Patent Description:
Low temperature specimen carriers, such as cryopreservation devices, are used in the field of assisted reproductive technology (ART) to store and preserve living reproductive specimens (e.g., oocytes, embryos, and blastocysts). Cryopreservation refers to a process in which specimens are preserved over extended periods of time by cooling to sub-zero temperatures. For example, a cryopreservation device can house and support specimens undergoing vitrification, which is the rapid transition of a substance from a liquid phase to a solid phase (e.g., glass) without the formation of ice crystals within cells of the specimen.

Typical protocols for vitrifying a reproductive specimen include accessing a carrier (e.g., a petri dish, a test tube, or flask) in which the specimen is disposed multiple times to expose the specimen to multiple processing solutions. Such protocols further include subsequently transferring the specimen to a cryopreservation device, and then exposing the cryopreservation device, containing the specimen therein, to a cooling medium (e.g., liquid nitrogen) to cause the cells of the specimen to rapidly cool to a glass state before ice crystals can form within the cells. The cryopreservation device can be stored in the cooling medium until the specimen is ready to be used in reproductive procedures.

Examples of known protocols are described in the following references:.

The above references relate to vitrification of embryos and disclose specimen containers preloaded with solutions that are initially separated by air.

In general, this disclosure relates to specimen containers configured for preparation and storage of a specimen in a low temperature substance and relates to associated methods. Such specimen containers can be used for preserving living specimens in a viable state over a prolonged period of time.

Claim <NUM> provides a specimen container according to the present invention. In the following, the expression "first specimen processing medium" is used synonymously with "equilibration solution", and "second specimen processing medium" with "vitrification solution".

Embodiments may include one or more of the following features.

In some embodiments, the elongate member is a capillary tube.

In some embodiments, the first and second specimen processing mediums are spaced apart from each other by the barrier within the lumen of the elongate member in a preloaded configuration of the specimen container.

In certain embodiments, the elongate member includes a first portion having constant width and a second portion having a variable width.

In certain embodiments, the second portion is a tapered portion.

In some embodiments, the second specimen processing medium is denser than the first specimen processing medium.

In certain embodiments, the first specimen processing medium has a volume of about <NUM>µL to about <NUM>µL.

In some embodiments, the second specimen processing medium has a volume of about <NUM>µL to about <NUM>µL.

In certain embodiments, the specimen container further includes one or more additional specimen processing mediums.

In some embodiments, the barrier is a fluid.

In certain embodiments, the fluid includes air.

In some embodiments, the barrier includes a valve.

In certain embodiments, the barrier is an inert solid that undergoes a solid to liquid phase change at a temperature of about <NUM>.

In some embodiments, the barrier, in a liquid phase, is less dense than the first and second specimen processing mediums.

In certain embodiments, the barrier includes a clamping mechanism disposed external to the elongate member.

In some embodiments, the elongate member includes a flexible tube.

In some embodiments, a diameter of the elongate member varies in a stepwise manner along an axis of the elongate member.

In certain embodiments, the specimen container further includes an electronic identification label.

In some embodiments, a proximal end of the elongate tube and is wider than a central portion of the elongate tube.

In certain embodiments, the elongate tube defines a sidewall opening located proximal to the first processing medium.

In certain embodiments, the specimen container further includes a plug configured to fit within the lumen of the elongate tube and a specimen carrier that extends from the plug.

In some embodiments, one or both of the first and second processing mediums includes magnetic nanoparticles.

In certain embodiments, the specimen container is formed of a material that can mechanically withstand a temperature of about -<NUM> or less for at least about <NUM> years.

Claim <NUM> provides a cryogenic device according to the present invention.

Claim <NUM> provides a vitrification system according to the present invention.

Claim <NUM> provides a method of cryogenically processing a specimen according to the present invention.

In some embodiments, the method further includes passing the specimen through a proximal opening in the specimen container.

In certain embodiments, the proximal opening is located at an end of the specimen container.

In some embodiments, the proximal opening is located along a sidewall of the specimen container.

In certain embodiments, the method further includes sealing the proximal opening of the specimen container after depositing the specimen within the lumen of the specimen container.

In some embodiments, the method further includes merging the first and second processing mediums to form a combined processing medium within the lumen of the elongate member.

In certain embodiments, the predetermined period of time is a first predetermined period of time, and the method further includes exposing the specimen to the combined processing medium for a second predetermined period of time.

In some embodiments, the method further includes spinning the specimen container about an axis of the specimen container while the specimen is contained within the specimen container.

In certain embodiments, the method further includes revolving the specimen container around a revolution axis while the specimen is contained within the specimen container.

In some embodiments, the first processing medium includes magnetic nanoparticles.

In certain embodiments, the method further includes applying a magnetic force to the magnetic nanoparticles.

In some embodiments, the method further includes immersing the specimen container within liquid nitrogen.

In certain embodiments, the method further includes exposing the specimen to a temperature of about -<NUM> or less while the specimen is disposed within the specimen container.

In some embodiments, the method further includes vitrifying the specimen within the specimen container.

In certain embodiments, the method further includes thawing the specimen within the specimen container.

In some embodiments, the method further includes dispelling the specimen from the specimen container.

In certain embodiments, the method further includes reading an electronic identification label of the specimen container.

In some embodiments, the specimen includes one or more reproductive cells.

Embodiments may provide one or more of the following advantages.

The specimen container is designed to exploit mass properties of a specimen with respect to mass properties of various processing media. Accordingly, the lumen of the specimen container is internally preloaded with multiple fluids to which the specimen will be exposed during a cryopreparation process. In particular, the specimen container can be preloaded with an equilibration solution of relatively low density and a vitrification solution of relatively high density that are separated by a separation fluid <NUM>. Such separation of the equilibration solution and the vitrification solution enables appropriate processing of the specimen (e.g., sequential exposure of the specimen to particular solutions for desired periods of time) during vitrification protocols.

Furthermore, owing to a preloaded state of the equilibration solution and the vitrification solution within the specimen container, a specimen can be prepared for vitrification within a single, isolated environment (e.g., the lumen of the specimen container) without being exposed to contamination, mechanical damage (e.g., from a micropipette or other specimen holding or fluid delivery device), or other accidental mishandling that may otherwise occur when a container that houses a specimen is accessed multiple times to deliver and remove various processing mediums or when a specimen is moved to various containers during an ART process. In this regard, the specimen containers discussed herein are easy-to-use devices that enable a user to simply deposit a specimen within a container and then place the container within a system console or a centrifuge to carry out certain stages of an ART protocol. Accordingly, the user can avoid steps involving adding and removing multiple different fluids to a specimen container. Additionally, the specimen container has as geometry that optimizes storage density, such that the specimen container occupies little space. A construction of the specimen container also has a low thermal capacity, such that the specimen container experiences raid cooling and warming rates, which promotes wellness of tissue specimens contained therein.

<FIG> illustrates a specimen container <NUM> that can be used to prepare a specimen according to a biological or other protocol and to subsequently store the specimen in a low temperature substance. In particular, the specimen container <NUM> is a cryogenic device that is configured for cryopreparation and cryopreservation of a specimen in a viable and vitrified state within the low temperature substance until the specimen is desired for use (e.g., over a period of up to about <NUM> years). The specimen may be a single cell, a collection of free (e.g., unattached) cells, or a collection of attached cells (e.g., a multicellular tissue).

Example specimens include reproductive specimens (e.g., oocytes, zygotes, embryos, blastocysts, and gastrulae) and other, non-reproductive specimens (e.g., T-cells and blood cells). The specimen may be a mammalian sample or a non-mammalian sample. In some examples, the specimen is an agricultural specimen, such as canola. In some instances, the specimen is a non-biological specimen, such as various chemicals or other non-biological specimens. The low temperature substance (e.g., liquid nitrogen, cryogenic plasma, or liquid helium) typically has a temperature of about -<NUM> to about - <NUM> (e.g., about -<NUM> for liquid nitrogen) and maintains the specimen in a vitrified state.

Referring to <FIG> and <FIG>, the specimen container <NUM> includes an elongate tube <NUM>, a proximal closure <NUM> that hermetically seals a proximal end <NUM> of the elongate tube <NUM>, and a distal closure <NUM> that hermetically seals a distal end <NUM> of the elongate tube <NUM>. The elongate tube <NUM> is a thin capillary tube of very small diameter (e.g., having an internal diameter on the order of <NUM>-<NUM> m). The elongate tube <NUM> has a substantially constant diameter along a main portion <NUM> (e.g., a cylindrical portion) and has a variable diameter that gradually decreases along a tapered portion <NUM> that extends from the main portion <NUM> to the distal end <NUM>.

The proximal closure <NUM> is a cap that is designed to surround the proximal end <NUM> of the elongate tube <NUM>. The proximal closure <NUM> can be reversibly installed and removed from the proximal end <NUM> to seal the proximal end <NUM> and to open the proximal end <NUM> to allow proximal access to the elongate tube <NUM>, respectively. The distal closure <NUM> is a single-use seal (e.g., a melt seal, a fold, glue or adhesive, or an occluding member) that can be removed (e.g., cut or otherwise separated) from the distal end <NUM> of the elongate tube <NUM> to allow material to pass distally out of the elongate tube <NUM>.

The main portion <NUM> of the elongate tube <NUM> typically has a length of about <NUM> to about <NUM> (e.g., about <NUM>), an outer diameter of about <NUM> to about <NUM> (e.g., about <NUM>), and a wall thickness of about <NUM> to about <NUM> (e.g., about <NUM>). The tapered portion <NUM> of the elongate tube <NUM> typically has a length of about <NUM> to about <NUM> (e.g., about <NUM>), a maximum outer diameter that is adjacent and equal to the outer diameter of the main portion <NUM>, a minimum outer diameter (e.g., at the distal end <NUM> of the elongate tube <NUM>) of about <NUM> to about <NUM> (e.g., about <NUM>), and a wall thickness of about <NUM> to about <NUM> (e.g., about <NUM>). A lumen of the elongate tube <NUM>, at a smallest inner diameter, is large enough to accommodate a specimen, which typically has a diameter or a width in a range of about <NUM> to about <NUM>. A geometry and a construction (e.g., a thin and small profile) of the elongate tube <NUM> are configured to increase (e.g., maximize) heat transfer and to reduce (e.g., minimize) thermal mass to provide suitable cooling and warming rates of the specimen container <NUM> during ART protocols. The specimen container <NUM> typically has a total length (e.g., including lengths of the elongate tube <NUM>, the proximal closure <NUM>, and the distal closure <NUM>) of about <NUM> to about <NUM> (e.g., about <NUM>).

