Method and apparatus for reducing probability of ice nucleation during preservation of biological matter in isochoric systems

A method for reducing probability of ice nucleation during preservation of biological matter in isochoric systems comprising:

• (a) placing biological matter in a flexible and impermeable inner container;
• (b) removing bulk gas from the inner container;
• (c) sealing the inner container;
• (d) placing the inner container in a rigid, non-thermally insulating outer container so as to create a space between an outer surface of the inner container and an inner surface of the outer container;
• (e) filling the space between the outer surface of the inner container and the inner surface of the outer container with an outer solution;
• wherein the biological matter has a melting point, and the outer solution has a melting point that is lower than the melting point of the biological matter;
• (f) removing bulk gas from the outer container;
• (g) sealing the outer container;
• (h) cooling the inner container, the outer container, the biological matter, and the outer solution to the desired storage temperature:
• (i) maintaining the inner container, the outer container, the biological matter, and the outer solution at the desired storage temperature for a desired storage period;
• (j) warming the inner container, the outer container, the biological matter, and the outer solution to a temperature that is higher than the desired storage temperature;
• (k) unsealing the outer container and the inner container;
• and
• (l) removing the biological matter from the inner container.

An apparatus for reducing probability of ice nucleation during preservation of biological matter in isochoric systems comprising:

• (a) an outer container that is rigid and non-thermally insulating;
• wherein the outer container comprises a seal that is configured to provide air- and liquid-tight sealing;
• (b) an inner container that is situated within the outer container;
• wherein the inner container is flexible but cannot transmit mass;
• (c) an inner solution within the inner container;
• wherein the inner solution has an equilibrium melting point that is above a desired sub-zero centigrade storage temperature;
• and
• (d) an outer solution within the outer container and outside of the inner container;
• wherein the outer solution is comprised of a liquid that has an equilibrium melting point that is below the desired sub-zero centigrade storage temperature.

An apparatus for reducing probability of ice nucleation during preservation of biological matter in isochoric systems comprising:

• (a) an outer container that is rigid and non-thermally insulating;
• wherein the outer container comprises a seal that is configured to provide air- and liquid-tight sealing;
• (b) an inner container that is situated within the outer container;
• wherein the inner container is flexible but cannot transmit mass;
• (c) an inner solution within the inner container;
• wherein the inner solution has an equilibrium melting point that is above a desired sub-zero centigrade storage temperature;
• and
• (d) an outer solution within the outer container and outside of the inner container;
• wherein the outer solution is configured to undergo vitrification at the desired sub-zero centigrade storage temperature.

An apparatus for reducing probability of ice nucleation during preservation of biological matter in isochoric systems comprising:

• (a) an outer container that is rigid and non-thermally insulating;
• wherein the outer container comprises a seal that is configured to provide air- and liquid-tight sealing;
• (b) at least two inner containers that are situated within the outer container;
• wherein the inner containers are flexible but cannot transmit mass;
• (c) an inner solution within each of the inner containers;
• wherein the inner solution has an equilibrium melting point that is above a desired sub-zero centigrade storage temperature;
• and
• (d) an outer solution within the outer container and outside of the inner containers;
• wherein the outer solution is comprised of a liquid that has an equilibrium melting point that is below the desired sub-zero centigrade storage temperature.

An apparatus for reducing probability of ice nucleation during preservation of biological matter in isochoric systems comprising:

• (a) an outer container that is rigid and non-thermally insulating;
• wherein the outer container comprises a seal that is configured to provide air- and liquid-tight sealing;
• (b) at least two inner containers that are situated within the outer container;
• wherein the inner containers are flexible but cannot transmit mass;
• (c) an inner solution within each of the inner containers;
• wherein the inner solution has an equilibrium melting point that is above a desired sub-zero centigrade storage temperature;
• and
• (d) an outer solution within the outer container and outside of the inner containers;
• wherein the outer solution is configured to undergo vitrification at the desired sub-zero centigrade storage temperature.