The interior volume may be sized to receive a dish holding a biological specimen. An inlet valve may be operably coupled to the inlet of the incubation chamber. The inlet valve may be movable to an open position in which the first gas feed line is in fluid communication with the first premixed gas source, and to a different position in which the first gas feed line is not in fluid communication with the first premixed gas source. A pH value of media surrounding a biological specimen disposed in the interior volume of the incubation chamber is controllable by operating the inlet valve. The inlet valve may be movable to a second open position, in which the second gas feed line is in fluid communication with the interior volume of the incubation chamber, and the first gas feed line may be fluidly isolated from the first premixed gas source.

[0013] In accordance with any one of the first, second, third, fourth, fifth, and sixth aspects, the method of controlling an incubator, the method of incubating one or more embryos, and the incubator system for assisted reproductive technologies, may include any one of the following forms.

[0014] In one example, controlling the environmental condition for the first time period may include delivering a gas mixture having a CO2 concentration in a range of 4% to 7.0%.

[0015] In another example, controlling the environmental condition for the first time period may include delivering the gas mixture having a CO2 concentration in a range of 4.9% to 5.1%.

[0016] In some examples, controlling the environmental condition for the second time period may include delivering a gas mixture having a CO2 concentration in a range of 3% to 6%.

[0017] In some examples, controlling the environmental condition for the second time period may include delivering the gas mixture having a CO2 concentration in a range of 3.8% to 4.2%.

[0018] In other examples, controlling the environmental condition for the first time period may include delivering a gas mixture of N2 and CO2 for day one and day two of an embryonic growth cycle.

[0019] In one form, the method may include premixing the gas mixture in a gas mixing chamber.

[0020] In another form, controlling the environmental condition for the second time period may include delivering a different gas mixture of N2 and CO2 for day three, day four, and day five of the five to seven-day embryonic growth cycle.

[0021] In some forms, the method may include premixing the different gas mixture in a different gas mixing chamber.

[0022] In other forms, controlling the environmental condition for the first time period may include setting an internal pressure of the incubation chamber in a range of 111 kPa to 132 kPa.

[0023] In some forms, controlling the environmental condition for the first time period may include opening an inlet valve operably coupled to the incubation chamber, and closing an outlet valve operably coupled to the incubation chamber.

[0024] In one aspect, controlling the environmental condition for the second time period may include setting an internal pressure of the incubation chamber in a range of 101 kPa to 111 kPa.

[0025] In another aspect, controlling the environmental condition for the second time period may include opening the outlet valve.

[0026] In some aspects, controlling the environmental condition for the first time period may include increasing an internal pressure of the incubation chamber from about 101 kPa.

[0027] In other aspects, controlling the environmental condition for the second time period may include decreasing the internal pressure of the incubation chamber to a different predetermined pressure.

[0028] In one example, controlling the environmental condition for the first time period may include maintaining an internal pressure in a range of 110 kPa to 130 kPa, and controlling the environmental condition for the second time period comprises decreasing the internal pressure to 101 kPa.

[0029] In another example, controlling the environmental condition for the first time period may include varying an internal pressure of the incubation chamber by delivering one or more of a high pressure N2 and a high pressure CO2.

[0030] In some examples, controlling the environmental condition for the second time period may include varying the internal pressure of the incubation chamber by delivering the one or more of a high pressure N2 and a high pressure CO2.

[0031] In some examples, the method may include controlling an environmental condition of a second incubation chamber for a first time period, wherein a pH value of media surrounding a biological specimen in a culture dish in the second incubation chamber is affected by the environmental condition.

[0032] In one example, the biological specimen in the second incubation chamber is at a different growth stage than a growth stage of the biological specimen in the first incubation chamber.

[0033] In one example, the method may include controlling the environmental condition of the second incubation chamber for a second time period, wherein the pH value of media surrounding the biological specimen in the culture dish in the second incubation chamber during the second time period is different than the pH value of media in the second incubation chamber during the first time period.

[0034] In other examples, comparing the first detected gas concentration may include comparing a first detected O2 concentration with an O2 concentration in a range of 4% to 6%. In one form,

[0035] In another form, comparing the second detected gas concentration may include comparing a second detected O2 concentration with an O2 concentration in a range of 4.5% to 5.5%.

[0036] In some forms, the method may include detecting a first CO2 concentration in the gas management system using the CO2 sensor.

[0037] In one form, the method may include comparing a first detected CO2 concentration with a CO2 concentration in a range of 4% to 7%.

[0038] In other forms, the method may include detecting a second CO2 concentration in the gas management system using the CO2 sensor.

[0039] In one form, the method may include comparing a second detected CO2 concentration with a CO2 concentration in a range of 3% to 6%.

[0040] In some forms, detecting the first gas concentration of the gas management system may include detecting the first gas concentration of a first gas mixing chamber fluidly coupled to the incubation chamber.

[0041] In one aspect, detecting the second gas concentration of the gas management system may include detecting the second gas concentration of a second gas mixing chamber fluidly coupled to the incubation chamber.

[0042] In another aspect, detecting one or more of the first and second gas concentrations of the gas management system may include detecting the one or more of the first and second gas concentrations of the interior volume of the incubation chamber.

[0043] In some aspects, delivering the one or more of the N2 and the CO2 to provide the first stage O2 or CO2 concentration may include delivering the one or more of the N2 and the CO2 for days zero, one, and two of a five to seven-day embryonic growth cycle.

[0044] In one aspect, the first stage O2 or CO2 concentration may include a CO2 concentration of about 5%.

[0045] In other aspects, delivering the one or more of the N2 and the CO2 to provide the second stage O2 or CO2 concentration may include delivering the one or more of the N2 and the CO2 for days three onward of the five to seven-day embryonic growth cycle.

[0046] In one aspect, the second stage O2 or CO2 concentration may include a CO2 concentration of about 4%.

[0047] In one example, delivering the one or more of the N2 and the CO2 to provide the first stage O2 or CO2 concentration may include delivering a gas mixture of N2 and CO2 from a first gas mixing chamber.

[0048] In another example, delivering the one or more of the N2 and the CO2 to provide the second stage O2 or CO2 concentration may include delivering a different gas mixture from a second gas mixing chamber.

[0049] In some examples, the method may include detecting a pressure value of the interior volume using a pressure sensor.

[0050] In some examples, the method may include comparing a detected pressure value with a first stage pressure value in a range of 1.1 atm to 1.3 atm.

[0051] In one example, the first stage pressure value may be suitable for the first embryonic growth stage.

[0052] In other examples, the method may include modifying a pressure of the interior volume during days zero, one, and two of a five to seven-day embryonic growth cycle to provide the interior volume with the first stage pressure value.

[0053] In one form, the method may include detecting a second pressure value of the interior volume using the pressure sensor, and comparing a second detected pressure value with a second stage pressure value in a range of 0.9 atm to 1.1 atm.

[0054] In one form, the second stage pressure value may be suitable for the second embryonic growth stage.

[0055] In another form, the method may include modifying the pressure of the interior volume during days three onward, of the five to seven-day embryonic growth cycle to provide the interior volume with the second stage pressure value.

[0056] In some forms, modifying the pressure may include varying the pressure of the incubation chamber.

[0057] In other forms, modifying the pressure may include delivering one or more of a high pressure N2 and a high pressure CO2 to the interior volume of the incubation chamber.

[0058] In some forms, delivering one or more of the high pressure N2 and the high pressure CO2 may include delivering one or more of the high pressure N2 and the high pressure CO2 at more than 120 kPa.

[0059] In one aspect, modifying the pressure may include increasing the pressure of the incubation chamber from 1 atm to 1.2 atm during the first embryonic growth stage.

[0060] In another aspect, the method may include recycling a gas mixture delivered to the incubation chamber to a mixing chamber.

[0061] In some aspects, the method may include recycling a different gas mixture delivered to the incubation chamber to a second gas mixing chamber.

[0062] In other aspects, the steps of detecting, comparing, and delivering may be performed with respect to a second incubation chamber of the incubator.

[0063] In one example, a processor may be configured to control an environmental condition of the incubation chamber for a first time period.

[0064] In one example, a pH value of media surrounding a biological specimen in a culture dish in the incubation chamber may be affected by the environmental condition.

[0065] In another example, the processor may be configured to control the environmental condition of the incubation chamber for a second time period.

[0066] In one example, the pH value of media during the second time period may be different than the pH value of media during the first time period.

[0067] In some examples, a gas mixing chamber may be fluidly coupleable to the N2 source and the CO2 source, the gas feed line fluidly coupleable to the gas mixing chamber.

[0068] In some examples, a second gas feed line and a second gas mixing chamber may be fluidly coupleable to the N2 source and CO2 source and to the second gas feed line.

[0069] In other examples, the inlet valve may be movable to a second open position in which the second gas feed line is in fluid communication with the interior volume of the incubation chamber, and the gas feed line is fluidly isolated from the mixing chamber.

[0070] In one form, when the outlet valve is in the open position, the interior volume may be in fluid communication with the gas mixing chamber.

[0071] In another form, the outlet valve may be movable to a second open position in which the interior volume is in fluid communication with the second gas mixing chamber.

[0072] In some forms, the gas mixing chamber may have a CO2 gas concentration in a range of 4% CO2 to 7% CO2, and an O2 concentration in a range of 4% O2 to 6% O2.

[0073] In other forms, the second gas mixing chamber may have a CO2 gas concentration in a range of 3% CO2 to 6% CO2 and an O2 concentration in a range of 4% O2 to 6% O2.

[0074] In some forms, one or more of a CO2 sensor and an O2 sensor may be coupled to the gas mixing chamber.

[0075] In one aspect, one or more of a CO2 sensor and an O2 sensor may be coupled to the second gas mixing chamber.

[0076] In another aspect, one or more of a CO2 sensor and an O2 sensor may be coupled to the interior volume of the incubation chamber.

[0077] In some aspects, a pressure sensor may be coupled to the interior volume of the incubation chamber.

[0078] In other aspects, the incubation chamber may include a lid and a locking mechanism coupled to the lid.

[0079] In one aspect, the locking mechanism may be configured to seal the interior volume from atmosphere in a locked configuration.

[0080] In one example, the incubation chamber may include a second inlet operably coupled to a second inlet valve.

[0081] In another example, a second gas feed line may be operably coupled to the second inlet valve and in fluid communication with the N2 source.