The elongate tube <NUM> may be manufactured via an injection molding process, a casting process, or an extrusion process. The elongate tube <NUM> is typically made of one or more materials that can withstand the low temperature substance, including but not limited to polymers such as polystyrene, polypropylene, polyvinyl acetate, and polycarbonate, and fluoropolymers. The elongate tube <NUM> is also typically transparent or translucent to allow viewing of the specimen through the wall of the elongate tube <NUM>. The proximal and distal closures <NUM>, <NUM> may be manufactured via radio frequency (RF) or ultrasonic sealing and are typically made of one or more materials that can withstand the low temperature substance, including but not limited to polymers such as polystyrene, polypropylene, polyvinyl acetate, and polycarbonate, and fluoropolymers.

The specimen container <NUM> is designed to exploit mass properties (e.g., density or fluid mechanics) of a specimen with respect to mass properties of various processing media. Accordingly, the lumen of the elongate tube <NUM> is internally preloaded with multiple fluids to which the specimen will be exposed during a cryopreparation process. In some implementations, for example, the elongate tube <NUM> is preloaded with an equilibration solution <NUM> (e.g., a cryoprotectant of relatively low density) and a vitrification solution <NUM> (e.g., a cryoprotectant of relatively high density) that are separated by a separation fluid <NUM> (e.g., an air bubble or an immiscible media). Such separation of the equilibration solution <NUM> and the vitrification solution <NUM> enables appropriate processing of the specimen (e.g., sequential exposure of the specimen to particular solutions for desired periods of time) during vitrification protocols. The elongate tube <NUM> is further preloaded with a proximal air pocket <NUM> that separates the equilibration solution <NUM> from the proximal closure <NUM> and a distal air pocket <NUM> (e.g., occupying a portion of an interior volume of the tapered portion <NUM> of the elongate tube <NUM>) that separates the vitrification solution <NUM> from the distal closure <NUM>.

Example equilibration solutions <NUM> include non-essential and essential amino acids, gentamicin sulfate (<NUM>/L), <NUM>% (v/v) each of DMSO and ethylene glycol and <NUM>/mL human albumin Such equilibration solutions <NUM> typically have a density in a range of about <NUM>/mL to about <NUM>/mL. The volume of the equilibration solution <NUM> within the elongate tube <NUM> is typically about <NUM>µL to about <NUM>µL. Example vitrification solutions <NUM> include non-essential amino acids, gentamicin sulfate (<NUM>/L), <NUM>% (v/v) each of DMSO and ethylene glycol, <NUM>/mL human albumin, and <NUM> sucrose. Such vitrification solutions <NUM> typically have a density in a range of about <NUM>/mL to about <NUM>/mL, such that the vitrification solution <NUM> is typically more dense than the equilibration solution <NUM>. The volume of the vitrification solution <NUM> within the elongate tube <NUM> is typically about <NUM>µL to about <NUM>µL. The volume of the separation fluid <NUM> is typically about <NUM>µL to about <NUM>µL. The separation fluid <NUM> typically has a density that is less than densities of the equilibration solution <NUM> and the vitrification solution <NUM>. For example, the separation fluid <NUM> typically has a density in a range of about <NUM>/mL to about <NUM>×<NUM>-<NUM> g/mL. The equilibration solution <NUM> and the vitrification solution <NUM> are typically axially spaced about <NUM> to about <NUM> apart from each other within the elongate tube <NUM> (e.g., according to the volume of the separation fluid <NUM> and an inner diameter of the elongate tube <NUM>). The specimen to be placed in the container typically has a density that is different from densities of the equilibration solution <NUM> and the vitrification solution <NUM>. The specimen typically has a density that is slightly greater than about <NUM>/mL outside of the solutions <NUM>, <NUM>, but that rapidly changes upon exposure to the solutions <NUM>, <NUM>. For example, initially, the specimen nearly floats in the equilibration solution <NUM>, but becomes denser as cells of the specimen are hydrated by the equilibration solution <NUM>.

Referring to <FIG>, the specimen container <NUM> further includes an identification (ID) label <NUM> attached to the elongate tube <NUM> near the proximal end <NUM>. The ID label <NUM> can be a radio-frequency identification (RFID) tag (e.g., including an internal antenna) that includes machine readable information. Additionally, human readable information may be written on an outer surface of the ID label <NUM>. Either or both of the machine readable information and the human readable information may include various patient data, such as a name, a birthdate, and a unique reference code (e.g., an alphanumeric sequence). The ID label <NUM> of the specimen container <NUM> can be detected and read by a scanning component of a system console, as will be discussed in more detail below with respect to <FIG>.

The specimen container <NUM> is a sterile, single-use device that is non-toxic to specimens contained therein. The specimen container <NUM> is typically packaged as a single unit, and both the specimen container <NUM> and the packaging will remain sterile for a guaranteed shelf-life of the specimen container <NUM>. The total length of the specimen container <NUM> typically allows the specimen container <NUM> to fit within standard storage containers and other standard equipment used in ART protocols.

In some embodiments, a system console including various ART components can be used to process a specimen contained within the specimen container <NUM>. For example, referring to <FIG>, a system console <NUM> includes a base housing <NUM>, a receptacle <NUM> that can spin within an interior pocket <NUM> of the base housing <NUM>, a reader component <NUM> (e.g., an RFID antenna or another type of reader component, illustrated schematically) that is programmed to read the ID label <NUM> of the specimen container <NUM>, and a cooler <NUM> that is slidable within a drawer <NUM> of the base housing <NUM> and that is configured to contain a low temperature substance. The system console <NUM> further includes a lid <NUM> that is openable to allow access to the receptacle <NUM>, a user interface screen <NUM> positioned along a front side of the lid <NUM>, a timer <NUM> (illustrated schematically), and a control module <NUM> (illustrated schematically) that is programmed to control various features and functionalities of the system console <NUM>. The reader component <NUM>, the timer <NUM>, and the control module <NUM> may be positioned at respective locations within the system console <NUM> that are suitable for their respective functions. In some embodiments, the system console <NUM> further includes an accessory tube <NUM> that is sized to surround the specimen container <NUM> and to be received in the receptacle <NUM>.

The base housing <NUM> is configured to support the receptacle <NUM> and the lid <NUM>, to receive the cooler <NUM>, and to rest atop a floor or another flat surface. The receptacle <NUM> is provided as an elongate channel that is sized to receive the specimen container <NUM> at an entry opening <NUM>. The reader component <NUM> can detect a presence of the specimen container <NUM> within the receptacle <NUM> by reading the ID label <NUM> (e.g., the RFID tag) and can communicate such detection to the control module <NUM>, which can cause the timer <NUM> to be activated. According to one or more signals received from the control module <NUM>, the receptacle <NUM> can spin within the interior pocket <NUM> about a spin axis <NUM> of the receptacle <NUM>. Such spinning causes the specimen to move within the specimen container <NUM> along a radial axis <NUM> of the receptacle <NUM> toward the distal end <NUM> of the elongate tube <NUM>. According to one or more signals received from the control module <NUM>, an adjustable exit opening <NUM> of the receptacle <NUM> can be closed or constricted to support the specimen container <NUM> during spinning. The exit opening <NUM> can also be opened or enlarged to release the specimen container <NUM> downward through an exit channel <NUM> of the base housing <NUM> and into the cooler <NUM> following spinning for vitrification and storage of the specimen container <NUM> within the low temperature substance contained within the cooler <NUM>.

One or more storage containers may be disposed within the cooler <NUM> for receiving the specimen container <NUM>. In some embodiments, the cooler <NUM> can be slid in and out of the drawer <NUM> of the base housing <NUM> in an automated manner according to one or more signals received from the control module <NUM> to allow a user to check a level of the low temperature substance (e.g., which can be susceptible to evaporation) and/or to refill the cooler <NUM> with the low temperature substance. In some embodiments, the cooler <NUM> is configured to be manually slid in and out of the drawer <NUM> via a handle <NUM>. In some embodiments, the system console <NUM> includes a sensor that can detect the level of the low temperature substance within the cooler <NUM>.

The lid <NUM> is manually movable (e.g., pivotable, slidable, or removable) with respect to the base housing <NUM> to allow access to the receptacle <NUM>. The user interface screen <NUM> allows a user to input several parameters that govern operation of the system console <NUM> to vitrify the specimen <NUM>, such as a stage of the specimen <NUM> (e.g., an oocyte or a blastocyst protocol selection). The user interface screen <NUM> may be an integrated touchscreen or a touchless screen associated with tactile control elements, such as buttons, knobs, dials, or the like. The control module <NUM> includes one or more processors that are in communication with and/or are programmed to control various actuators and sensors of the system console <NUM> related to various automated features, such as receiving and instantiating user selections input at the user interface screen <NUM>, reading the ID label <NUM> of the specimen container <NUM>, executing the timer <NUM>, spinning the receptacle <NUM> at a specified spin speed for a specified duration, adjusting the exit opening <NUM> of the receptacle <NUM>, sliding the cooler <NUM> along the drawer <NUM>, detecting the level of the low temperature substance, detecting an open/closed state of the lid <NUM>, and providing audible and/or visual feedback regarding a progression of the process.

In some embodiments, the base housing <NUM> and the lid <NUM> of the system console <NUM> have a length of about <NUM> to about <NUM> and a width of about <NUM> to about <NUM>. In some embodiments, the base housing <NUM> has a height of about <NUM> to about <NUM>, and the lid <NUM> has a height of about <NUM> to about <NUM>, such that the system console <NUM> has a total height (e.g., when the lid <NUM> is closed) of about <NUM> to about <NUM>. In some embodiments, the system console <NUM> (e.g., absent the low temperature substance) has a weight in a range of about <NUM> to about <NUM> and is typically stored on a laboratory floor, a storage facility floor, a table, or a countertop, that has an ambient environmental temperature of about <NUM> to about <NUM>. In some embodiments, the receptacle <NUM> has a length of about <NUM> to about <NUM>. In some embodiments, the receptacle <NUM> is sized to hold one specimen container <NUM>, such that the receptacle <NUM> has an internal diameter of about <NUM> to about <NUM>. In alternative embodiments, the receptacle <NUM> is sized to hold multiple (e.g., eight) specimen containers <NUM>, such that the receptacle <NUM> has an internal diameter of about <NUM> to about <NUM>.

