Method and apparatus for cryopreserving biological specimens
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Solution Overview
Problem
Current snap freezing techniques for cryopreservation lack standardization and control over cooling rates, leading to inconsistent sample preservation and potential damage to biological materials during the freezing process.
Innovation Solution
A method and apparatus that involve pre-setting a cooling fluid to a first temperature between -10 °C and -60 °C for rapid freezing, followed by a controlled temperature reduction to a second predetermined temperature using a PID algorithm and cooling profile, ensuring a consistent cooling rate of 0.5 °C to 3 °C per minute, typically 1 °C per minute, to minimize ice crystal formation and osmotic imbalances.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Speed
If rapid snap freezing is performed using conventional methods, then freezing speed is improved, but cooling rate control and sample preservation consistency deteriorate
Solution Approach 1:
The cooling fluid is pre-cooled to a specific temperature range (-10°C to -60°C) before the specimen is introduced. This preliminary preparation ensures that when rapid freezing occurs, the cooling rate is optimized from the start, combining high freezing speed with controlled cooling parameters to prevent ice crystal formation while maintaining preservation consistency.
2Loss of time
If uncontrolled rapid cooling is applied, then freezing time is reduced, but sample integrity and cell viability deteriorate
Solution Approach 1:
The invention optimizes the cooling fluid temperature parameter to a specific range (-10°C to -60°C) rather than using extremely low temperatures. This parameter change enables rapid freezing that completes in minutes while maintaining sample integrity, avoiding the formation of large ice crystals and excessive osmotic stress that would damage cells and tissues.
3Manufacturing precision
If standardized cooling protocols are implemented, then preservation consistency is improved, but flexibility in handling different sample types deteriorates
Solution Approach 1:
The invention provides a standardized cooling protocol using a controlled cooling rate (0.5°C to 3°C per minute) that can be universally applied to various biological specimens including cells, tissues, and organoids. This standardized approach ensures preservation consistency across different sample types while the modular apparatus design maintains flexibility for handling diverse specimens.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach standardizes the cryopreservation process, reducing variables that affect cell viability and maintaining sample integrity by optimizing the cooling rate, thus ensuring maximum preservation quality for various biological specimens.
Implementation Method 1
a controlled temperature reduction to a second predetermined temperature using a PID algorithm and cooling profile, ensuring a consistent cooling rate of 0.5 °C to 3 °C per minute
Implementation Method 2
During cryopreservation, the water changes to ice and cellular metabolism ceases. Dehydration also occurs, changing the concentration of salts and other metabolites and creating an osmotic imbalance
Data Source
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AI summary
The present invention relates to a method for collecting and preserving biological specimens for biobanking cryopreservation, comprising the steps of i) placing the specimen in a cooling fluid pre-set at a first predetermined temperature of between -10°C and -60°C for snap freezing the specimen, and ii) reducing the temperature of said cooling fluid to a second predetermined temperature suitable for preserving said specimen. The second temperature is most preferably -80°C. The reduction of the temperature in step ii) is performed by a predetermined cooling profile. The invention is further directed to a corresponding apparatus (1), comprising a receiving means (2) for receiving a cooling fluid and the specimen, a cooling means to cool said fluid in said receiving means, and a controller configured to control the cooling means. The receiving means comprises at least one reservoir (3, 30) and a working plate (4).The cooling means is preferably a stirling cooler using Argon gas.