[0082] In one example, the gas feed line may be in fluid communication with the CO2 source and operably coupled to the inlet valve.

[0083] In some examples, the system may include a plurality of incubation chambers.

[0084] In one example, an environmental condition of each of the plurality of incubation chambers may be independently controllable.

[0085] In some examples, an outlet valve may be operably coupled to the outlet of the incubation chamber.

[0086] In one example, the outlet valve may be movable to an open position in which the gas recirculation line is in fluid communication with the interior volume, and a different position in which the gas recirculation line is not in fluid communication with the interior volume.

[0087] In other examples, a gas recirculation line may couple the outlet of the incubation chamber to the first gas mixing chamber.

[0088] In one form, a second gas recirculation line may couple the outlet of the incubation chamber to the second gas mixing chamber.

[0089] In another form, the system may include plurality of incubation chambers, wherein an O2 concentration of each of the plurality of incubation chambers may be separably controllable.

[0090] In some forms, the system may include a plurality of inlet valves and a plurality of incubation chambers.

[0091] In one form, each inlet valve may be operably coupled to an inlet of a different incubation chamber.

[0092] In one form, each of the plurality of inlet valves may be separably controllable.

[0093] In other forms, the system may include a processor configured to control an environmental condition of the incubation chamber for a first time period.

[0094] In another form, the processor may be configured to control the environmental condition of the incubation chamber for a second time period.

[0095] Systems and methods described in the present disclosure can include one or more of the following advantages.

[0096] Incubators constructed in accordance with the present disclosure may allow storing and culturing of human gametes and embryos by providing environmental conditions similar to the human fallopian tube and uterus in terms of temperature, physiological pH, and oxygen levels.

[0097] In accordance with certain methods of the present disclosure, media does not have to be replaced or switched out during or between the growth stages.

[0098] In accordance with some methods of the present disclosure, the embryo growth process is streamlined with fewer disruptions between each growth stage.

[0099] In accordance with certain incubators of the present disclosure, there can be standardization across labs in terms of gas concentration, regardless of elevation of the lab. For example, by controlling an internal pressure of the incubator chambers, the incubatormay not need to provide a CO2 concentration based on elevation (barometric pressure) of the lab location.

[0100] As used herein, the terms “top,” “bottom,” “upper,” “lower,” “above,” and “below” are used to provide a relative relationship between structures. The use of these terms does not indicate or require that a particular structure must be located at a particular location in the apparatus.

[0101] Some examples may be described using the expression “coupled” and “connected” along with their derivatives. For example, some arrangements may be described using the term “coupled” to indicate that two or more elements are in direct physical or electrical contact. The term “coupled,” however, may also mean that two or more elements are not in direct contact with each other, but yet still co-operate or interact with each other. The examples described herein are not limited in this context.

[0102] Other aspects, features, and advantages of the present disclosure will be apparent from the following detailed description, figures, and claims.BRIEF DESCRIPTION OF THE DRAWINGS

[0103] Fig. l is a perspective view of an incubator with an open incubation chamber that has a customizable gas management system for providing a controlled environment suitable for storing and culturing human gametes and embryos at various stages of gamete and embryo development.

[0104] Fig. 2 is an enlarged, front perspective view of the open incubation chamber of Fig. 1, showing two culture dishes disposed in the incubation chamber.

[0105] Fig. 3 is a side, cross-sectional view of an open and empty incubation chamber of the incubator of Fig. 1.

[0106] Fig. 4 is a block diagram of the gas management system of the incubator of Fig. 1.

[0107] Fig. 5 is a top, perspective view of a preparation compartment of the incubator of Fig. 1.

[0108] Fig. 6 is a block diagram illustrating communications among various portions of the incubator of Fig. 1.

[0109] Fig. 7 is a simplified, block diagram of a different gas management system of an incubator.

[0110] Fig. 8 is a side, cross-sectional view of an open and empty incubation chamber of an incubator with the gas management system of Fig. 7.

[0111] Fig. 9 is a simplified, block diagram of another gas management system of an incubator.

[0112] Fig. 10 is an enlarged, front perspective view of an open incubation chamber of an incubator with the gas management system of Fig. 9, showing two culture dishes disposed in the incubation chamber.

[0113] Fig. 11 is a side, cross-sectional view of the incubation chamber of Fig. 10, showing the incubation chamber empty.DETAILED DESCRIPTION

[0114] The present disclosure relates to an incubator having a customizable gas management system that provides a controlled environment suitable for storing and culturing human gametes and embryos at various stages of gamete and embryo development during in vitro fertilization / assisted reproductive technology (ART) treatments. The gas management system is customizable to set and control environmental conditions of each incubation chamber (e.g., the Carbon dioxide (CO2) concentration, oxygen (O2) concentration, internal pressure, temperature) of the incubator, thereby providing a suitable environment at each stage of gamete and embryo development. By customizing the CO2 concentration and / or pressure, for example, the gas management systems described herein can affect and adapt a pH value of the media surrounding a biological specimen at each development stage. This is because CO2 buffered media contains a chemical (z.e., bicarbonate) that changes the media’s pH in response to atmospheric CO2 levels. Additionally, at higher altitudes, CO2 levels are raised to maintain the same pH value, and therefore pressure of the environment can affect pH for any given CO2 level. Further, by customizing the O2 concentration to the particular growth stage, the incubation chamber can more closely mirror the O2 concentration levels of the human body (e.g., the uterus during the cleavage stage, and the fallopian tubes during the blastocyst stage). Controlling these environmental conditions for each chamber depending on the particular growth stage of the culture dish of that chamber may streamline storing and culturing human gametes and embryos with minimal disruption and risk of contamination.

[0115] Fig. 1 illustrates an incubator 100 that is designed to house biological specimens as part of a developmental protocol. Example biological specimens that may be housed within the incubator 100 include reproductive specimens (e.g., human and non-human gametes and embryos) and other biological specimens, such as various types of mammalian cell cultures. For example, a human embryo may be cultured within the incubator 100 as partof an ART protocol. In other examples, biological specimens, more generally, may be cultured within the incubator 100 as part of any number of cell culture protocols.

[0116] The incubator 100 is operable by a user (e.g., a scientist or laboratory technician, such as, for example, an embryologist) to provide optimal growth conditions for biological specimens. In the specific example, the incubator 100 has an environment control system to optimize pH and oxygen concentration at each stage of an embryo’s development.Accordingly, the incubator 100 includes a housing 102, a user interface module 106 that is supported on the housing 102, a control system 101 (shown in Fig. 6) that is located within the housing 102, and a separate monitoring system 198 (shown in Fig. 6).

[0117] The housing 102 includes an incubation compartment 108 that forms a flatbed- style configuration, a base compartment 110 that supports the incubation compartment 108, and a preparation compartment 112 that extends upward from the base compartment 110. The incubation compartment 108 includes an outer peripheral wall 114 and a top wall 116 with multiple (e.g., ten) substantially rectangular (e.g., rectangular-round) openings 118 arranged in two (e.g., front and rear) rows.

[0118] Referring to Figs. 1 and 2, the incubation compartment 108 is sized to accommodate multiple receptacles 120 that are respectively installed to the housing 102 at the openings 118. Accordingly, the receptacles 120 include a bottom wall portion 122 and four side wall portions 124A, 124B, 124C, 124D (shown in Figs. 2 and 3) and have the same cross-sectional shape (e.g., substantially rectangular) as the shape of the openings 118. Each receptacle 120 is sized to receive one or more culture dishes 218 directly.

[0119] Referring to Fig. 2, in some examples, each receptacle 120 is attached (e.g., bolted or otherwise fastened or adhered) to the top wall 116 of the incubator 100 along an edge of the respective opening 118. The receptacle 120 can, for example, be screwed or otherwise fastened to the top wall 116 and / or to other structures within the housing 102 of the incubator 100. The receptacle 120 is attached in a manner such that a top surface of the receptacle 120 is substantially flush with the top wall 116 of the incubation compartment 108.

[0120] The receptacle 120, as noted above, includes the bottom wall portion 122 and the side wall portions 124 A, 124B, 124C, 124D, which cooperate with a lid 126 of the incubator 100 to define an incubation chamber 128.

[0121] Referring to Figs. 1-3, the housing 102 is equipped with multiple lids 126 that respectively cover the multiple receptacles 120. Each receptacle 120 and respective lid 126 together form the incubation chamber 128, and each incubation chamber 128 defines an interior volume 133 sized for receiving the culture dishes 218. The lid 126 is coupled to aconnector block 132 with an internal pin that extends through the connector block 132 and the lid 126. The lid 126 is pivotable with respect to the connector block 132 between an open position that provides access to the incubation chamber 128 and a closed position that substantially isolates the incubation chamber 128 from an ambient environment. After the lid 126 has been closed, the incubator 100 is operable to achieve and substantially maintain programmed (e.g., user-set or pre-set) temperature, gas, and / or pressure conditions within the incubation chamber 128.

[0122] Turning to Figs. 2 and 3, a representative incubation chamber 128 is illustrated. The incubation chamber 128 includes an inlet 103, an outlet 115 (shown in dashed lines in Fig. 2), and an interior volume disposed between the inlet 103 and the outlet 115. The inlet 103 is connected to the side wall portion 124A of the receptacle 120 for delivering a gas mixture to the interior volume 133 of the incubation chamber 128. The inlet 103 is operably coupled to an inlet valve 137 (e.g., a three-way valve) that is operably coupled to a first gas feed line 194A and a second gas feed line 194B (Fig. 3). The outlet 114 is operably coupled to an outlet valve 143 (e.g., a three-way valve) that is operably coupled to a first gas recirculation line 197 A and a second gas recirculation line 197B. Environmental conditions (e.g., gas concentrations, humidity, pressure, etc.) of the interior volume 133 of the incubation chamber 128 are controllable by individually operating the inlet valve 137and the outlet valve 143, as will be described in more detail below with reference to Fig. 4.