The receptacle <NUM> is typically made of metal. The base housing <NUM> and the lid <NUM> are typically made of materials that provide a degree of thermal insulation, such as polymers. In some embodiments, the cooler <NUM> has a length of about <NUM> to about <NUM>, a height of about <NUM> to about <NUM>, a width of about <NUM> to about <NUM>, and a wall thickness of about <NUM> to about <NUM>. The cooler <NUM> is typically made of one or more insulative materials, such as extruded polystyrene foam or vacuum and laminate container material constructions.

<FIG> illustrate a method of vitrifying a specimen <NUM> within the specimen container <NUM> using the system console <NUM>. Referring to <FIG>, the proximal closure <NUM> is removed (e.g., pulled or twisted) from the elongate tube <NUM> to allow access to the lumen of the elongate tube <NUM>. Referring to <FIG>, a delivery device <NUM> (e.g., a micropipette, a spatula, or another device) is used to deliver the specimen <NUM> to the lumen through the proximal end <NUM> of the elongate tube <NUM>. The specimen <NUM>, along with a small surrounding volume of culture media, may be deposited directly into the equilibration solution <NUM> or deposited just atop the equilibration solution <NUM> within the proximal air pocket <NUM>. The proximal closure <NUM> is then reinstalled to the proximal end <NUM> to reseal the elongate tube <NUM>.

Referring again to <FIG>, the lid <NUM> of the system console <NUM> is opened, and the specimen container <NUM>, with the specimen <NUM> contained therein, is then loaded into the receptacle <NUM> of the system console <NUM>, such that, with respect to the spin axis <NUM>, the distal end <NUM> is spaced further from the spin axis <NUM> than is the proximal end <NUM>. Referring again to <FIG>, the lid <NUM> is closed, and the reader component <NUM> of the system console <NUM> detects the presence of the specimen container <NUM>, such that the timer <NUM> is activated to run for a first predetermined exposure period in response to the detection. The specimen container <NUM> is allowed to sit in place (e.g., stationary) in the receptacle <NUM> for the first predetermined exposure period so that the specimen <NUM> can equilibrate in the equilibration solution <NUM>. The first exposure period may range from about <NUM> minutes to about <NUM> minutes, depending on various parameters of typical ART protocols.

During the first exposure period, the equilibration solution <NUM> draws water molecules out from the specimen <NUM> and infuses cryoprotectants into the specimen <NUM> according to osmotic potential. The reduction of water content and addition of cryoprotectants aids in minimizing damage to cellular components of the specimen <NUM> during freeze and warming cycles. Although the specimen <NUM> is denser than the equilibration solution <NUM> and will therefore very gradually descend through the equilibration solution <NUM> due to gravitational forces over time, the specimen <NUM> will typically still be suspended within the equilibration solution <NUM> and will not have yet reached the separation fluid <NUM> by the end of the first exposure period, as shown in <FIG>.

Referring to <FIG>, once the specimen <NUM> has been exposed to the equilibration solution <NUM> for the predetermined exposure period, the receptacle <NUM> is activated to spin the specimen container <NUM> at a select low speed to advance the equilibration solution <NUM> and the specimen <NUM> axially through the separation fluid <NUM> to the vitrification solution <NUM>. The specimen container <NUM> is typically spun for about <NUM> minutes to about <NUM> minutes at an angular speed of about <NUM> rpm to about <NUM> rpm, which exerts enough centripetal force on the specimen <NUM> to cause the specimen <NUM> to descend into the vitrification solution <NUM> in a timely manner, but not enough to cause mechanical damage to the specimen <NUM>. Such speed (e.g., corresponding to about <NUM> to about <NUM>) is significantly slower than speeds of even very low-speed conventional laboratory centrifuges, which are typically capable of revolving specimens about a centrifuge axis at speeds in a range of about <NUM> rpm to about <NUM>,<NUM> rpm (e.g., corresponding to about <NUM>,<NUM> to about <NUM>,<NUM>).

Referring particularly to <FIG>, during an initial phase of spinning, the specimen <NUM> descends within the equilibration solution <NUM> while the equilibration solution <NUM>, containing the specimen <NUM>, descends via bulk motion through the separation fluid <NUM> (e.g., thereby displacing the separation fluid <NUM>) toward the vitrification solution <NUM>. Referring particularly to <FIG>, during a subsequent phase of spinning, the equilibration solution <NUM> reaches the vitrification solution <NUM>, and the specimen <NUM> passes from the equilibration solution <NUM> into the vitrification solution <NUM>. Referring particularly to <FIG>, during a next phase of spinning, the equilibration solution <NUM> merges with the vitrification solution <NUM> to form a combined vitrification solution <NUM> (e.g., including the equilibration solution <NUM>, the vitrification solution <NUM>, and a mixed solution interface layer between the equilibration solution <NUM> and the vitrification solution <NUM>), and the specimen <NUM> continues to descend through the combined vitrification solution <NUM>.

Referring particularly to <FIG>, during a final phase of spinning, the specimen <NUM> rests on a meniscus <NUM> of the distal air pocket <NUM> due to surface tension and thereby avoids contact with the relatively hard wall of the elongate tube <NUM>. For example, due to a balance between surface tension at the interface of the combined vitrification solution <NUM> and the distal air pocket <NUM>, and tension between combined vitrification solution <NUM> and an interior wall of the tapered portion <NUM>, the potential buoyancy force of the distal air pocket <NUM> is not sufficient to break through meniscus <NUM>. Therefore, the specimen <NUM> cannot penetrate the meniscus <NUM>.

With the specimen <NUM> resting on the meniscus of the distal air pocket <NUM> upon completion of spinning, the timer <NUM> is activated, and the specimen container <NUM> is allowed to sit in place (e.g., stationary) in the receptacle <NUM> for a second predetermined exposure period for the specimen <NUM> to be exposed to the combined vitrification solution <NUM>. The second exposure period may range from about <NUM> minutes to about <NUM> minutes, depending on various parameters of typical ART protocols. During the second exposure period, permeation of cryoprotectants within the combined vitrification solution <NUM> into the specimen <NUM> replaces water within the specimen <NUM>, thereby dehydrating the specimen and further infusing the specimen <NUM> with cryoprotectants. Such a stage-like progression of media concentrations avoids an excessively high initial osmotic differential that could otherwise cause cells of the specimen <NUM> to shrink too much and too rapidly as the water leaves the cells at a rate faster than the cryoprotectants can enter the cells.

Owing to a preloaded state of the equilibration solution <NUM> and the vitrification solution <NUM> within the specimen container <NUM>, a specimen can be prepared for vitrification within a single, isolated environment (e.g., the lumen of the specimen container <NUM>) without being exposed to contamination, mechanical damage (e.g., from a micropipette or other specimen holding or fluid delivery device), or other accidental mishandling that may otherwise occur when a container that houses a specimen is accessed multiple times to deliver and remove various processing mediums or when a specimen is moved to various containers (e.g., petri dishes, test tubes, or flask) during an ART process.

In some implementations, once the second exposure period has ended, the specimen container <NUM>, containing the specimen <NUM>, is released directly from the receptacle <NUM> downward through the exit channel <NUM> of the base housing <NUM> (refer to <FIG>) and into the low temperature substance (e.g., liquid N<NUM> at a temperature of about - <NUM>) contained within the cooler <NUM> for temporary low temperature storage. The specimen container <NUM> is deposited in a manner such that at least a distal portion of the specimen container <NUM> surrounding the specimen <NUM> is submerged in the low temperature substance. The timer <NUM> is activated, causing the specimen <NUM> to rapidly cool to a glass state before ice crystals can form within cells of the specimen <NUM> so that specimen <NUM> can be preserved in a viable state. The specimen container <NUM>, containing the specimen <NUM>, is then manually transferred from the cooler <NUM> of the system console <NUM> to a long-term low temperature storage structure, where the specimen <NUM> can be maintained in a cryogenic state for a period of up to about <NUM> years. In some instances, the specimen container <NUM> may be stored in the long-term low temperature storage structure for a much shorter period (e.g., as short as few hours).

In some alternative implementations, once the second exposure period has ended, the specimen container <NUM>, containing the specimen <NUM>, is manually removed from the receptacle <NUM>, visually inspected, and then subsequently reinserted into the receptacle <NUM> for release into the cooler <NUM>, as opposed to being immediately released downward into the cooler <NUM> upon termination of the second exposure period. Referring to <FIG>, in some implementations, once the second exposure period has ended, the specimen container <NUM>, containing the specimen <NUM>, is manually removed from the receptacle <NUM> and immersed in a low temperature substance <NUM> within a beaker <NUM> or other container instead of being released into the cooler <NUM>. As discussed above, the specimen container <NUM> is immersed in a manner such that at least a distal portion of the specimen container <NUM> surrounding the specimen <NUM> is submerged in the low temperature substance <NUM>. The timer <NUM> (or another timer) can be activated to track the relatively short duration in which the specimen container <NUM> is submerged. The specimen container <NUM>, containing the specimen <NUM>, is then manually transferred from the beaker <NUM> to a long-term low temperature storage structure.

Referring to <FIG>, the specimen container <NUM> can be stored in the long-term low temperature storage structure until the specimen <NUM> is ready to be used in reproductive or other procedures. At such a time, the specimen container <NUM> can be removed from the storage structure and subsequently thawed via standard warming protocols in which the specimen <NUM> is exposed to one or more warming solutions. For example, referring to <FIG>, the specimen container <NUM>, containing the specimen <NUM>, is transferred to a one or more warming solutions <NUM> at a temperature of typically about <NUM> for a period of about <NUM> seconds to about <NUM> minute. In some implementations, the warming solutions <NUM> may be at about room temperature. Referring to <FIG>, the specimen container <NUM> is opened by one or both of removing the distal closure <NUM> (e.g., cutting off the distal closure <NUM> along the distal air pocket <NUM>) from the elongate tube <NUM> (<FIG>) and removing (e.g., pulling, twisting, or cutting) the proximal closure <NUM> from the elongate tube (<FIG>). Referring to <FIG>, the specimen <NUM> and the combined vitrification solution <NUM> can then be dispelled (e.g., drained or purged) from the opened specimen container <NUM> into a petri dish <NUM> or other container at a temperature of about <NUM> for further processing of the specimen <NUM> according to selected ART protocols.

While certain embodiments of specimen containers have been described above, other embodiments are possible.

While certain implementations of vitrifying a specimen have been described above, other implementations are possible. For example, while the process for vitrifying the specimen <NUM> has been described as including the step of immersing the specimen container <NUM> within a low temperature substance following spinning within the console <NUM>, in some implementations, the specimen <NUM> is released onto a conventional specimen carrier for immediate use in an ART procedure without exposing the specimen container <NUM> to the low temperature substance for cryopreservation. In such cases, the specimen container <NUM> is discarded following release of the specimen <NUM>.

While the process for vitrifying the specimen <NUM> has been described as including the step of spinning the specimen container <NUM> within the console <NUM> to gradually sediment the specimen <NUM> through the equilibration solution <NUM> and the vitrification solution <NUM>, in some implementations, the specimen <NUM> can be grasped manually with an appropriate tool (e.g., a micropipette), manually immersed in the equilibration solution <NUM> for a defined period of time with the tool, advanced with the tool into the vitrification solution <NUM>, and held in the vitrification solution <NUM> with the tool for a defined period of time. The specimen <NUM> is then released from the tool into the vitrification solution <NUM> for exposure of the specimen container <NUM>, with the specimen <NUM> contained therein, to a low temperature substance, or the specimen <NUM> is manually withdrawn from the elongate tube <NUM> with the tool and submerged directly into liquid nitrogen or another cooling substance with the tool.

While the process for vitrifying the specimen <NUM> has been described as including the step of spinning the specimen container <NUM> within the receptacle <NUM> of the console <NUM> about the spin axis <NUM>, in some implementations, the specimen container <NUM>, with the specimen <NUM> contained therein, may be revolved within a conventional centrifuge that is designed to revolve specimen containers at appropriately low speeds about the centrifuge axis.

While the process for vitrifying the specimen <NUM> has been described as vitrifying a single specimen <NUM> at a time within the specimen container <NUM>, in some implementations, multiple specimens may be deposited into a single specimen container <NUM> for simultaneous processing according to the process described above with respect to <FIG>. In some embodiments, multiple specimen containers <NUM>, each carrying one or more specimens <NUM> may be placed within the receptacle <NUM> together for simultaneous spinning.

While the specimen container <NUM> has been described as including the proximal closure <NUM> formed as a cap that surrounds an exterior wall of the elongate tube <NUM>, in some embodiments, a specimen container <NUM> includes a proximal closure <NUM> formed as a plug (e.g., a cork) that seats within the lumen of the elongate tube <NUM>, as shown in <FIG>. While some features have been omitted from the drawing for clarity, the specimen container <NUM> is substantially similar in construction and function to the specimen container <NUM>, except that the specimen container <NUM> includes the proximal closure <NUM> instead of the proximal closure <NUM>. The proximal closure <NUM> includes a plug <NUM> (e.g., an inserting portion) that is sized to be snuggly inserted into the lumen of the elongate tube <NUM> to seal the elongate tube <NUM> at the proximal end <NUM>. The proximal closure <NUM> further includes a top flange <NUM> that is wider than the elongate tube <NUM> such that the top flange <NUM> remains external to the lumen while the plug <NUM> is disposed within the lumen to facilitate handling the proximal closure <NUM>. The proximal closure <NUM> is typically made of plastic.

In some embodiments, a specimen container includes a proximal closure that has a specimen carrying portion. <FIG> illustrates a specimen container <NUM> that includes such a feature. The specimen container <NUM> is substantially similar in construction and function to the specimen container <NUM>, except that the specimen container <NUM> includes a closure support <NUM> and a proximal closure <NUM> instead of the proximal closure <NUM>. The closure support <NUM> is formed as a circumferential leaf structure that is wider than the elongate tube <NUM> and that lies adjacent the proximal end <NUM> of the elongate tube <NUM>. The proximal closure <NUM> includes a plug <NUM> that is sized to be snuggly inserted into a tubular portion <NUM> of the closure support <NUM> and to rest against an annular platform <NUM> of the closure support <NUM> to seal the elongate tube <NUM> at the closure support <NUM>. The plug <NUM> is sized to extend past an end of the closure support <NUM> when resting on the annular platform <NUM> such that the plug <NUM> can be used to manipulate the proximal closure <NUM>. The proximal closure <NUM> further includes a specimen carrier <NUM> that is sized to hold a specimen <NUM> and to deliver the specimen <NUM> to the lumen of the elongate tube <NUM>. The delivery, or transfer, of specimen <NUM> into equilibration solution <NUM> take may place by simple immersion, or g-forces could be applied to the specimen <NUM> to urge the specimen <NUM> off of the specimen carrier <NUM> into the equilibration solution <NUM>. The proximal closure <NUM> is typically made of plastic.

In some embodiments, a specimen container that is similar to any of the specimen containers <NUM>, <NUM>, <NUM> described above or any of the specimen containers described below includes a distal closure that is formed as a plug.

While the specimen container <NUM> has been described as including a separation <NUM> fluid (e.g., an air bubble) that separates the equilibration solution <NUM> from the vitrification solution <NUM>, in some embodiments, a specimen container includes a different separation mechanism. For example, <FIG> illustrates a portion of a specimen container <NUM> that includes a mechanical separation member <NUM> that serves as a barrier between the equilibration solution <NUM> and the vitrification solution <NUM>. The specimen container <NUM> is substantially similar in construction and function to the specimen container <NUM>, except that the specimen container <NUM> includes the mechanical separation member <NUM> instead of the separation fluid <NUM>. In the example embodiment <NUM>, the mechanical separation member <NUM> is provided as a barrier (e.g., a butterfly valve) that prevents passage of water vapor between, as well as prevents premature mixing of, the equilibration solution <NUM> and the vitrification solution <NUM> The mechanical separation member <NUM> is designed to remain closed under nominal conditions and to open under sufficient g-force loading, as will occur during spinning of the specimen container <NUM> within the system console <NUM> or within an appropriately designed centrifuge.

Other examples of mechanical separation members that may be included in similar specimen containers include a sphere that fits snugly within a local internal constriction, a separation fluid with properties that promote desired migration of the separation fluid while under centripetal loading (e.g., a separation fluid with a density that is less than that of the equilibration solution <NUM>), a viscoelastic fluid that moves only when subject to sufficiently high g-force, and a film that can be pierced (or otherwise penetrated) to create a pathway large enough for the specimen <NUM> to easily pass through the film.

While the specimen container <NUM> has been described as including one equilibration solution <NUM> and one vitrification solution <NUM>, in some embodiments, a specimen container includes more than one equilibration solution and/or more than one vitrification solution <NUM>. For example, <FIG> illustrates a portion of a specimen container <NUM> that includes multiple vitrification solutions. The specimen container <NUM> is substantially similar in construction and function to the specimen container <NUM>, except that the specimen container <NUM> includes the multiple vitrification solutions <NUM>, <NUM>, <NUM> of different densities, which may be useful in situations where it is advantageous to process a specimen <NUM> in smaller graduations of concentration. In some embodiments, the specimen container <NUM> is preloaded and packaged with one initial vitrification solution <NUM>, which then separates into the three vitrification solutions <NUM>, <NUM>, <NUM> upon the specimen container <NUM> being removed from packaging and subjected to high g-forces (e.g., about <NUM>,<NUM>) for a short period of time (e.g., about <NUM> seconds). In this manner, a concentration gradient of the vitrification solutions <NUM>, <NUM>, <NUM> can be created just prior to insertion of a specimen into the specimen container <NUM>.

In some embodiments, a specimen container includes one or both of an equilibration solution and a vitrification solution with magnetic properties. For example, <FIG> illustrate a portion of a specimen container <NUM> with such a feature. The specimen container <NUM> is substantially similar in construction and function to the specimen container <NUM>, except that the specimen container <NUM> includes an equilibration solution <NUM> that is loaded with magnetic nanoparticles <NUM> formed of iron oxide (Fe<NUM>O<NUM>). The magnetic nanoparticles <NUM> are coated with a biocompatible, inert substance, such as biotin or polyethylene glycol (PEG).

Referring particularly to <FIG>, during a vitrification process, a specimen <NUM> can be delivered to the equilibration solution <NUM> as described above with respect to <FIG>. Referring to <FIG>, an external magnetic field source <NUM> can then be turned on to provide a constant (e.g., non-alternating) magnetic field that pulls the magnetic nanoparticles <NUM> and the surrounding equilibration solution <NUM> downward into the vitrification solution <NUM> to form a combined vitrification solution <NUM>. In some embodiments, the magnetic field source <NUM> is provided as a case that is designed to contain the specimen container <NUM>. Accordingly, the magnetic field source <NUM> may be designed in a manner so as to act on the specimen container <NUM> while shielding other surrounding objects from the magnetic field. Downward movement of the magnetic nanoparticles <NUM> and the surrounding equilibration solution <NUM> in turn drags the specimen <NUM> downward toward a meniscus <NUM> of the combined vitrification solution <NUM>. Owing to the sedimentation of the specimen <NUM> by a magnetic field, the specimen container <NUM> may not need to undergo a spinning process, as discussed above with respect to the system console <NUM>.

While the specimen container <NUM> has been described as including an ID label <NUM> in the form of an RFID tag, in some embodiments, a specimen container includes an ID label in the form of a barcode or a quick response (QR) code. For example, <FIG> respectively illustrate portions of specimen containers <NUM>, <NUM> that include ID labels <NUM>, <NUM> in the form of a barcode and a QR code at the proximal end <NUM> of the elongate tube <NUM>. The specimen containers <NUM>, <NUM> are otherwise substantially similar in construction and function to the specimen container <NUM>.

In some embodiments, as shown in <FIG>, an ID label may, itself, serve as a proximal closure (e.g., having a sterile internal surface) and may therefore be provided as a part of a specimen container in lieu of a cap-like or plug-like proximal closure. For example, the specimen container <NUM> includes an ID label <NUM> in the form of an RFID tag and that is coated with an adhesive. <FIG> illustrates the ID label <NUM> in an open configuration, while <FIG> illustrates the ID label <NUM> in a wrapped, closed configuration. The ID label <NUM> includes a printable region <NUM> on which a user can write on an outer surface. The specimen container <NUM> is substantially similar in construction and function to the specimen container <NUM>, except that the specimen container <NUM> includes the ID label <NUM> in a configuration that serves as a proximal closure of the elongate tube <NUM>.

In some embodiments, as shown in <FIG>, a specimen container <NUM> includes an elongate tube <NUM> with a flared proximal end <NUM>. The specimen container <NUM> is substantially similar in construction and function to the specimen container <NUM> except that the specimen container <NUM> includes the elongate tube <NUM> with the flared proximal end <NUM> instead of the elongate tube <NUM> with the tubular proximal end <NUM>. Though omitted for clarity, the specimen container <NUM> further includes the equilibration solution <NUM>, the separation fluid <NUM>, the vitrification solution <NUM>, the proximal air pocket <NUM>, and the distal air pocket <NUM>. The flared proximal <NUM> end is formed as a receptacle that is wider than the elongate tube <NUM>, thereby facilitating loading of a specimen within a lumen of the elongate tube <NUM> using a delivery device <NUM>. In some embodiments, a similar specimen container includes an elongate tube with a flared proximal end of a shape different from that shown in <FIG>.