[0123] The inlet valve 137 is movable to a first open position in which the first gas feed line 194A is in fluid communication with the interior volume 133 and the second gas feed line 194B is not in fluid communication with (or sealed from) the interior volume 133. The inlet valve 137 is also movable to a second open position in which the second gas feed line 194B is in fluid communication with the interior volume 133 and the first gas feed line 194A is not in fluid communication with (or sealed from) the interior volume 133. The inlet valve 137 is also movable to a closed position in which both gas feed lines 194A, 194B are not in fluid communication with (or sealed from) the interior volume 133. The inlet valve 137 may also be moved to intermediate positions between the first and second open positions and the closed position to allow for throttling and to otherwise control the flow of gas (e.g, pressure, flow rate) into the incubation chamber 128. As will be described later in further detail, each of the gas feed lines 194 A, 194B fluidly couple a gas source with the interior volume 133 of the incubator chamber 128 via respective distribution lines 196A, 196B that branch from the gas feed lines 194 A, 194B to each individual chamber 128.

[0124] The outlet or outlet nozzle 115 (shown in dashed lines in Fig. 2) is found in an opposite side wall 124B of the receptacle 120 for removing gas from the interior volume 133 of the incubation chamber 128. The outlet valve 143 is movable to a first open position, in which the interior volume 133 is in fluid communication with a first gas recirculation line 197A and is not in fluid communication with (z.e., sealed closed from) a second gas recirculation line 197B. The outlet valve 143 is also movable to a second open position, in which the interior volume 133 is in fluid communication with the second gas recirculation line 197B and not in fluid communication with (z.e., sealed closed from) the first gas recirculation line 197A. The outlet valve 143 is also movable to a closed position, in which both gas recirculation lines 197 A, 197B are not in fluid communication with (or sealed from) the interior volume 133. The outlet valve 143 may also move to different positions between the first and second open positions and the closed position to allow for throttling and to otherwise control the flow of gas (e.g., concentration, pressure, flow rate) out of the incubation chamber 128. As will be described later in further detail, each of the gas recirculation lines 197A, 197B fluidly couples the interior volume 133 of the incubator chamber 128 with the respective gas source via respective distribution lines 199A, 199B that branch from the gas recirculation lines 194 A, 194B from each individual chamber 128.

[0125] Referring to Figs. 2 and 3, the lid 126 includes a base plate 134 that encloses a broad heating element 136 (e.g., a heater foil) and one or more gaskets 138 that are secured to an inner surface of the base plate 134 (e.g., at one or more respective recesses). The receptacle 120 is also equipped with one or more heating elements 136 (e.g., heater foils) that are exteriorly attached to one or both of the bottom wall portion 122 and the side wall portions 124A, 124B, 124C, 124D within the incubation compartment 108. When the lid 126 is closed against the housing 102, the one or more gaskets 138 seal against the receptacle 120 and the top wall 116 to isolate the incubation chamber 128.

[0126] The receptacle 120 is further equipped with a control temperature sensor 170 (e.g., a thermostat) and a monitoring temperature sensor 172 that are secured exteriorly to the bottom wall portion 122. The control temperature sensor 170 is coupled to the control system 101, and the monitoring sensor 172 may be coupled to a monitoring system 198 that is separate from the incubator 100. The control temperature sensor 170 is also coupled to and controls (e.g., via the control system 101) operation of the heating elements 130, 136 to achieve a set (e.g., target) temperature within the incubation chamber 128. The monitoring sensor 172 monitors a temperature of the receptacle 120 as an indication of the temperaturewithin the incubation chamber 128. The monitoring system 198 is separate (e.g., electrically isolated) from the control system 101.

[0127] The receptacle 120 is made of one or more materials that are suitable for adequately conducting heat generated by the heating elements 136 to an interior region of the incubation chamber 128. Example materials from which the receptacle 120 may be made include aluminum and other thermal and chemically compatible materials. Example materials from which the lid 126 may include materials that provide strong insulation properties, such as plastics, foams or other thermal and chemically compatible materials.

[0128] Returning briefly to Fig. 1, the base compartment 110 of the incubator includes two lateral walls 105, a bottom wall 107, a rear wall 109, and a top wall 111. The top wall 111 opens to the incubation compartment 108 and the preparation compartment 112. The base compartment 110 supports a power connection port on the rear wall 109 for connecting the incubator 100 to a wall outlet via cable connection. The base compartment 110 contains components of a gas management system 180, (optionally) the preparation compartment 112, the control system 101, and other supporting electronic components.

[0129] Referring now to Fig. 4, a gas management system 180 of the incubator 100 is configured to deliver a mixture of nitrogen gas (N2), oxygen gas (O2) and carbon dioxide gas (CO2) to each incubation chamber 128 by way of two gas cycles: a first cycle 181 A filters, delivers, and recycles a first gas mixture, and a second cycle 18 IB filters, delivers, and recycles a second gas mixture. Each cycle 181 A, 181B is coupled to the source of CO2 174 and the source of N2 176. The gas management system 180 includes first and second manifolds 173 A, 173B that are installed to the rear wall 109. Remaining components of the gas management system 180 are contained within the base compartment 110.

[0130] The first manifold 173 A receives and routes CO2 and N2 from the CO2 source 174 and the N2 source 176 to a first gas mixing chamber 178 A through a gas line 182A, and the second manifold 173B receives and routes CO2 and N2 from the CO2 source 174 and the N2 source 176 to a second gas mixing chamber 178B through a gas line 182B. The gas management system 180 includes first and second filtration modules 183 A, 183B along the respective gas lines 182A, 182B, each including a HEPA filter and a volatile organic compound (VOC) filter. Each of the first and second gas mixing chambers 178 A, 178B is equipped with a CO2 sensor 184A, 184B and an oxygen gas (O2) sensor 186A, 186B to respectively monitor CO2 and O2 concentrations of a gas mixture containing the incoming CO2 and N2 gases. Based on concentration measurements acquired at the sensors 184A, 184B, 186A, 186B, one or more valves on the manifolds 173 A, 173B are operated (e.g.,opened and closed) to adjust the flow rates of CO2 and N2 to achieve pre-set (e.g., target) CO2 and O2 concentrations of the gas mixtures. In one example, the first gas mixing chamber 178 A has a gas mixture suitable for a cleavage stage, for example, a concentration of CO2 in a range of approximately 4% (e.g., about 4.25% or more, about 4.5% or more, about 4.75% or more, about 5%) to approximately 7% (e.g., about 6.75% or less, about 6.5% or less, about 6.25% or less, about 6.0% or less, about 5.75% or less, about 5.5% or less, about 5.25% or less, about 5%), and a concentration of O2 in a range of approximately 3% (e.g., about 3.25% or more, about 3.5% or more, about 3.75% or more, about 4% or more, about 4.25% or more, about 4.5% or more, about 4.75% or more, about 5%) to approximately 6% (e.g., about 5.75% or less, about 5.5% or less, about 5.25% or less, about 5.0%), and the second gas mixing chamber 178B has a gas mixture suitable for a blastocyst stage, for example, a concentration of CO2 in a range of approximately 3% (e.g., about 3.25% or more, about 3.5% or more, about 3.75% or more, about 4%) to approximately 6% (e.g., about 5.75% or less, about 5.5% or less, about 5.25% or less, about 5% or less, about 4.75% or less, 4.5% or less, about 4.25% or less, about 4%), and a concentration of O2 in a range of approximately 4% (e.g., about 4.25% or more, about 4.5% or more, about 4.75% or more, about 5%) to approximately 6% (e.g., about 5.75% or less, about 5.5% or less, about 5.25% or less, about 5.0%).

[0131] From the gas mixing chambers 178 A, 178B, the mixed gas flows through gas lines 188 A, 188B to respective gas monitoring chambers 190 A, 190B. Each of the gas monitoring chambers 190A, 190B is located just upstream of the incubation chambers 128 to allow for an additional measurement of the concentration profile of the gas mixture before the mixture is delivered to the incubation chambers 128. In this regard, each of the monitoring chambers 190A, 190B is equipped with an external gas sensor 192A, 192B that measures both CO2 and O2 concentrations. The gas monitoring chambers 190 A, 190B and gas sensors 192A, 192B are operationally separate (e.g., electrically isolated) from the control system 101 to allow for independent verification of the gas concentration profile of the mixture. From the gas monitoring chambers 190A, 190B, the gas mixture flows into respective first and second gas feed lines 194 A, 194B that deliver the gas mixture to the incubation chambers 128 via multiple, respective distribution lines 196A, 196B and respective the gas nozzle 103, or inlet, of each receptacle 120 (Figs. 2 and 3).

[0132] So configured, the gas management system 180 in Fig. 4 is customizable to provide a suitable environment for each of the plurality of incubation chambers 128 of the incubator 100 depending on the stage of development (e.g., cleavage stage and blastocyststage for an embryo) of the biological specimen in the incubation chamber 128. The gas management system 180 can alter the pH value of media surrounding the biological specimen in the culture dish 218 by controlling the CO2 concentration in the interior volume 133 for each of the plurality of incubation chambers 128.

[0133] For example, the first gas mixing chamber 178 A may be set to provide a gas mixture suitable for a cleavage stage (e.g., having a concentration of CO2 of approximately 5%, and a concentration of O2 of approximately 5%). Temperature may be kept constant in a range of approximately 36.5°C to approximately 37.5°C, and pressure may be kept constant at 1.0 atm. Accordingly, the gas management system 180 is arranged to deliver a gas mixture from the first gas mixing chamber 178A to any of the incubation chambers 128 containing an embryo in the cleavage stage, or during days zero, one and two of an embryo’s development (e.g., which could be five to seven days). The second gas mixing chamber 178B is set to provide a gas mixture suitable for a blastocyst stage (e.g., having a concentration of CO2 of approximately 4%, and a concentration of O2 of approximately 5%). Temperature may be kept constant in a range of approximately 36.5°C to approximately 37.5°C, and pressure may be kept at 1.0 atm. Accordingly, the gas management system 180 is arranged to deliver a gas mixture from the second gas mixing chamber 178B to any incubation chamber 128 containing an embryo in the blastocyst stage (or from day 3 onward in an embryo’s development cycle, for example). Put differently, the incubator 100 is configured to simultaneously control environmental conditions of multiple incubation chambers 128 containing biological specimens at various stages of the embryonic cycle. A control system 101 configured to operate the gas management system 180 will be described below in further detail with respect to Fig. 6.

[0134] Additionally, the gas management system 180 is configured to switch gas delivery between the two gas mixing chambers 178 A, 178B based on the growth stage of the biological specimen. To deliver the gas mixture from the first gas mixing chamber 178A into a specific incubation chamber 128, the inlet valve 137 of the specific incubation chamber 128 is movable to a first open position, in which the first gas mixing chamber 178 A is in fluid communication with the interior volume 133 of the incubation chamber 128. To deliver the gas mixture from the second gas mixing chamber 178B to the specific incubation chamber 128, the inlet valve 137 coupled to the specific incubation chamber 128 is movable to a second open position, in which the first gas mixing chamber 178 A is fluidly isolated from the incubation chamber 128, and the second gas mixing chamber 178B is in fluid communication with the interior volume 133 of the incubation chamber 128. Accordingly, when an embryoin the specific incubation chamber 128 transitions from the cleavage stage (days zero, one, and two of a five-seven day embryonic growth cycle, for example) to the blastocyst stage (days three onward, of a five- to seven-day embryonic growth cycle, for example), the inlet valve 137 moves from the first open position to the second open position.

[0135] Referring to Fig. 5, the preparation compartment 112, which may be optionally provided in the incubator 100, provides a space that may be optionally used for a brief period to warm bottles of media and allow them to achieve a gas equilibrium. The preparation compartment 112 includes a cylindrical wall 113, a pivotable lid 117, and a top support wall 119. The top support wall 119 supports an internal receptacle 121 that is sized to accommodate bottles of media. The internal receptacle 121 is equipped with one or more exterior heating elements (not visible) for warming bottles of media and a gas nozzle 123 for delivering a gas mixture from the gas management system 180 to the dish 218 until the dish 218 is ready to be placed within one of the incubation chambers 128. The internal receptacle 121 is also equipped with an exterior temperature sensor (not visible) that monitors a temperature within the compartment 112 and that is coupled to the control system 101. The lid 117 carries an open / close sensor that causes the gas delivery to stop while the lid 117 is open and to flow while the lid is closed.

[0136] Referring to Figs. 1 and 6, the user interface module 106 is a touchscreen display that presents system information to a user. Such information includes settings, monitored properties (e.g., temperatures, gas concentrations, pressure), specimen information (e.g., identification information, development stage, and other information), and visual warnings and alerts, among other information. The module 106 also presents user interfaces that provide input fields for receiving various operational inputs from the user. The user interface module 106 is coupled to the control system 101, as indicated in Fig. 6. In some examples, the user interface may be in wireless communication with a separate computer or user workstation.

[0137] Still referring to Fig. 6, the control system 101 is programmed to control operations of the incubator 100 based on stored data, input parameters received at the user interface module 106, and on sensor data (e.g., temperature sensors, pressure sensors, gas concentration sensors, etc.). Accordingly, the control system 101 is electronically coupled to the incubation chambers 128 and (optionally) preparation compartment 112 (e.g., to the above-discussed heating elements and sensors). The control system 101 is also electronically coupled to the user interface module 106 and to the components of the gas management system 180. The control system 101 includes one or more processors 125 and supportingelectronic components 127 for carrying out its functionalities. The control system 101 includes a wireless transmitter for sending data to a separate computer or workstation. In some examples, the control system 101 may be in wired communication with a separate computer. The control unit 101 also includes a transmitter that sends data (e.g., associated metadata, and other data) wirelessly over a network 258 to one or more computing devices 260 and to a server system 262 to be stored in a database 264.

[0138] For culturing an embryo, an embryologist may input data into the control system 101 each time a new embryo is placed inside an incubator chamber 128. The control system 101 operates a timer based on the development cycle (e.g., a five to seven-day embryonic growth cycle) of the biological specimen placed in the incubator chamber 128. Patient information associated with the embryo and other information can be inputted via the control panel 106 or via a remote workstation 260. During the first and second days of the embryonic growth cycle, the control system 101 is configured to deliver a gas mixture (z.e., the gas mixture having a concentration of CO2 in a range of approximately 4% to approximately 7%, and a concentration of O2 in a range of approximately 4% to approximately 6%) from the first gas mixing chamber 178 A to the incubation chamber 128 by operating one or more control valves 137, 143 coupled to chamber 128. The monitoring system 198 may be coupled to the incubator 100 to continually monitor the gas concentration in the gas mixing chamber 178 A to ensure that the desirable gas mixture is delivered to the incubation chamber 128 holding the embryo in the cleavage stage. At the beginning of the third day of the embryonic growth cycle of the embryo, or once the embryo is seen to meet a specified developmental milestone, the control system 101 operates the control valve 137 to move from the first open position to the second open position to deliver a gas mixture (z.e., the gas mixture having a concentration of CO2 in a range of approximately 3% to approximately 6%, and a concentration of O2 in a range of approximately 4% to approximately 6%) from the second gas mixing chamber 178B to the incubation chamber 128. The monitoring system 198 continually monitors the gas concentration in the second gas mixing chamber 178B to ensure that the desirable gas mixture is delivered to the incubation chamber 128 holding the embryo now in the blastocyst stage.

[0139] The control system 101 is also configured to operate the various heating elements in each incubation chamber based on the temperature sensor data received. For example, the temperature is kept constant at 37°C.

[0140] As indicated in Fig. 6, the monitoring system 198 may be coupled to the gas management system 180 (e.g., to the gas monitoring chambers 190A, 190B, gas sensors192A, 192B, 184A, 184B, 186A, 186B of Fig. 4) and to the incubation chambers 128 (e.g., to the monitoring sensors 172 in Fig. 3). The monitoring system 198 includes one or more processors 129 and supporting electronic components 131 for carrying out its functionalities. Specifically, the monitoring system 198 is configured to receive sensor data of the various gas sensors of the gas management system 180 and temperature sensors of the incubator 100.

[0141] The control system 101, which is in communication with the various sensors, processes the sensor data of the biological specimen in the incubation chamber 128 to further control the environmental conditions of the incubation chamber 128. To affect the pH value of media surrounding the biological specimen for each stage of the embryo’s development, the control system 101 communicates with the control valve 137 to increase CO2 concentration to lower pH or lower CO2 concentration to raise the pH value. The monitoring system 198 is electronically isolated from the control system 101 to provide backup e.g., duplicate or redundant) system checks that cannot be compromised by a state of the control system 101. The monitoring system 198 includes a wireless transmitter for sending data to a separate computer.

[0142] While the gas management system 180 of the incubator of Figs. 1 and 4 has been described having two manifolds 173 A, 173B, two filtration modules 183 A, 183B, two gas mixing chambers 178 A, 178B, two gas lines 188 A, 188B, two gas monitoring chambers 190A, 190B, two gas feed lines 194A, 194B, two gas distribution lines 196A, 196B, and two gas recirculation lines 197A, 197B, in some examples, a gas management system of an incubator may include more than two manifolds, more than two filtration modules, more than two mixing chambers, more than two gas lines, more than two gas monitoring chambers, more than two gas feed lines, more than two gas redistribution lines, and more than two recirculation lines.

[0143] While the incubation chamber 128 of Figs. 1-3 has been described having the inlet 103 and outlet 115 disposed in opposite side wall portions 124A, 124B, in other examples, one or more inlets and one or more outlets may be disposed in the same sidewall portions or different sidewall portions of an incubation chamber.

[0144] While the incubation chamber 128 of Figs. 1-4 has been described having one outlet, in other examples, an incubator may include a plurality of incubation chambers, each including more than one outlet.

[0145] While the gas management system 180 of the incubator of Fig. 4 has been described as having two gas cycles 181 A, 18 IB, in other examples, a gas management system of an incubator may have three gas cycles.

[0146] While the gas management system 180 of the incubator of Fig. 4 has been described as having one outlet valve 143 coupled to two gas recirculation lines 197 A, 197B and one outlet 115, in other examples, gas recirculation may be controlled with two outlet control valves where each outlet valve is coupled to one outlet and one gas recirculation line.

[0147] While the gas management system 180 of the incubator of Fig. 4 has been described as having one inlet valve 137 coupled to two gas feed lines 194 A, 194B and one inlet 103, in other examples, gas delivery may be controlled with two inlet control valves where each control valve is coupled to one inlet and one gas feed line, such as, for example, in the gas management system of Fig. 9.

[0148] While the gas management system 180 of the incubator of Figs. 1 and 4 has been described having two gas cycles 181 A, 18 IB each associated with a manifold, a filtration module, a gas mixing chamber, a gas line, a gas monitoring chamber, a gas feed line, a gas distribution line, and a gas recirculation line, in some examples, a gas management system of an incubator may include only one gas cycle associated with one manifold, one filtration module, one mixing chamber, one gas line, one gas monitoring chamber, one gas feed line, one gas redistribution line, and one recirculation line, as shown in Fig. 7.

[0149] Referring to Fig. 7, for example, a gas management system 380 for an incubator includes one gas cycle having a mixing chamber 378 that dilutes and mixes ambient air with CO2 and N2 from CO2 and N2 sources 374, 376. The gas management system 380 is similar to the gas management system 180 described above and can be installed in the incubator 100 of Fig. 1. Elements of the gas management system 380 in Fig. 7 that are similar to the elements of the gas management system 180 of Fig. 4 are designated by the same reference numeral, incremented by 200. A description of many of these elements is abbreviated or even eliminated in the interest of brevity. The gas management system 380 may also include a manifold, a filtration module, and a gas monitoring chamber, which are not illustrated in Fig. 7, but are similar to those illustrated and described in one of cycles 181 A, 18 IB of Fig.4. The gas management system 380 differs from the gas management system 180 by including one gas mixing chamber 378 and a pressure sensor 472 disposed in each incubation chamber 328.

[0150] While the incubator 100 of Figs. 1 and 5 has been described as having an optional preparation compartment 112, in other examples, the incubator may be constructed without a preparation compartment. In some examples, the incubator may have more incubation chambers, and in other examples, the incubator may have fewer incubation chambers. 1

[0151] While the incubator of 100 of Figs. 1 and 5 has been described as providing an internal pressure of the incubator chambers at approximately 1 atm (about lOlkPa), in other examples, the incubator may control the internal pressure depending on the elevation of the lab. For example, in higher altitude labs, the incubator may be pressurized to 1 atm (e.g., in a range of about l.Oatm to about 1.1 atm) to get the expected results from an environmental condition in the chamber.

[0152] In other examples, such as the example of Figs. 7 and 8, the incubator may control the internal pressure depending on the developmental stage of the embryo. For example, in the cleavage stage, an internal pressure may be in a range of approximately 1.1 atm (e.g., about 1.15 atm or more, about 1.2 atm) to approximately 1.3 atm (e.g., about 1.25 atm or less, about 1.2 atm), and in a blastocyst stage, an internal pressure may be in a range of approximately 0.9 atm (e.g., about 0.95 atm or more, about 1.0 atm) to approximately 1.1 atm (e.g., about 1.05 atm or less, about 1.0 atm).