In some embodiments, a specimen container includes a bulbous region that acts as a bulb syringe to aid in dispelling a specimen from the specimen container without opening both ends of the specimen container. For example, <FIG> respectively illustrate specimen containers <NUM>, <NUM>, <NUM> that include bulbous regions <NUM>, <NUM>, <NUM> located at proximal, distal, and central regions of the specimen containers <NUM>, <NUM>, <NUM>. The specimen containers <NUM>, <NUM>, <NUM> are substantially similar in construction and function to the specimen container <NUM>, except that the specimen containers <NUM>, <NUM>, <NUM> include the bulbous regions <NUM>, <NUM>, <NUM> along elongate tubes <NUM>, <NUM>, <NUM>. In some embodiments, a vision system may be used to view a state and position of a specimen as the specimen is dispelled from a specimen container <NUM>, <NUM>, <NUM>. For example, a re-expansion state of the specimen after residing in equilibration solution <NUM> would indicate a state of osmotic equilibration, which could indicate to the system that the specimen is ready to advance to the vitrification solution <NUM>).

In some embodiments, a specimen container includes an access port for depositing a specimen into a lumen of the specimen container. For example, <FIG> illustrates a specimen container <NUM> including an elongate tube <NUM> that defines an access port <NUM>. The specimen container <NUM> further includes a proximal closure <NUM> that hermetically seals a proximal end region of the elongate tube <NUM> and a distal closure <NUM> that hermetically seals a distal end <NUM> of the elongate tube <NUM>. The elongate tube <NUM> is a thin capillary tube of very small diameter (e.g., having an internal diameter on the order of <NUM>-<NUM> m). The elongate tube <NUM> has a substantially constant diameter along a main portion <NUM> (e.g., a cylindrical portion) and has a variable diameter that gradually decreases along a tapered portion <NUM> that extends from the main portion <NUM> to the distal end <NUM>.

The proximal closure <NUM> is a plunger that is designed to seat within a lumen of the elongate tube <NUM> to close a proximal end <NUM> and the access port <NUM> of the elongate tube <NUM>. Accordingly, the proximal closure <NUM> includes a plug <NUM> (e.g., an elongate cylindrical member) that is sized to be inserted into the lumen of the elongate tube <NUM> and a grasping member <NUM> that abuts the proximal end <NUM> of the elongate tube <NUM> (e.g., thereby remaining external to the lumen) when the plug <NUM> is appropriately disposed within the lumen. The grasping member <NUM> may have a smooth (e.g., cylindrical) outer surface or a faceted (e.g., hexagonal) outer surface that facilitates handling of the proximal closure <NUM> and that limits movement of the proximal closure <NUM> in instances when the proximal closure <NUM> is separated from the elongate tube <NUM> and placed atop a surface. In some embodiments, the exterior surface of the grasping member <NUM> may have an asymmetric profile to prevent such undesired movement atop a surface. The proximal closure <NUM> can be reversibly installed and removed from the proximal end region of the elongate tube <NUM> to seal the proximal end <NUM> and the access port <NUM> and to open the proximal end region to allow proximal access to the elongate tube <NUM> via the access port <NUM>, respectively. The distal closure <NUM> is a single-use seal (e.g., a melt seal, a fold, glue or adhesive, or another occluding member) that can be removed (e.g., cut or otherwise separated) from the distal end <NUM> of the elongate tube <NUM> to allow material to pass distally out of the elongate tube <NUM>.

The main portion <NUM> of the elongate tube <NUM> typically has a length of about <NUM> to about <NUM> (e.g., about <NUM>), an outer diameter of about <NUM> to about <NUM> (e.g., about <NUM>), and a wall thickness of about <NUM> to about <NUM> (e.g., about <NUM>). The tapered portion <NUM> of the elongate tube <NUM> typically has a length of about <NUM> to about <NUM> (e.g., about <NUM>), a maximum outer diameter that is adjacent and equal to the outer diameter of the main portion <NUM>, a minimum outer diameter (e.g., at the distal end <NUM> of the elongate tube <NUM>) of about <NUM> to about <NUM> (e.g., about <NUM>), and a wall thickness of about <NUM> to about <NUM> (e.g., about <NUM>). The proximal closure <NUM> has a length of about <NUM> to about <NUM> (e.g., about <NUM>).

The access port <NUM> has an elliptical cross-sectional shape and has a width that is about equal to the diameter of the main portion <NUM>. The access port <NUM> typically has a length of about1 mm to about <NUM> (e.g., about <NUM>). A center of the access port <NUM> is typically located about <NUM> to about <NUM> (e.g., about <NUM>) from the proximal end <NUM> of the elongate tube <NUM>. A lumen of the elongate tube <NUM>, at a smallest inner diameter, is large enough to accommodate a specimen. A geometry and a construction (e.g., a thin and small profile) of the elongate tube <NUM> are configured to maximize heat transfer and to minimize thermal mass to maximize cooling and warming rates of the specimen container <NUM> during ART protocols. The specimen container <NUM> typically has a total length (e.g., including lengths of the elongate tube <NUM>, the proximal closure <NUM> as installed, and the distal closure <NUM>) of about <NUM> to about <NUM> (e.g., about <NUM>).

The proximal closure <NUM> may be made of one or materials, including plastic and stainless steel. The elongate tube <NUM> may be manufactured according to the processes discussed above with respect to the elongate tube <NUM> and formed from the same materials as those of the elongate tube <NUM>, as discussed above. As further discussed with respect to the specimen container <NUM>, the lumen of the elongate tube <NUM> is internally preloaded with multiple fluids (omitted from <FIG> for clarity) located distal to the access port <NUM>, sequentially including the equilibration solution <NUM>, the vitrification solution <NUM>, the separation fluid <NUM>, and the distal air pocket <NUM> in volumetric amounts discussed above. The specimen container <NUM> further includes the ID label <NUM> (omitted for clarity) attached to the elongate tube <NUM> near the proximal end <NUM>.

Similar to the specimen container <NUM>, the specimen container <NUM> is a sterile, single-use device that is non-toxic to specimens contained therein. The specimen container <NUM> may be packaged as a single unit, and both the specimen container <NUM> and the packaging will remain sterile for a guaranteed shelf-life of the specimen container <NUM>. The total length of the specimen container <NUM> typically allows the specimen container <NUM> to fit within standard storage containers and other standard equipment used in ART protocols.

During a process of vitrifying a specimen <NUM> within the specimen container <NUM>, the proximal closure <NUM> is removed (e.g., pulled or twisted) from the elongate tube <NUM> to open the access port <NUM>. A delivery device (e.g., such as the delivery device <NUM>) is used to deliver the specimen <NUM>, suspended within a small amount of culture media, to the lumen of the elongate tube <NUM> through the access port <NUM>. The specimen <NUM> and the culture media may be deposited directly into the equilibration solution <NUM> or deposited just proximal to the equilibration solution <NUM>. The proximal closure <NUM> is then reinstalled to the elongate tube <NUM> to reseal the proximal end <NUM> and the access port <NUM> for further processing of the specimen <NUM>.

In some embodiments, a handle can be used to house, store, and manipulate the specimen container <NUM>. For example, <FIG> illustrates such a handle <NUM>. The handle <NUM> includes a handle body <NUM> and a cap <NUM> that is formed to close the handle body <NUM>. The handle body <NUM> is open at a proximal end <NUM> and is sized to carry the specimen container <NUM>. The handle body <NUM> includes a main portion <NUM> that surrounds the main portion <NUM> of the elongate tube <NUM> and a distal support <NUM> that supports the tapered portion <NUM> of the elongate tube <NUM>.

The main portion <NUM> has a faceted (e.g., hexagonal) exterior surface and cross-sectional shape that facilitates handling of the handle body <NUM> and that prevents rolling or other undesired movement of handle <NUM> atop a surface. In some embodiments, the exterior surface of the main portion <NUM> may have an asymmetric profile to prevent such undesired movement atop a surface. The main portion <NUM> defines a window <NUM> that aligns with the access port <NUM> when the elongate tube <NUM> is securely disposed within the handle <NUM>. The window <NUM> is an opening defined by a cutout in the main portion <NUM> of the handle body <NUM>. The plug <NUM> is slidably guided within the handle body <NUM> and may be biased to move, thereby occluding the access port <NUM>, by g-forces induced as would occur during spinning of the specimen container <NUM>, integrated inside of handle <NUM>, within the system console <NUM> or within an appropriately designed centrifuge The distal support <NUM> is formed as a cylindrical tube that extends from the main portion <NUM> to support the tapered portion <NUM> of the elongate tube <NUM> that passes therethrough. The cap <NUM> is formed as a tubular member that can seat against the main portion <NUM> and the distal support <NUM> via a friction fit to distally close the handle body <NUM>. The cap <NUM> can be removed from the handle body <NUM> to allow access to the proximal end <NUM> of the specimen container <NUM>.

The main portion <NUM> of the handle body <NUM> typically has a length of about <NUM> to about <NUM> (e.g., about <NUM>), a width of about <NUM> to about <NUM> (e.g., about <NUM>), and a wall thickness of about <NUM> to about <NUM> (e.g., about <NUM>). The window <NUM> typically has a length of about <NUM> to about <NUM> (e.g., about <NUM>). A center of the window <NUM> is typically located about <NUM> to about <NUM> (e.g., about <NUM>) from the proximal end <NUM> of the handle body <NUM>. Although the main portion <NUM> has a hexagonal exterior cross-sectional shape, the main portion <NUM> has a cylindrical interior cross-sectional shape for smooth housing of the specimen container <NUM>. The distal guide <NUM> of the handle body <NUM> typically has a length of about <NUM> to about <NUM> (e.g., about <NUM>), and an outer diameter of about <NUM> to about <NUM> (e.g., about <NUM>). The tapered portion <NUM> of the specimen container <NUM> typically extends about <NUM> to about <NUM> past the distal guide <NUM> when the specimen container <NUM> is secured within the handle <NUM>. The cap <NUM> of the handle <NUM> typically has a length of about <NUM> to about <NUM> (e.g., about <NUM>), a diameter of about <NUM> to about <NUM> (e.g., about <NUM>), and a wall thickness of about <NUM> to about <NUM> (e.g., about <NUM>). The handle body <NUM> is typically manufactured via injection molding and may be made of one or more materials, such as polystyrene, polypropylene, polycarbonate, and other injection moldable resins. The cap <NUM> is typically manufactured via injection molding or extrusion and may be made of one or more materials, such as polystyrene, polypropylene, polycarbonate, and other injection moldable or extrusion grade resins.