[0153] While the control system 101 of the incubator of Figs. 1 and 6 has been described as being coupled to temperature sensors in the incubation chambers 128 and gas concentration sensors in the gas management system 180, in other examples, the control system 101 may additionally or alternatively be coupled to pressure sensors and / or gas sensors coupled to the incubation chambers. For example, in Figs. 7 and 8, the control system 101 may be coupled to a pressure sensor 472 of each incubator chamber.

[0154] While the monitoring system 198 of the incubator of Figs. 1 and 6 has been described as being coupled to monitoring temperature sensors 172 of the incubation chambers 128, in other examples, the monitoring system 198 may additionally or alternatively be coupled to pressure sensors and / or gas sensors disposed in the incubation chambers. For example, in Figs. 7 and 8, the monitoring system may be coupled to the pressure sensor 472 coupled to each incubation chamber 328 and CO2 and O2 sensors 384, 386 disposed in the gas mixing chamber 378.

[0155] In Figs. 7 and 8, each incubation chamber 328 includes a pressure sensor 472 coupled to the chamber 328 (e.g., secured interiorly to a bottom wall portion 322, or via a hose or gas line) that monitors an internal pressure of each incubation chamber 328. By controlling the internal pressure of the incubation chamber 328, a control system, such as the control system 101 of Fig. 6, is configured to change the pH value of media surrounding a biological specimen held inside the incubation chamber 328. A control system, such as the control system 101 of Fig. 6, can receive sensor data from the pressure sensors 472, which can then be translated or processed to determine the pH value of the media surroundingbiological specimen. Based on that determination, the control system 101 is configured to control delivery of high-pressure N2 and CO2 (e.g., 130 kPa) from the gas mixing chamber 378 (e.g., by operating the inlet and outlet valves 373 and 343) to provide a suitable pH to the culture dish in the incubation chamber 328 based on the growth stage.

[0156] For example, the control system 101 operates a timer based on the five to seven- day embryonic growth cycle of the embryo placed in each incubator chamber 328. During the first and second days of the embryonic growth cycle of the embryo, the control system 101 is configured to deliver a gas mixture from the gas mixing chamber 378 to the incubation chamber 328. The monitoring system 198 continually monitors the gas concentration in the mixing chamber 378 (via the CO2 and O2 sensors 384, 386), as well as the internal pressure (via the pressure sensor 472) of the incubation chamber 328 holding the embryo in the cleavage stage. The control system 101 receives and processes the sensor data, and operates the inlet and outlet valves 337, 343 coupled to the chamber 328 based on that data. For example, while the embryo is in the cleavage stage, the control system 101 opens the inlet valve 337 coupled to the incubation chamber 328 to deliver a gas mixture having a pressure in a range of approximately 1.1 atm (or about 111 kPa) to approximately 1.3 atm (or about 132 kPa). This may include moving the inlet valve 337 only, the outlet valve 343 only, or moving both the inlet and outlet valves 337, 343. In some examples, the control system 101 may be programmed to open the inlet valve 373 in such a way to slowly ramp up the gas pressure in the chamber 328 during the cleavage stage. In other examples, the control system 101 may be programmed to vary the delivery of a gas mixture to the incubation chamber 328 during the cleavage stage. At the beginning of the third day of the embryonic growth cycle, the control system 101 is configured to operate the control valves 337, 343 to set an internal pressure value in the incubation chamber at a pressure in a range of approximately 0.9 atm (or about 91kPa) to approximately 1.1 atm (or about 11 IkPa). To achieve the desired internal pressure, the control system 101 can move the inlet valve 337 only, the outlet valve 343 only, or move both the inlet and outlet valves 337, 343. In some examples, the control system 101 may be programmed to open the outlet valve 343 to allow the internal gas pressure to drop to atmospheric pressure. In other examples, the control system 101 may be programmed to vary the ventilation of the gas mixture from the incubation chamber 328 during the blastocyst stage.

[0157] In some examples, the gas management system 380 provides high-pressure CO2 and high-pressure N2 at about 130 kPa.

[0158] While the lid 126 of the incubation chamber 128 of Fig. 2 has a gasket 138, in other examples, a lid 326 of an incubation chamber 328 may include a locking mechanism to lock the lid against an incubator housing. For example, in Fig. 8, the incubation chamber 328 includes a locking mechanism including a first coupler 339 attached to the lid 326 and a second coupler 345 attached to a housing (z.e., the top wall 116 of the housing 102 of Figs. 1 and 2) or wall 324B of the incubation chamber 328. The first and second couplers 339, 345 may include male and female mating components, and may lock with a press-fit, interference fit, snap-fit, rotatable fit, adhesive, or other locking engagements. In some examples, the locking mechanism may include one or more of a hydraulic, spring, latch, grooves, ribs, a mainstay lock, clasp, a hook and loop, adhesive, or other male-female coupling mechanism that withstands an increase of pressure in the interior volume 333 of the incubation chamber 328. Additionally, and as shown in Fig. 8, the incubation chamber 328 includes a gasket 438 disposed at the opening (z.e., opening 118 in Figs. 1 and 2) of the incubation chamber 328. This gasket 438 provides an additional fluid-tight seal with the lid 326 when the lid 326 is secured in the closed position.

[0159] While the incubator system is configured for controlling environmental conditions of the incubator chamber for culturing an embryo have been described above to include control gas concentration (Figs. 1-4) or the internal pressure (Fig. 7) of the incubator chamber during different stages of embryonic development, in other examples, an incubator system may be configured to control both gas concentration (e.g., maintain 5% CO2 concentration and 5% O2 concentration) and pressure (e.g., in a range of approximately 1.0 atm to approximately 1.1 atm) during different embryonic stages of development. In other examples, an incubator system may be configured to control pressure and temperature during different embryonic stages of development. In other examples, an incubator system may be configured to control gas concentration, internal pressure, and / or internal temperature of the incubator chamber during different embryonic stages of development.

[0160] While the gas management systems 180, 380 of Figs. 4 and 8 have been described having recirculation lines 197A-B, 397, in other examples, a gas management system of an incubator may not have any recirculation lines, and instead the incubator chambers may vent to the surrounding environment.

[0161] While each of the gas mixing chambers 178A, 178B, 378 of the gas management systems 180, 380 of Figs. 4 and 8, respectively, has been described having a CO2 sensor 184 A, 184B, 384 and an O2 sensor 186 A, 186B, 386, in other examples, the incubator may include a CO2 sensor and an O2 sensor in (or otherwise coupled to) each incubation chamber,as shown in Figs. 9-11. While the control system 101 of the incubator of Figs. 1 and 6 has been described as being coupled to temperature sensors in the incubation chambers 128 and gas concentration sensors in the gas management system 180, in other examples, the control system 101 may additionally or alternatively be coupled to gas sensors coupled to the incubation chambers. For example, the control system 101 may be coupled to CO2 and O2 sensors 584, 586 coupled to each of the incubator chambers, as shown in Figs. 9 and 11.

[0162] Referring to Figs. 9-11, for example, the gas management system 580 is similar to the gas management system 180 described above and can be installed in the incubator 100 of Fig. 1. Elements of the gas management system 580 in Figs. 9-11 that are similar to the elements of the gas management system 180 of Fig. 4 are designated by the same reference numeral, incremented by 400. A description of many of these elements is abbreviated or even eliminated in the interest of brevity. The gas management system 580 differs from the gas management system 180 by controlling CO2 and O2 concentration in each incubation chamber 528 by controlling two separate inlet valves 537, 539, and by delivering pure gas directly into each incubation chamber 528.

[0163] Turning to Figs. 9-11, the incubator chamber 528 includes a first inlet 503, a second inlet 504, and exhaust out outlet 515, and an interior volume 533. The first and second inlets 503, 504 are connected to the sidewall portion 524A of the receptacle 520 for delivering N2 and CChto the interior volume 533 of the incubation chamber 528 through gas feed lines 594A, 594B. The first inlet 503 is operably coupled to a first valve 537, which is movable between an open position, in which a gas feed line 594A is in fluid communication with the interior volume 533, and a closed position, in which the gas feed line 594A is not in fluid communication with (or sealed from) the interior volume 533. The second inlet 504 is operably coupled to a valve 539, that is movable between an open position, in which the second gas feed line 594B is in fluid communication with the interior volume 533, and a closed position, in which the second gas feed line 595B is not in fluid communication with (or sealed from) the interior volume 533. The outlet 515 is found in an opposite sidewall 524B of the receptacle 520 for removing gas from the incubation chamber 528. As shown in Fig. 9, the CO2 source 574 is fluidly coupled to the first inlet valve 537, and the N2 source 576 is fluidly coupled to the second inlet valve 539. The outlet 515 is operably coupled to an outlet valve 543 and is movable between an open position, in which the interior volume 533 is in fluid communication with an environment, and a closed position, in which the incubation chamber 528 is sealed from the environment.

[0164] A monitoring system 198, such as the monitoring system 198 of Fig. 6, may be coupled to two gas sensors 584, 586 coupled to each incubation chamber 528 by hoses, as shown in Figs. 9 and 11. Further, a control system, such as the control system 101 of Fig. 6, may be operably coupled to one or more inlet and outlet valves 537, 539, 543 to control flow rate, pressure, and gas concentration in each incubation chamber 528.

[0165] While the incubation chamber 528 of Figs. 9-11 has been described having first and second inlet gas nozzles 503, 504, in other examples, an incubation chamber may include only one inlet gas nozzle.

[0166] In another example incubator system, an additional mixing chamber may be coupled to the gas management system 180 to provide a different gas concentration to the incubation chambers 128.

[0167] While the gas management systems of the previous examples have been described having separate N2 and CO2 sources, in other examples, a gas management system may instead include one or more premixed gas sources (e.g., premixed gas in cylinders) to provide the gas mixture to the incubator. For example, the premixed gas sources will include a mix of N2, CO2, and O2, each premixed gas source having a desired gas concentration composition for a particular embryonic growth stage.

[0168] While the gas management systems of the previous examples have been described as providing certain concentrations of CO2 to affect pH of media surrounding the biological sample in the incubator chamber, in other examples, a gas management system may instead be configured to control only O2 concentration to mirror the O2 concentration in the uterus and O2 concentration in the fallopian tubes (which has a higher O2 concentration than in the uterus). In this example, the CO2 concentration and / or internal pressure may be kept constant to minimize or avoid changes in pH. In this example, an O2 concentration may be in a range of approximately 4% (e.g., about 4.25% or more, about 4.5% or more, about 4.75% or more, about 5%) to approximately 6%(e.g., about 5.75% or less, about 5.5% or less, about 5.25% or less, about 5.0%) in the cleavage stage, and an O2 concentration may be in a range of approximately 1% (e.g., about 1.25% or more, about 1.5% or more, about 1.75% or more, about 2%) to approximately 4% (e.g., about 3.75% or less, about 3.5% or less, about 3.25% or less, about 3.0% or less, about 2.75% or less, about 2.5% or less, about 2.25% or less, about 2%) during the blastocyst stage.

[0169] This specification uses the term “configured” in connection with systems and computer program components. For a system of one or more computers to be configured to perform particular operations or actions means that the system has installed on it software,firmware, hardware, or a combination of them that in operation cause the system to perform the operations or actions. For one or more computer programs to be configured to perform particular operations or actions means that the one or more programs include instructions that, when executed by data processing apparatus, cause the apparatus to perform the operations or actions.

[0170] Embodiments of the subject matter and the functional operations described in this specification can be implemented in digital electronic circuitry, in tangibly-embodied computer software or firmware, in computer hardware, including the structures disclosed in this specification and their structural equivalents, or in combinations of one or more of them. Embodiments of the subject matter described in this specification can be implemented as one or more computer programs, i.e., one or more modules of computer program instructions encoded on a tangible non-transitory storage medium for execution by, or to control the operation of, data processing apparatus. The computer storage medium can be a machine- readable storage device, a machine-readable storage substrate, a random or serial access memory device, or a combination of one or more of them. Alternatively or in addition, the program instructions can be encoded on an artificially-generated propagated signal, e.g., a machine-generated electrical, optical, or electromagnetic signal that is generated to encode information for transmission to suitable receiver apparatus for execution by a data processing apparatus.

[0171] The term “data processing apparatus” refers to data processing hardware and encompasses all kinds of apparatus, devices, and machines for processing data, including by way of example a programmable processor, a computer, or multiple processors or computers. The apparatus can also be, or further include, special purpose logic circuitry, e.g., an FPGA (field programmable gate array) or an ASIC (application-specific integrated circuit). The apparatus can optionally include, in addition to hardware, code that creates an execution environment for computer programs, e.g., code that constitutes processor firmware, a protocol stack, a database management system, an operating system, or a combination of one or more of them.

[0172] A computer program, which may also be referred to or described as a program, software, a software application, an app, a module, a software module, a script, or code, can be written in any form of programming language, including compiled or interpreted languages, or declarative or procedural languages; and it can be deployed in any form, including as a stand-alone program or as a module, component, subroutine, or other unit suitable for use in a computing environment. A program may, but need not, correspond to afile in a file system. A program can be stored in a portion of a file that holds other programs or data, e.g., one or more scripts stored in a markup language document, in a single file dedicated to the program in question, or in multiple coordinated files, e.g., files that store one or more modules, sub-programs, or portions of code. A computer program can be deployed to be executed on one computer or on multiple computers that are located at one site or distributed across multiple sites and interconnected by a data communication network.

[0173] In this specification the term “engine” is used broadly to refer to a software-based system, subsystem, or process that is programmed to perform one or more specific functions. Generally, an engine will be implemented as one or more software modules or components, installed on one or more computers in one or more locations. In some cases, one or more computers will be dedicated to a particular engine; in other cases, multiple engines can be installed and running on the same computer or computers.

[0174] The processes and logic flows described in this specification can be performed by one or more programmable computers executing one or more computer programs to perform functions by operating on input data and generating output. The processes and logic flows can also be performed by special purpose logic circuitry, e.g., an FPGA or an ASIC, or by a combination of special purpose logic circuitry and one or more programmed computers.

[0175] Computers suitable for the execution of a computer program can be based on general or special purpose microprocessors or both, or any other kind of central processing unit. Generally, a central processing unit will receive instructions and data from a read-only memory or a random access memory or both. The essential elements of a computer are a central processing unit for performing or executing instructions and one or more memory devices for storing instructions and data. The central processing unit and the memory can be supplemented by, or incorporated in, special purpose logic circuitry. Generally, a computer will also include, or be operatively coupled to receive data from or transfer data to, or both, one or more mass storage devices for storing data, e.g., magnetic, magneto-optical disks, or optical disks. However, a computer need not have such devices. Moreover, a computer can be embedded in another device, e.g., a mobile telephone, a personal digital assistant (PDA), a mobile audio or video player, a game console, a Global Positioning System (GPS) receiver, or a portable storage device, e.g., a universal serial bus (USB) flash drive, to name just a few.

[0176] Computer-readable media suitable for storing computer program instructions and data include all forms of non-volatile memory, media and memory devices, including by way of example semiconductor memory devices, e.g., EPROM, EEPROM, and flash memorydevices; magnetic disks, e.g., internal hard disks or removable disks; magneto-optical disks; and CD-ROM and DVD-ROM disks.

[0177] To provide for interaction with a user, embodiments of the subject matter described in this specification can be implemented on a computer having a display device, e.g., a CRT (cathode ray tube) or LCD (liquid crystal display) monitor, for displaying information to the user and a keyboard and a pointing device, e.g., a mouse or a trackball, by which the user can provide input to the computer. Other kinds of devices can be used to provide for interaction with a user as well; for example, feedback provided to the user can be any form of sensory feedback, e.g., visual feedback, auditory feedback, or tactile feedback; and input from the user can be received in any form, including acoustic, speech, or tactile input. In addition, a computer can interact with a user by sending documents to and receiving documents from a device that is used by the user; for example, by sending web pages to a web browser on a user’s device in response to requests received from the web browser. Also, a computer can interact with a user by sending text messages or other forms of message to a personal device, e.g., a smartphone that is running a messaging application, and receiving responsive messages from the user in return.

[0178] Data processing apparatus for implementing machine learning models can also include, for example, special-purpose hardware accelerator units for processing common and compute-intensive parts of machine learning training or production, i.e., inference, workloads.

[0179] Machine learning models can be implemented and deployed using a machine learning framework, e.g., a TensorFlow framework, or a Jax framework.

[0180] Embodiments of the subject matter described in this specification can be implemented in a computing system that includes a back-end component, e.g., as a data server, or that includes a middleware component, e.g., an application server, or that includes a front-end component, e.g., a client computer having a graphical user interface, a web browser, or an app through which a user can interact with an implementation of the subject matter described in this specification, or any combination of one or more such back-end, middleware, or front-end components. The components of the system can be interconnected by any form or medium of digital data communication, e.g., a communication network. Examples of communication networks include a local area network (LAN) and a wide area network (WAN), e.g., the Internet.

[0181] The computing system can include clients and servers. A client and server are generally remote from each other and typically interact through a communication network.The relationship of client and server arises by virtue of computer programs running on the respective computers and having a client-server relationship to each other. In some embodiments, a server transmits data, e.g., an HTML page, to a user device, e.g., for purposes of displaying data to and receiving user input from a user interacting with the device, which acts as a client. Data generated at the user device, e.g., a result of the user interaction, can be received at the server from the device.

[0182] While the above discussed incubators have been described and illustrated with respect to certain dimensions, sizes, shapes, arrangements, materials, and methods, in some embodiments, an incubator that is otherwise substantially similar in construction and function to any of the above-discussed incubators may include one or more different dimensions, sizes, shapes, arrangements, configurations, and materials or may be utilized according to different methods.

[0183] While this specification contains many specific implementation details, these should not be construed as limitations on the scope of any disclosure or of what may be claimed, but rather as descriptions of features that may be specific to particular examples of particular disclosures. Certain features that are described in this specification in the context of separate examples can also be implemented in combination in a single example.Conversely, various features that are described in the context of a single example can also be implemented in multiple examples separately or in any suitable subcombination. Moreover, although features may be described herein as acting in certain combinations and even initially claimed as such, one or more features from a claimed combination can in some cases be excised from the combination, and the claimed combination may be directed to a subcombination or variation of a subcombination.

[0184] Similarly, while operations are depicted in the drawings in a particular order, this should not be understood as requiring that such operations be performed in the particular order shown or in sequential order, or that all illustrated operations be performed, to achieve desirable results. In certain circumstances, multitasking and parallel processing may be advantageous. Moreover, the separation of various system modules and components in the examples described herein should not be understood as requiring such separation in all examples, and it should be understood that the described program components and systems can generally be integrated together in a single product or packaged into multiple products.

[0185] Particular examples of the subject matter have been described. Other examples are within the scope of the following claims. For example, the actions recited in the claims can be performed in a different order and still achieve desirable results. As one example, theprocesses depicted in the accompanying figures do not necessarily require the particular order shown, or sequential order, to achieve desirable results. In certain implementations, multitasking and parallel processing may be advantageous.

Claims

CLAIMSWHAT IS CLAIMED IS:

1. A method of controlling an incubator, the method comprising: controlling an environmental condition of an incubation chamber for a first time period, wherein a pH value of media surrounding a biological specimen in a culture dish in the incubation chamber is affected by the environmental condition; and controlling the environmental condition of the incubation chamber for a second time period, wherein the pH value of the media surrounding a biological specimen in the culture dish during the second time period is different than the pH value of media during the first time period.

2. The method of claim 1, wherein controlling the environmental condition for the first time period comprises delivering a gas mixture having a CO2 concentration in a range of 4% to 7.0%.

3. The method of claim 1 or claim 2, wherein controlling the environmental condition for the first time period comprises delivering the gas mixture having a CO2 concentration in a range of 4.9% to 5.1%.

4. The method of any one of the preceding claims, wherein controlling the environmental condition for the second time period comprises delivering a gas mixture having a CO2 concentration in a range of 3% to 6%.

5. The method of any one of the preceding claims, wherein controlling the environmental condition for the second time period comprises delivering the gas mixture having a CO2 concentration in a range of 3.8% to 4.2%.

6. The method of any one of the preceding claims, wherein controlling the environmental condition for the first time period comprises delivering a gas mixture of N2 and CO2 for day one and day two of an embryonic growth cycle.