In some embodiments, the handle <NUM> or a handle that is substantially similar in construction and function to the handle <NUM> (e.g., but without the window <NUM>) can be used to house, store, and manipulate any of the above-discussed specimen containers.

In some embodiments, a specimen container includes a sieve for draining media from a distal end of the specimen container. For example, <FIG> illustrate a specimen container <NUM> including a sieve <NUM>. The specimen container <NUM> further includes an elongate tube <NUM>, a proximal sharp tube <NUM> that joins at the proximal end region of the elongate tube <NUM> and a distal closure <NUM> that hermetically seals a distal end <NUM> of the elongate tube <NUM>. The elongate tube <NUM> is a thin capillary tube of very small diameter (e.g., having an internal diameter on the order of <NUM>-<NUM> m). The elongate tube <NUM> has a substantially constant diameter along a main portion <NUM> (e.g., a cylindrical portion) and has a variable diameter that gradually decreases along a tapered portion <NUM> that extends from the main portion <NUM> to the distal end <NUM>.

The proximal member <NUM> is a sharp tube with a blunt distal end and sharp proximal end. The proximal member <NUM> is designed to slidably mate within a lumen of the elongate tube <NUM> to seal the inner diameter of tube <NUM> at the proximal end <NUM> and to seal the access port <NUM> of the elongate tube <NUM>. A vial <NUM> (e.g., a vessel) contains the equilibration solution <NUM>. During an initial centrifuge spin, the vial <NUM> can be advanced to be pierced on the proximal member <NUM> and to drain. The distal closure <NUM> is a single-use seal (e.g., a melt seal, a fold, glue or adhesive, or another occluding member) that can be removed (e.g., cut or otherwise separated) from the distal end <NUM> of the elongate tube <NUM> to allow material to pass distally out of the elongate tube <NUM>.

A delivery device (e.g., such as a micropipette) may be used to deliver the specimen, suspended within a small amount of culture media, to the lumen of the elongate tube <NUM> through the access port <NUM>. The specimen and a small volume of culture media may be deposited directly into the inner lumen of the elongate tube <NUM>.

The vial <NUM> is biased (e.g., by centripetal force or by pushing) until the sharp tube <NUM> pierces a frangible distal membrane of the vial <NUM>. Simultaneously, the same forces acting on the vial <NUM> slide the sharp tube <NUM> distally within the elongate tube <NUM> until the access port <NUM> is occluded by the distal, blunt, end of sharp tube <NUM>, thereby trapping the specimen and culture media. Upon piercing, the contents (e.g., equilibration solution <NUM>) of the vial <NUM> may drain through the lumen of sharp tube <NUM> and into the closed internal volume of elongate tube <NUM>.

The sieve <NUM> is a collection of holes <NUM> (e.g., laser-drilled holes) that are arranged in multiple rows along the tapered portion <NUM> of the elongate tube <NUM>. The sieve <NUM> allows fluids (e.g., culture media, equilibration solution, vitrification solution, and other excess fluids) to drain distally from the specimen container <NUM> during a spinning phase of a vitrification procedure, while a specimen is retained within the tapered portion <NUM> of the elongate tube <NUM> distal to the sieve <NUM>. The spinning phase of the vitrification procedure drains the excess fluid until the point where the specimen resides either within the sieve <NUM> or slightly distal to the sieve in the vitrification solution <NUM> of the tapered portion <NUM>. The sieve <NUM> typically has a total length of about <NUM> to about <NUM> and is spaced about <NUM> to about <NUM> from the distal end <NUM> of the elongate tube <NUM>. Rows of the holes <NUM> are typically spaced apart by about <NUM> to about <NUM> from each other, each hole <NUM> typically has a diameter of about <NUM> to about <NUM>.

The main portion <NUM> of the elongate tube <NUM> typically has a length of about <NUM> to about <NUM> (e.g., about <NUM>), an outer diameter of about <NUM> to about <NUM> (e.g., about <NUM>), and a wall thickness of about <NUM> to about <NUM> (e.g., about <NUM>). The tapered portion <NUM> of the elongate tube <NUM> typically has a length of about <NUM> to about <NUM> (e.g., about <NUM>), a maximum outer diameter that is adjacent and equal to the outer diameter of the main portion <NUM>, a minimum outer diameter (e.g., at the distal end <NUM> of the elongate tube <NUM>) of about <NUM> to about <NUM> (e.g., about <NUM>), and a wall thickness of about <NUM> to about <NUM> (e.g., about <NUM>). The proximal member <NUM> has a length of about <NUM> to about <NUM> (e.g., about <NUM>).

The access port <NUM> has an elliptical cross-sectional shape and has a width that is about equal to the diameter of the main portion <NUM>. The access port <NUM> typically has a length of about <NUM> to about <NUM> (e.g., about <NUM>). A center of the access port <NUM> is typically located about <NUM> to about <NUM> (e.g., about <NUM>) from the proximal end <NUM> of the elongate tube <NUM>. A lumen of the elongate tube <NUM>, at a smallest inner diameter, is large enough to accommodate a specimen. A geometry and a construction (e.g., a thin and small profile) of the elongate tube <NUM> are configured to maximize heat transfer and to minimize thermal mass to maximize cooling and warming rates of the specimen container <NUM> during ART protocols. The specimen container <NUM> typically has a total length (e.g., including lengths of the elongate tube <NUM> and the distal closure <NUM>) of about <NUM> to about <NUM> (e.g., about <NUM>).

The proximal closure <NUM> may be made of one or materials, including stainless steel hypodermic tubing. The elongate tube <NUM> may be manufactured according to the processes discussed above with respect to the elongate tube <NUM> and formed from the same materials as those of the elongate tube <NUM>, as discussed above. As further discussed with respect to the specimen container <NUM>, the lumen of the elongate tube <NUM> is internally preloaded with multiple fluids (omitted from <FIG> for clarity) located distal to the access port <NUM>, sequentially ,starting adjacent to the distal end <NUM> and moving proximally, the vitrification solution <NUM>, and a separation fluid <NUM>. The specimen container <NUM> further includes the ID label <NUM> (omitted for clarity) attached to the elongate tube <NUM> near the proximal end <NUM>.

Similar to the specimen container <NUM>, the specimen container <NUM> is a sterile, single-use device that is non-toxic to specimens contained therein. The specimen container <NUM> may be individually packaged, and both the specimen container <NUM> and the packaging will remain sterile for a guaranteed shelf-life of the specimen container <NUM>. The total length of the specimen container <NUM> typically allows the specimen container <NUM> to fit within standard storage containers and other standard equipment used in ART protocols.

In some embodiments, a handle can be used to house, store, and manipulate the specimen container <NUM>. For example, <FIG> illustrates such a handle <NUM>. The handle <NUM> is substantially similar in construction to the handle <NUM>, except that a window <NUM> of the handle is sized and positioned to align with the access port <NUM> of the specimen container <NUM>. Accordingly, the handle <NUM> includes a handle body <NUM> and a cap that is formed to close the handle body <NUM>.

While the system console <NUM> has been described as including a reader component <NUM> that is positioned and programmed to detect a presence of the specimen container <NUM> within the receptacle <NUM>, in some embodiments, a system console that is substantially similar to the system console <NUM> includes an additional reader component that is programmed and positioned at a suitable location to read the ID label <NUM> to detect a presence of the specimen container <NUM> within the cooler <NUM>.

While the system console <NUM> has been described as including a single receptacle <NUM>, in some embodiments, a system console that is substantially similar to the system console <NUM> includes multiple receptacles <NUM> that can each receive a specimen container and simultaneously spin multiple specimen containers.

While the specimen container <NUM> has been described and illustrated as including the separation fluid <NUM> as a barrier between the equilibrium solution <NUM> and the vitrification solution <NUM> within the specimen container <NUM>, in some embodiments, a specimen container includes a different type of barrier for separating solutions within the container. For example, <FIG> illustrate a main portion <NUM> of a specimen container <NUM> that includes a different type of separation barrier.

The specimen container <NUM> is similar in function and in some respects similar in construction to the specimen container <NUM>. For example, as discussed above with respect to the specimen container <NUM>, the specimen container <NUM> can be used to prepare a specimen <NUM> according to a biological or other protocol and to subsequently store the specimen <NUM> in a low temperature substance in a viable and vitrified state. Furthermore, the specimen container <NUM> is a sterile, single-use device that is non-toxic to specimens contained therein. The specimen container <NUM> may be packaged as a single unit, and both the specimen container <NUM> and the packaging will remain sterile for a guaranteed shelf-life of the specimen container <NUM>. A total length of the specimen container <NUM> typically allows the specimen container <NUM> to fit within standard storage containers and other standard equipment used in ART protocols.

The specimen container <NUM> includes an elongate tube <NUM> and two clips <NUM> (e.g., mechanical crimping members) that are attached to the elongate tube <NUM>. The clips <NUM> are attached at different positions to separate an equilibration solution <NUM>, a first vitrification solution <NUM>, and a second vitrification solution <NUM> from each other within the elongate tube <NUM>. The elongate tube <NUM> is a flexible member, and the clips <NUM> are biased to a closed configuration during manufacturing of the specimen container <NUM> such that the clips <NUM> squeeze the elongate tube <NUM> closed at selected locations to form hermetically sealed chambers <NUM>, <NUM>, <NUM> that respectively contain the equilibration solution <NUM>, the first vitrification solution <NUM>, and the second vitrification solution <NUM>, as shown in <FIG>.

For example, the clips <NUM> are sequentially closed as the solutions <NUM>, <NUM>, <NUM> are sequentially delivered to the elongate tube <NUM>. The clips <NUM> are oriented perpendicular (e.g., transverse) to an axis <NUM> of the elongate tube <NUM>. The specimen container <NUM> also includes the ID label <NUM> (refer to <FIG>) at a proximal end and includes proximal and distal closures that respectively, hermetically seal the proximal and distal ends (not shown) of the elongate tube <NUM>, as discussed above with respect to like components of the specimen container <NUM>. The volumes, properties, and constituencies of the solutions <NUM> and <NUM>, <NUM> within the specimen container <NUM> are equivalent to those of the solutions <NUM> and <NUM> discussed above with respect to the specimen container <NUM>.

As discussed above with respect to the elongate tube <NUM>, the elongate tube <NUM> is a thin capillary tube of very small diameter and has a substantially constant diameter along the main portion <NUM>. The elongate tube <NUM> has a variable diameter that gradually decreases along a tapered portion (e.g., similar to the tapered portion <NUM>) that extends from the main portion <NUM> to the distal end. The elongate tube <NUM> is also sized as described above with respect to the elongate tube <NUM> and may be manufactured via plastic extrusion or other techniques and secondary manufacturing treatments (e.g., thermal stretching and reduction) to achieve the desired geometry. The elongate tube <NUM> is typically made of one or more semi-elastic materials that can withstand the low temperature substance, including but not limited to polyolefins, polycarbonates, and polystyrenes. The elongate tube <NUM> is also typically transparent or translucent to allow viewing of the specimen <NUM> through the wall of the elongate tube <NUM>. The clips <NUM> are typically made of one or more corrosion resistant and malleable materials that can suitably deform for closing and opening, such as stainless steel and aluminum.

During a process for vitrifying the specimen <NUM> within the specimen container <NUM>, the proximal closure is removed from the elongate tube <NUM>, and a delivery device (e.g., such as the delivery device <NUM>) is used to deliver the specimen <NUM>, suspended within a small amount of culture media to the equilibration solution <NUM> through a proximal opening of the elongate tube <NUM>. The proximal closure is then reinstalled to the elongate tube <NUM> to reseal the proximal opening for further processing of the specimen <NUM> within a system console.

A system console in which the specimen container <NUM> is disposed can be controlled to bias the clips <NUM> to an open configuration (refer to <FIG>) at the same time or in a timed sequence. The open configuration of a clip <NUM> opens a lumen of the elongate tube <NUM> to permit distal movement of the solutions <NUM>, <NUM>, <NUM> and the specimen <NUM> (e.g., with minimal friction along an inner surface of the elongate tube <NUM>) according to a specimen processing protocol in which g-forces are applied to the specimen container <NUM>. In some embodiments, the clips <NUM> may be designed to be re-closeable for constraining a fluid volume within the elongate tube <NUM> during or after processing of the specimen <NUM> within the specimen container <NUM>.

While the specimen container <NUM> has been described and illustrated as including the transversely oriented clips <NUM>, in some embodiments, a specimen container that is otherwise substantially similar in construction and function to the specimen container <NUM> includes clips that are oriented parallel to an elongate tube of the specimen container. For example, <FIG> illustrates a specimen container <NUM> including the elongate tube <NUM> and clips <NUM> that are oriented parallel to the axis <NUM> of the elongate tube <NUM>. The specimen container <NUM> is otherwise substantially similar in construction, function, and manner of use to the specimen container <NUM>, and the clips <NUM> have the same material formulation as that of the clips <NUM>.

While the specimen container <NUM> has been described and illustrated as including the fluid separation barrier <NUM> for separating the equilibration solution <NUM> from the vitrification solution <NUM>, in some embodiments, a specimen container may include a solid solution separation barrier that can undergo a solid to liquid phase change within a lumen of the specimen container. For example, <FIG> illustrate a specimen container <NUM> that includes such a separation barrier <NUM>.

The specimen container <NUM> is otherwise substantially similar in construction and function to the specimen container <NUM>. For example, as discussed above with respect to the specimen container <NUM>, the specimen container <NUM> can be used to prepare a specimen <NUM> according to a biological or other protocol and to subsequently store the specimen <NUM> in a low temperature substance in a viable and vitrified state. Furthermore, the specimen container <NUM> is a sterile, single-use device that is non-toxic to specimens contained therein. The specimen container <NUM> may be packaged as a single unit, and both the specimen container <NUM> and the packaging will remain sterile for a guaranteed shelf-life of the specimen container <NUM>. A total length of the specimen container <NUM> typically allows the specimen container <NUM> to fit within standard storage containers and other standard equipment used in ART protocols.

The specimen container <NUM> includes the elongate tube <NUM>, the ID label <NUM> (refer to <FIG>), the proximal closure <NUM>, the distal closure <NUM>, the equilibration solution <NUM>, and the vitrification solution <NUM> of the specimen container <NUM>. The separation barrier <NUM> between the solutions <NUM>, <NUM> is an inert substance in a solid phase below a threshold temperature and undergoes a solid to liquid phase change when heated to at least the threshold temperature. In some embodiments, the threshold temperature is about <NUM>. The specimen container <NUM> is stored at a refrigeration temperature of about <NUM> such that separation barrier is in a solid state. When warmed to a room temperature of about <NUM>, the separation barrier <NUM> is in a liquid phase. The solutions <NUM>, <NUM> are immiscible with the separation barrier <NUM> in a liquid phase such that the solutions <NUM>, <NUM> and the separation barrier <NUM> will not mix to form a homogeneous solution.

Example substances of the separation barrier <NUM> include alkanes, such as hexadecane (e.g., mineral oil or paraffin oil) and other alkanes (e.g., tetradecane through isocane) that melt from a solid to a liquid in a temperature range of about <NUM> (e.g., freezing) to about <NUM> (e.g., body temperature). In some embodiments, the separation barrier <NUM> may alternatively be provided by other, non-straight chain polymers, such as decanoic acid and caprylic acid. <FIG> illustrates the separation barrier <NUM> in a solid phase, and <FIG> illustrates the separation barrier <NUM> in a liquid phase following heating of the specimen container <NUM> to or above the threshold temperature (e.g., about <NUM> for hexadecane). A density of the separation barrier <NUM> in the liquid phase (e.g., about <NUM>/mL to about <NUM>/mL for hexadecane) is less than that of both solutions <NUM>, <NUM>.

In some cases, including a gaseous solution barrier (e.g., air) within a specimen container introduces a possibility of undesired gas bubble formation within the specimen container. Providing the separation barrier <NUM> as a solid substance (e.g., as opposed to a gaseous or liquid substance) within the specimen container <NUM> advantageously enables packaging of the solutions <NUM>, <NUM> in a separated state in a gas-free environment. Packaging the solutions <NUM>, <NUM> in a gas-free environment can prevent the formation of air bubbles within the specimen container <NUM>. Such air bubbles may otherwise cause problems with processing the specimen <NUM> within the specimen container <NUM>, such as obscuring visualization of the specimen <NUM>.

The specimen container <NUM> can be used to process the specimen <NUM> in the system console <NUM> as substantially described above with respect to the specimen <NUM> within the specimen container <NUM>. During processing of the specimen <NUM>, with the separation barrier <NUM> in a liquid state, g-forces exerted on the specimen container <NUM> can cause the solutions <NUM>, <NUM> and the separation barrier <NUM> to move past each other to desired positions along the elongate tube <NUM> based on differences in density between the solutions <NUM>, <NUM> and the separation barrier <NUM>. Movement (e.g., floatation) of the separation barrier <NUM> away from its initial position in which the separation barrier <NUM> continuously wets an inner surface of the elongate tube <NUM> (e.g., substantially about an entire circumference of the inner surface) opens up the lumen to permit movement of the solutions <NUM>, <NUM> and the specimen <NUM> distally within the elongate tube <NUM> according to the specimen processing protocol carried out by the system console <NUM>. Such behavior will be explained in more detail below with respect to a specimen container <NUM>.

Referring to <FIG>, the specimen container <NUM> is similar in function to the specimen container <NUM> and accordingly can be used to prepare a specimen <NUM> according to a biological or other protocol and to subsequently store the specimen <NUM> in a low temperature substance in a viable and vitrified state. Furthermore, the specimen container <NUM> is a sterile, single-use device that is non-toxic to specimens contained therein. The specimen container <NUM> may be packaged as a single unit, and both the specimen container <NUM> and the packaging will remain sterile for a guaranteed shelf-life of the specimen container <NUM>. A total length of the specimen container <NUM> typically allows the specimen container <NUM> to fit within standard storage containers and other standard equipment used in ART protocols. The specimen container <NUM> includes an elongate tube <NUM>, the identification label <NUM> (refer to <FIG>), a proximal closure that hermetically seals a proximal end of the elongate tube <NUM>, and a distal closure <NUM> that hermetically seals a distal end <NUM> of the elongate tube <NUM>, as discussed above with respect to like components of the specimen container <NUM>.

The elongate tube <NUM> is a thin capillary tube that has a very small variable diameter that changes in a stepwise manner along an axis <NUM> of the elongate tube <NUM>. The elongate tube <NUM> is internally preloaded with a culture media <NUM> in a main portion <NUM>, the equilibration solution <NUM> in an intermediate portion <NUM>, and the vitrification solution <NUM> in a distal portion <NUM>. Separation barriers <NUM>, <NUM> separate the culture media <NUM> and the solutions <NUM>, <NUM> from each other. The main portion <NUM> and the intermediate portion <NUM> respectively define beveled wall sections <NUM>, <NUM> that provide a transitional diameter between adjacent portions of the specimen container <NUM> for facilitating distal fluid flow in the specimen container <NUM> within a system console. In a temperature range of about <NUM> to about <NUM>, the culture media <NUM>, the solutions <NUM>, <NUM>, and the separation barriers <NUM>, <NUM> are all in a liquid phase. The culture media <NUM> contains various nutrients and molecules in concentrations that maintain viability of the specimen <NUM>.

The elongate tube <NUM> is dimensioned and a volume of the separation barrier <NUM> is selected such that the separation barrier <NUM> can continuously wet an internal surface of the intermediate portion <NUM>, but cannot continuously wet an internal surface of the main portion <NUM>. Similarly, the separation barrier <NUM> can continuously wet an internal surface of the distal portion <NUM>, but cannot continuously wet the internal surfaces of the intermediate portion <NUM> or the main portion <NUM>. Substance formulations of the separation barriers <NUM>, <NUM> are the same as those of the separation barrier <NUM>. Accordingly, the separation barriers <NUM>, <NUM> are inert substances in a solid phase below the threshold temperature and undergo a solid to liquid phase change when heated to at least the threshold temperature.

The volume of the equilibration solution <NUM> within the elongate tube <NUM> is typically about <NUM>µL to about <NUM>µL, and the volume of the vitrification solution <NUM> within the elongate tube <NUM> is typically about <NUM>µL to about <NUM>µL. The volume of the culture media <NUM> within the elongate tube <NUM> is typically about <NUM>µL to about <NUM>µL. Within the specimen container <NUM>, a density of the specimen <NUM> is greater than the densities of the solutions <NUM>, <NUM>, which are greater than a density of the culture media <NUM>, which is greater than the density of the separation barriers <NUM>, <NUM> in a liquid phase.