7. The method of claim 6, comprising premixing the gas mixture in a gas mixing chamber.

8. The method of any one of the preceding claims, wherein controlling the environmental condition for the second time period comprises delivering a different gas mixture of N2 and CO2 for day three, day four, and day five of the five to seven-day embryonic growth cycle.

9. The method of claim 8, comprising premixing the different gas mixture in a different gas mixing chamber.

10. The method of any one of the preceding claims, wherein controlling the environmental condition for the first time period comprises setting an internal pressure of the incubation chamber in a range of 111 kPa to 132 kPa.

11. The method of any one of the preceding claims, wherein controlling the environmental condition for the first time period comprises opening an inlet valve operably coupled to the incubation chamber, and closing an outlet valve operably coupled to the incubation chamber.

12. The method of any one of the preceding claims, wherein controlling the environmental condition for the second time period comprises setting an internal pressure of the incubation chamber in a range of 101 kPa to 111 kPa.

13. The method of claim 11 or claim 12, wherein controlling the environmental condition for the second time period comprises opening the outlet valve.

14. The method of any one of the preceding claims, wherein controlling the environmental condition for the first time period comprises increasing an internal pressure of the incubation chamber from about 101 kPa.

15. The method of claim 14, wherein controlling the environmental condition for the second time period comprises decreasing the internal pressure of the incubation chamber to a different predetermined pressure.

16. The method of claim 15, wherein controlling the environmental condition for the first time period comprises maintaining an internal pressure in a range of 110 kPa to 130 kPa, and controlling the environmental condition for the second time period comprises decreasing the internal pressure to 101 kPa.

17. The method of any one of the preceding claims, wherein controlling the environmental condition for the first time period comprises varying an internal pressure of the incubation chamber by delivering one or more of a high pressure N2 and a high pressure CO2.

18. The method of any one of the preceding claims, wherein controlling the environmental condition for the second time period comprises varying the internal pressure ofthe incubation chamber by delivering the one or more of a high pressure N2 and a high pressure CO2.

19. The method of any one of the preceding claims, comprising controlling an environmental condition of a second incubation chamber for a first time period, wherein a pH value of media surrounding a biological specimen in a culture dish in the second incubation chamber is affected by the environmental condition, wherein the biological specimen in the second incubation chamber is at a different growth stage than a growth stage of the biological specimen in the first incubation chamber; and controlling the environmental condition of the second incubation chamber for a second time period, wherein the pH value of media surrounding the biological specimen in the culture dish in the second incubation chamber during the second time period is different than the pH value of media in the second incubation chamber during the first time period.

20. A method of incubating one or more embryos, the method comprising: detecting a first gas concentration in a gas management system of an incubator using one or more of a Carbon dioxide gas (CO2) sensor and an O2 sensor; comparing a first detected gas concentration with a first stage O2 or CO2 concentration suitable for a first embryonic growth stage; delivering one or more of a Nitrogen gas (N2) and CO2 to an interior volume of an incubation chamber of the incubator to provide the first stage O2 or CO2 concentration in the interior volume of the incubation chamber; detecting a second gas concentration in the gas management system using the one or more of the CO2 sensor and the O2 sensor; comparing a second detected gas concentration with a second stage O2 or CO2 concentration suitable for a second embryonic growth stage; and delivering the one or more of the N2 and the CO2 to the interior volume of the incubation chamber to provide the second stage O2 or CO2 concentration in the interior volume of the incubation chamber.

21. The method of claim 20, wherein comparing the first detected gas concentration comprises comparing a first detected O2 concentration with an O2 concentration in a range of 4% to 6%.

22. The method of claim 20 or claim 21, wherein comparing the second detected gas concentration comprises comparing a second detected O2 concentration with an O2 concentration in a range of 4.5% to 5.5%.

23. The method of any one of claims 20 to 22, comprising detecting a first CO2 concentration in the gas management system using the CO2 sensor, and comparing a first detected CO2 concentration with a CO2 concentration in a range of 4% to 7%.

24. The method of claim 23, comprising detecting a second CO2 concentration in the gas management system using the CO2 sensor, and comparing a second detected CO2 concentration with a CO2 concentration in a range of 3% to 6%.

25. The method of any one of claims 20 to 24, wherein detecting the first gas concentration of the gas management system comprises detecting the first gas concentration of a first gas mixing chamber fluidly coupled to the incubation chamber.

26. The method of claim 25, wherein detecting the second gas concentration of the gas management system comprises detecting the second gas concentration of a second gas mixing chamber fluidly coupled to the incubation chamber.

27. The method of any one of claims 20 to 26, wherein detecting one or more of the first and second gas concentrations of the gas management system comprises detecting the one or more of the first and second gas concentrations of the interior volume of the incubation chamber.

28. The method of any one of claims 20 to 27, wherein delivering the one or more of the N2 and the CO2 to provide the first stage O2 or CO2 concentration comprises delivering the one or more of the N2 and the CO2 for days zero, one, and two of a five to seven-day embryonic growth cycle, wherein the first stage O2 or CO2 concentration comprises a CO2 concentration of about 5%.

29. The method of claim 28, wherein delivering the one or more of the N2 and the CO2 to provide the second stage O2 or CO2 concentration comprises delivering the one or more of the N2 and the CO2 for days three onward of the five to seven-day embryonic growth cycle, wherein the second stage O2 or CO2 concentration comprises a CO2 concentration of about 4%.

30. The method of claim 28 or claim 29, wherein delivering the one or more of the N2 and the CO2 to provide the first stage O2 or CO2 concentration comprises delivering a gas mixture of N2 and CO2 from a first gas mixing chamber.

31. The method of claim 30, wherein delivering the one or more of the N2 and the CO2 to provide the second stage O2 or CO2 concentration comprises delivering a different gas mixture from a second gas mixing chamber.

32. The method of any one of claims 20 to 31, comprising detecting a pressure value of the interior volume using a pressure sensor.

33. The method of claim 32, comprising comparing a detected pressure value with a first stage pressure value in a range of 1.1 atm to 1.3 atm, the first stage pressure value suitable for the first embryonic growth stage.

34. The method of claim 33, comprising modifying a pressure of the interior volume during days zero, one, and two of a five to seven-day embryonic growth cycle to provide the interior volume with the first stage pressure value.

35. The method of claim 34, comprising detecting a second pressure value of the interior volume using the pressure sensor, and comparing a second detected pressure value with a second stage pressure value in a range of 0.9 atm to 1.1 atm, the second stage pressure value suitable for the second embryonic growth stage.

36. The method of claim 35, comprising modifying the pressure of the interior volume during days three onward, of the five to seven-day embryonic growth cycle to provide the interior volume with the second stage pressure value.

37. The method of any one of claims 34 to 36, wherein modifying the pressure comprises varying the pressure of the incubation chamber.

38. The method of any one of claims 34 to 37, wherein modifying the pressure comprises delivering one or more of a high pressure N2 and a high pressure CChto the interior volume of the incubation chamber.

39. The method of claim 38, wherein delivering one or more of the high pressure N2 and the high pressure CO2 comprises delivering one or more of the high pressure N2 and the high pressure CO2 at more than 120 kPa.

40. The method of any one of claims 34 to 39, wherein modifying the pressure comprises increasing the pressure of the incubation chamber from 1 atm to 1.2 atm during the first embryonic growth stage.

41. The method of any one of claims 20 to 40, comprising recycling a gas mixture delivered to the incubation chamber to a mixing chamber.

42. The method of claim 41, comprising recycling a different gas mixture delivered to the incubation chamber to a second gas mixing chamber.

43. The method of any one of claims 20 to 42, wherein the steps of detecting, comparing, and delivering are performed with respect to a second incubation chamber of the incubator.

44. An incubator system for assisted reproductive technologies, the incubator system comprising: a nitrogen gas (N2) source; a carbon dioxide gas (CO2) source; a gas feed line fluidly coupleable with one or more of the N2 source and the CO2 source; an incubation chamber operably coupleable to the gas feed line, the incubation chamber comprising an inlet, an outlet, and an interior volume disposed between the inlet and the outlet, the interior volume being sized to receive a dish holding a biological specimen; an inlet valve operably coupleable to the inlet of the incubation chamber, the inlet valve movable to an open position in which the gas feed line is in fluid communication with the interior volume; and an outlet valve operably coupleable to the outlet of the incubation chamber, the outlet valve movable to an open position to remove a gas mixture from the interior volume; and wherein a pH value of media surrounding a biological specimen disposed in the interior volume of the incubation chamber is controllable by operating one or more of the inlet valve and the outlet valve.

45. The incubator system of claim 44, comprising a processor configured to control an environmental condition of the incubation chamber for a first time period, wherein a pH value of media surrounding a biological specimen in a culture dish in the incubation chamber is affected by the environmental condition; andwherein the processor is configured to control the environmental condition of the incubation chamber for a second time period, wherein the pH value of media during the second time period is different than the pH value of media during the first time period.

46. The incubator system of claim 44, comprising a gas mixing chamber fluidly coupleable to the N2 source and the CO2 source, the gas feed line fluidly coupleable to the gas mixing chamber.

47. The incubator system of claim 46, comprising a second gas feed line and a second gas mixing chamber fluidly coupleable to the N2 source and CO2 source and to the second gas feed line.

48. The incubator system of claim 47, wherein the inlet valve is movable to a second open position in which the second gas feed line is in fluid communication with the interior volume of the incubation chamber, and the gas feed line is fluidly isolated from the mixing chamber.

49. The incubator system of claim 47 or claim 48, wherein when the outlet valve is in the open position, the interior volume is in fluid communication with the gas mixing chamber.

50. The incubator system of any one of claims 47 to 49, wherein the outlet valve is movable to a second open position in which the interior volume is in fluid communication with the second gas mixing chamber.

51. The incubator system of any one of claims 47 to 50, wherein the gas mixing chamber has a CO2 gas concentration in a range of 4% CO2 to 7% CO2 and an O2 concentration in a range of 4% O2 to 6% O2.

52. The incubator system of any one of claims 47 to 51, wherein the second gas mixing chamber has a CO2 gas concentration in a range of 3% CO2 to 6% CO2 and an O2 concentration in a range of 4% O2 to 6% O2.

53. The incubator system of any one of claims 47 to 52, comprising one or more of a CO2 sensor and an O2 sensor coupled to the gas mixing chamber.