The main portion <NUM> of the elongate tube <NUM> typically has a length of about <NUM> to about <NUM> (e.g., about <NUM>) and an internal diameter of about <NUM> to about <NUM> (e.g., about <NUM>). The intermediate portion <NUM> of the elongate tube <NUM> typically has a length of about <NUM> to about <NUM> (e.g., about <NUM>) and an internal diameter of about <NUM> to about <NUM> (e.g., about <NUM>). The distal portion <NUM> of the elongate tube <NUM> typically has a length of about <NUM> to about <NUM> (e.g., about <NUM>) and an internal diameter of about <NUM> to about <NUM> (e.g., about <NUM>). The elongate tube <NUM> typically has a wall thickness of about <NUM> to about <NUM> (e.g., about <NUM>). The elongate tube <NUM> is manufactured via the techniques described above with respect to the elongate tube <NUM>. The elongate tube <NUM> is typically transparent or translucent and is typically made of the same materials as those from which the elongate tube <NUM> is made.

During a process for vitrifying the specimen <NUM> within the specimen container <NUM>, the proximal closure is removed from the elongate tube <NUM>, and a delivery device (e.g., such as the delivery device <NUM>) is used to deliver the specimen <NUM>, suspended within a small amount of culture media <NUM> to the culture media <NUM> within the main portion <NUM> of the elongate tube <NUM> through a proximal opening. The proximal closure is then reinstalled to the elongate tube <NUM> to reseal the proximal opening for further processing of the specimen <NUM>.

<FIG> illustrates a series of steps by which the specimen <NUM> is processed within the specimen container <NUM>. Step <NUM> illustrates an initial state of the specimen container <NUM> upon delivery of the specimen <NUM> to the culture media <NUM>, in which state the specimen container <NUM> has been pre-warmed to at least room temperature such that the barriers <NUM> and <NUM> are in a liquid phase. The specimen container <NUM> is subsequently loaded in a rotatable receptacle within a system console for exposure to applied g-forces. Step <NUM> illustrates a next state of the specimen container <NUM> following exposure to the applied g-forces within the system console. The g-forces urge the relatively dense specimen <NUM> and fluids (e.g., the culture media <NUM> and the solutions <NUM>, <NUM>) distally (e.g., to the right in <FIG>) into adjacent portions of the specimen container <NUM> and urges the relatively less dense fluid (e.g., the separation barriers <NUM>, <NUM>) proximally (e.g., to the left in <FIG>). Accordingly, the liquid separation barriers <NUM>, <NUM> separate from the internal surfaces of the intermediate and distal portions <NUM>, <NUM> and migrate (e.g., float) proximally as droplets within the specimen container <NUM>.

Step <NUM> of <FIG> illustrates that with continued exposure to the applied g-forces, the fluids within the specimen container <NUM> have arranged themselves in order of density, with the least dense fluid (e.g., the liquid separation barriers <NUM>, <NUM>) being positioned at a proximal region of the specimen container <NUM> and the most dense fluid (e.g., the vitrification solution <NUM>) remaining at the distal closure <NUM> of the specimen container. Step <NUM> illustrates that with further exposure to the applied g-forces, the specimen <NUM> (e.g., the most dense substance within the specimen container <NUM>) has been driven into the vitrification solution <NUM> near the distal closure <NUM>. The specimen container <NUM>, with the specimen <NUM> contained therein, can be subsequently submerged in a low temperature substance for vitrification and storage of the specimen <NUM>.

In some embodiments, a proximal closure of a specimen container can be used to deliver a specimen to the specimen container. For example, referring to <FIG>, a specimen container <NUM> includes such a proximal closure <NUM>. The specimen container <NUM> is otherwise substantially similar in construction and function to the specimen container <NUM> described above. For example, the specimen container <NUM> further includes the elongate tube <NUM>. The proximal closure <NUM> of the specimen container <NUM> includes the plug <NUM> and the top flange <NUM> of the specimen container <NUM>, as well as the specimen carrier <NUM> of the specimen container <NUM>.

<FIG> illustrates a series of steps by which a specimen <NUM> is loaded into the elongate tube <NUM> using the proximal closure <NUM>. In a first step, the delivery device <NUM> is used to aspirate (e.g., draw up) the specimen <NUM> and a small amount of culture media <NUM> from a petri dish <NUM>. In a second step, the specimen <NUM> and the culture media <NUM> are delivered to the specimen carrier <NUM> of the proximal closure <NUM>. In a subsequent step, the proximal closure <NUM>, carrying the specimen <NUM> atop the specimen carrier <NUM>, is inserted within the elongate tube <NUM> to close the elongate tube <NUM>. The specimen container <NUM>, carrying the specimen <NUM>, is subsequently placed within a system console <NUM> for processing the specimen <NUM> within the specimen container, as substantially described above with respect to processing of the specimen <NUM> within the specimen container <NUM>.

Other embodiments of system consoles for processing a specimen within a specimen container are also possible. For example, <FIG> illustrates such a system console <NUM> for processing a specimen within a specimen container <NUM> that may represent any of the above-discussed specimen containers. The system console <NUM> includes multiple processing stations <NUM> for supporting respective specimen containers <NUM> to carry out a specimen processing protocol, a platform <NUM> to which the processing stations <NUM> are secured, a housing <NUM> that supports that platform <NUM>, handles <NUM> for lifting or otherwise moving the system console <NUM>, and a lid <NUM> that is openable to allow access to the processing stations <NUM>. The system console <NUM> further includes a user interface screen <NUM>, multiple selectors <NUM> (e.g., buttons) for setting various parameters of the system console <NUM>, and a power switch <NUM> that are positioned along a front side of the housing <NUM>. The housing <NUM> is configured to rest atop a floor or another flat surface. The lid <NUM> is movable manually (e.g., pivotable, slidable, or removable) with respect to the housing <NUM>.

Additionally, the system console <NUM> includes a timer <NUM> (illustrated schematically), a reader component <NUM> (e.g., an RFID antenna or another type of reader component, illustrated schematically) that is programmed to read the ID label <NUM> of a specimen container, and a control module <NUM> (e.g., a microcontroller, illustrated schematically) that is programmed to control various features and functionalities of the system console <NUM>. The reader component <NUM>, the timer <NUM>, and the control module <NUM> may be positioned at respective locations within the system console <NUM> that are suitable for their respective functions. The user interface screen <NUM> allows a user to input several parameters that govern operation of the system console <NUM> to vitrify the specimen, such as a stage of the specimen (e.g., an oocyte or a blastocyst protocol selection). The user interface screen <NUM> may be an integrated touchscreen or a touchless screen associated with tactile control elements, such as buttons, knobs, dials, or the like. The control module <NUM> includes one or more processors that are in communication with and/or are programmed to control various actuators and sensors of the system console <NUM> related to various automated features, such as receiving and instantiating user selections input at the user interface screen <NUM>, reading the ID label <NUM> of the specimen container <NUM>, executing the timer <NUM>, spinning the platform <NUM> at a specified spin speed for a specified duration, detecting an open/closed state of the lid <NUM>, and providing audible and/or visual feedback regarding a progression of the process.

The platform <NUM> defines multiple tracks <NUM> at which a processing station <NUM> can be secured in a fixed in position with respect to the platform <NUM>. Each processing station <NUM> includes a lower support bracket <NUM> and an upper support bracket <NUM> that together define a receptacle <NUM> for holding the specimen container <NUM>. Each processing station <NUM> further includes an imaging system <NUM> by which movement of the specimen within the specimen container <NUM> can be observed. The reader component <NUM> can detect a presence of the specimen container <NUM> within the receptacle <NUM> by reading the ID label <NUM> (e.g., the RFID tag) and can communicate such detection to the control module <NUM>, which can cause the timer <NUM> to be activated.

According to one or more signals received from the control module <NUM>, the platform <NUM> can spin about a central axis <NUM> to exert enough centripetal force on the specimen to cause the specimen to move along an axis <NUM> of the specimen container <NUM> toward a distal end <NUM> according to a specified protocol. During spinning of the platform <NUM>, the specimen and various processing media (e.g., equilibration and vitrification solutions and other media) within the specimen container <NUM> can be visualized by the imaging system <NUM>. The control module <NUM> can adjust an angular speed of the platform <NUM> and/or a duration of one or more phases of the protocol based on feedback from the imaging system <NUM> regarding a position of the specimen. Such protocol adjustments can optimize time periods of specimen exposure to the processing media within the specimen container <NUM>. Upon completion of the processing protocol, the specimen container <NUM> is removed from the processing station <NUM> and placed within a low temperature substance for vitrification and cryopreservation of the specimen contained within the specimen container <NUM>.

In some embodiments, the system console <NUM> has a total length of about <NUM> to about <NUM>, a total width of about <NUM> to about <NUM>, and a total height of about <NUM> to about <NUM>. In some embodiments, the system console <NUM> has a weight in a range of about <NUM> to about <NUM> and is typically stored on a laboratory floor, a storage facility floor, a table, or a countertop, that has an ambient environmental temperature of about <NUM> to about <NUM>. In some embodiments, the receptacle <NUM> of the processing station <NUM> has a length of about <NUM> to about <NUM> and a width of about <NUM> to about <NUM>. The support brackets <NUM>, <NUM> of the processing station <NUM> and the platform <NUM> are typically made of metal. The housing <NUM> and the lid <NUM> are typically made of materials that provide a degree of thermal insulation, such as polymers.

Claim 1:
A specimen container (<NUM>; <NUM>; <NUM>; <NUM>; <NUM>; <NUM>; <NUM>; <NUM>; <NUM>; <NUM>; <NUM>; <NUM>; <NUM>; <NUM>; <NUM>; <NUM>; <NUM>; <NUM>; <NUM>; <NUM>) configured for cryogenic processing of a specimen, the specimen container comprising:
an elongate member (<NUM>; <NUM>; <NUM>; <NUM>; <NUM>; <NUM>; <NUM>; <NUM>; <NUM>);
an equilibration solution (<NUM>; <NUM>) preloaded within a lumen of the elongate member at a first position;
a vitrification solution (<NUM>; <NUM>, <NUM>, <NUM>) preloaded within the lumen of the elongate member at a second position located distal to the first position; and
a displaceable barrier (<NUM>; <NUM>; <NUM>; <NUM>; <NUM>; <NUM>, <NUM>) positioned between the equilibration solution and the vitrification solution such that the equilibration solution and the vitrification solution are initially spaced apart from each other within the lumen of the elongate member.