54. The incubator system of any one of claims 47 to 53, comprising one or more of a CO2 sensor and an O2 sensor coupled to the second gas mixing chamber.

55. The incubator system of any one of claims 44 to 54, comprising one or more of a CO2 sensor and an O2 sensor coupled to the interior volume of the incubation chamber.

56. The incubator system of any one of claims 44 to 55, comprising a pressure sensor coupled to the interior volume of the incubation chamber.

57. The incubator system of any one of claims 44 to 56, wherein the incubation chamber comprises a lid and a locking mechanism coupled to the lid, the locking mechanism configured to seal the interior volume from atmosphere in a locked configuration.

58. The incubator system of any one of claims 44 to 57, wherein the incubation chamber comprises a second inlet operably coupled to a second inlet valve.

59. The incubator system of claim 58, comprising a second gas feed line operably coupled to the second inlet valve and in fluid communication with the N2 source, wherein the gas feed line is in fluid communication with the CO2 source and operably coupled to the inlet valve.

60. The incubator system of any one of claims 44 to 59, comprising a plurality of incubation chambers, wherein an environmental condition of each of the plurality of incubation chambers is independently controllable.

61. An incubator system for assisted reproductive technologies, the incubator system comprising: a Nitrogen gas (N2) source; a Carbon dioxide gas (CO2) source; a gas mixing chamber fluidly coupled to the N2 source and the CO2 source, the gas mixing chamber including an inlet and an outlet; a gas feed line coupled to the outlet of the gas mixing chamber; a gas recirculation line coupled to the inlet of the gas mixing chamber; an incubation chamber fluidly coupled to the gas mixing chamber, the incubation chamber comprising an inlet, an outlet, and an interior volume disposed between the inlet and the outlet, the interior volume being sized to receive a dish holding a biological specimen; an inlet valve operably coupled to the inlet of the incubation chamber, the inlet valve movable to an open position in which the gas feed line is in fluid communication with the interior volume of the incubation chamber, and to a different position in which the gas feed line is not in fluid communication with the interior volume; and wherein a pH value of media surrounding a biological specimen disposed in the interior volume of the incubation chamber is controllable by operating the inlet valve.

62. The incubator system of claim 61, comprising a processor configured to control an environmental condition of the incubation chamber for a first time period, wherein a pH value of media surrounding a biological specimen in a culture dish in the incubation chamber is affected by the environmental condition; and wherein the processor is configured to control the environmental condition of the incubation chamber for a second time period, wherein the pH value of media during the second time period is different than the pH value of media during the first time period.

63. The incubator system of claim 61, comprising a second gas feed line and a second gas mixing chamber fluidly coupleable to the N2 source and CO2 source and to the second gas feed line.

64. The incubator system of claim 63, wherein the inlet valve is movable to a second open position in which the second gas feed line is in fluid communication with the interior volume of the incubation chamber, and the gas feed line is fluidly isolated from the mixing chamber.

65. The incubator system of claim 63 or claim 64, comprising an outlet valve operably coupled to the outlet of the incubation chamber, the outlet valve movable to an open position in which the gas recirculation line is in fluid communication with the interior volume, and a different position in which the gas recirculation line is not in fluid communication with the interior volume; and wherein the outlet valve is movable to a second open position,- in which the interior volume is in fluid communication with the second gas mixing chamber.

66. The incubator system of any one of claims 63 to 65, wherein the gas mixing chamber has a CO2 gas concentration in a range of 4% CO2 to 7% CO2 and an O2 concentration in a range of 4% O2 to 6% O2.

67. The incubator system of claim 66, wherein the second gas mixing chamber has a CO2 gas concentration in a range of 3% CO2 to 6% CO2 and an O2 concentration in a range of 4% O2to 6% O2.

68. The incubator system of any one of claims 61 to 67, comprising one or more of a CO2 sensor and an O2 sensor coupled to the gas mixing chamber.

69. The incubator system of any one of claims 63 to 68, comprising one or more of a CO2 sensor and an O2 sensor coupled to the second gas mixing chamber.

70. The incubator system of any one of claims 61 to 69, comprising a pressure sensor coupled to the interior volume of the incubation chamber.

71. The incubator system of any one of claims 61 to 70, wherein the incubation chamber comprises a lid and a locking mechanism coupled to the lid, the locking mechanism configured to seal the interior volume from atmosphere in a locked configuration.

72. The incubator system of any one of claims 61 to 71, comprising a plurality of incubation chambers, wherein an environmental condition of each of the plurality of incubation chambers is independently controllable.

73. An incubator system for assisted reproductive technologies, the incubator comprising: a Nitrogen gas (N2) source; a Carbon dioxide gas (CO2) source; a first gas mixing chamber fluidly coupled to the N2 source and the CO2 source and configured for containing a first gas mix having a CO2 gas concentration in a range of 4% CO2 to 7% CO2 and an oxygen gas (O2) concentration in a range of 4% O2 to 6% O2; a second gas mixing chamber fluidly coupled to the N2 source and the CO2 source and configured for containing a second gas mix having a CO2 gas concentration in a range of 4% CO2 to 7% CO2 and an O2 concentration in a range of 1% O2 to 4% O2; an incubation chamber fluidly coupled to the first gas mixing chamber and the second gas mixing chamber, the incubation chamber comprising an inlet, an outlet, and an interior volume disposed between the inlet and the outlet, the interior volume configured for receiving a dish holding a biological specimen; a first gas feed line coupling the first gas mixing chamber and the inlet; a second gas feed line coupling the second gas mixing chamber and the inlet; and wherein an O2 concentration of the interior volume of the incubation chamber is controllable by delivering one or more of the first gas mix and the second gas mix through the inlet of the incubation chamber.

74. The incubator system of claim 73, comprising one or more of a CO2 sensor and an O2 sensor coupled to the first gas mixing chamber, and one or more of a CO2 sensor and an O2 sensor coupled to the second gas mixing chamber.

75. The incubator system of claim 73 or claim 74, comprising a gas recirculation line coupling the outlet of the incubation chamber to the first gas mixing chamber.

76. The incubator system of claim 75, comprising a second gas recirculation line coupling the outlet of the incubation chamber to the second gas mixing chamber.

77. The incubator system of any one of claims 73 to 76, comprising an inlet valve movable to a first open position in which the interior volume is in fluid communication with the first gas mixing chamber and isolated from the second gas mixing chamber, and a second open position in which the interior volume is in fluid communication with the second gas mixing chamber and isolated from the first gas mixing chamber.

78. The incubator system of any one of claims 73 to 77, comprising an outlet valve movable to a first open position in which the interior volume is in fluid communication with the first gas mixing chamber, and a second open position in which the interior volume is in fluid communication with the second gas mixing chamber.

79. The incubator system of any one of claims 73 to 78, comprising a plurality of incubation chambers, wherein a O2 concentration of each of the plurality of incubation chambers is separably controllable.

80. The incubator system of any one of claims 73 to 79, comprising a plurality of inlet valves and a plurality of incubation chambers, each inlet valve operably coupled to an inlet of a different incubation chamber, wherein each of the plurality of inlet valves is separably controllable.

81. The incubator system of any one of claims 73 to 80, comprising a processor configured to control an environmental condition of the incubation chamber for a first time period, and to control the environmental condition of the incubation chamber for a second time period.

82. An incubator system for assisted reproductive technologies, the incubator system comprising: a first premixed gas source comprising Oxygen gas (O2), Nitrogen gas (N2), and Carbon dioxide gas (CO2); a second premixed gas source comprising O2, N2, and CO2; a first gas feed line coupled to the first premixed gas source; a second gas feed line coupled to the second premixed gas source; a gas recirculation line coupled to the first premixed gas source;an incubation chamber fluidly coupled to the first premixed gas source, the incubation chamber comprising an inlet, an outlet, and an interior volume disposed between the inlet and the outlet, the interior volume being sized to receive a dish holding a biological specimen; an inlet valve operably coupled to the inlet of the incubation chamber, the inlet valve movable to an open position in which the first gas feed line is in fluid communication with the first premixed gas source, and to a different position in which the first gas feed line is not in fluid communication with the first premixed gas source; wherein a pH value of media surrounding a biological specimen disposed in the interior volume of the incubation chamber is controllable by operating the inlet valve; and wherein the inlet valve is movable to a second open position in which the second gas feed line is in fluid communication with the interior volume of the incubation chamber, and the first gas feed line is fluidly isolated from the first premixed gas source.

83. The incubator system of claim 82, comprising a a processor configured to control an environmental condition of the incubation chamber for a first time period, wherein a pH value of media surrounding a biological specimen in a culture dish in the incubation chamber is affected by the environmental condition; and wherein the processor is configured to control the environmental condition of the incubation chamber for a second time period, wherein the pH value of media during the second time period is different than the pH value of media during the first time period.

84. The incubator system of claim 82, comprising an outlet valve operably coupled to the outlet of the incubation chamber, the outlet valve movable to an open position in which the gas recirculation line is in fluid communication with the interior volume, and a different position in which the gas recirculation line is not in fluid communication with the interior volume.

85. The incubator system of claim 84, wherein the outlet valve is movable to a second open position in which the interior volume is in fluid communication with the second premixed gas source.

86. The incubator system of claim 84 or 85, wherein the first premixed gas source has a CO2 gas concentration in a range of 4% CO2 to 7% CO2 and an O2 concentration in a range of 4% O2 to 6% O2.

87. The incubator system of claim 86, wherein the second premixed gas source has a CO2 gas concentration in a range of 3% CO2 to 6% CO2 and an O2 concentration in a range of 4% O2 to 6% O2.

88. The incubator system of any one of claims 82 to 87, comprising one or more of a CO2 sensor and an O2 sensor coupled to the first premixed gas source.

89. The incubator system of any one of claims 84 to 88, comprising one or more of a CO2 sensor and an O2 sensor coupled to the second premixed gas source.

90. The incubator system of any one of claims 82 to 89, comprising a pressure sensor coupled to the interior volume of the incubation chamber.

91. The incubator system of any one of claims 82 to 90, wherein the incubation chamber comprises a lid and a locking mechanism coupled to the lid, the locking mechanism configured to seal the interior volume from atmosphere in a locked configuration.

92. The incubator system of any one of claims 82 to 91, comprising a plurality of incubation chambers, wherein an environmental condition of each of the plurality of incubation chambers is independently controllable.

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