Cellular cryobiology: thermodynamic and mechanical effects

Several physical stresses kill cells at low temperatures. Intracellular ice is usually fatal, so survival of freezing temperatures involves combinations of dehydration, freezing point depression, supercooling and intracellular vitrification. Artificial cryopreservation achieves intracellular vitrification with rapid cooling, modest osmotic contraction and, often, added cryoprotectants. High warming rates are required to avoid crystallization during warming. Environmental cooling is much slower and temperatures less cold, but environmental freezing damage is important ecologically and agronomically. For modest sub-freezing temperatures, supercooling sometimes allows survival. At lower temperatures, extracellular water usually freezes and cells may suffer large osmotic contractions. This contraction concentrates solutes and thus assists vitrification, but is not necessarily reversible: the rapid osmotic expansion during thawing may rupture membranes. Further, membranes and other ultrastructural elements may be damaged by the large, anisotropic mechanical stresses produced when their surfaces interact via hydration forces. Solutes reduce these stresses by osmotic, volumetric and other effects.     

Ice storage system with water–oil mixture: formation of suspension with high IPF

A new method of forming ice, which is one of the dynamic types of ice storage system, is studied. In the method a water-oil emulsion is cooled with stirring in a vessel and changed into an ice-oil and water suspension. A mixture of 10 vol% silicone-oil and 90 vol% water is emulsified with a small amount of an additive. Silane-couplers are tested as the additive and effects of the additive on ice formation process are investigated. Cooling rate is changed and vessels made of various materials are tested. It is proved that the present method has the following characteristics. Ice–oil and water suspension (slush ice) which has a good fluidity is able to be formed without adhering to the cooling surface. Ice in the suspension is granular and dispersed state and the suspension with more than 70% of ice packing factor (IPF) is also able to be formed. The suspension with the high IPF can be preserved for a long time in the granular state.     

Three-dimensional measurement of ice crystals in frozen beef with a micro-slicer image processing system

A novel technique has been developed for measuring the three-dimensional (3-D) structure and distribution of ice crystals formed in frozen beef by using a micro-slicer image processing system (MSIPS). The system has functions to reconstruct the 3-D image based on the image data of exposed cross-sections obtained by multi-slicing of a frozen sample with the minimum thickness of 1 μm and to display the internal structure as well as an arbitrary cross-section of the sample choosing observation angles. The size and distribution of ice crystals can be determined from the 2-D quantitative information, such as the periphery and area of the crystals. The effects of freezing conditions on the morphology and distribution of the ice crystals were demonstrated quantitatively from the observations of raw beef stained by fluorescent indicator. For the samples frozen at −15 °C, the network structure of ice crystals were observed mainly at intercellular space, having approximately 100 μm in cross-sectional size, while that prepared at −120 °C showed the spherical crystals of 10–20 μm in diameter within the cells. The 3-D image of the sample demonstrated that the growth of ice columns was restricted by the intrinsic structure of muscle fibers. The proposed method provided a new tool to investigate the effects of freezing conditions on the size, morphology and distribution of ice crystals.     

Ice storage system using water–oil mixture Discussion about influence of additive on ice formation process

Ice storage is one technique for effective use of thermal energy. So, many studies on slush ice as a thermal storage material have been done. We have also been studying a suspension (slush ice) made from an oil-water mixture by cooling and stirring. From our study results, it was found that an additive having both an amino group (-NH₂) and a silanol group (-SiOH) was essential to form a suspension with high IPF without adhesion of ice to the cooling wall. Moreover, ice particles formed in the suspension were dispersed and granular, and did not stick to each other. In the present paper, we carried out experiments to clarify the characteristics of the suspension formation process. From a thermal analysis of the substance formed in the suspension by difference scanning calorimeter (DSC), it was found that the substance was not ice but a compound of ice and additive. Then, at a very small depression of freezing point (about 7°C) all water in the mixture could be frozen by using the additive.     

Study on prevention of ice adhesion to cooling wall due to voltage impression in ice storage—discussion on possibility of attraction of oil to wall

For ice storage, one of authors has studied new ice slurry formed by cooling a water–oil mixture with stirring. When the mixture is stirred in a vessel, oil is charged by static electricity due to friction. If the vessel wall can attract charged oil, prevention of ice adhesion to the wall may be realized. Therefore, in this paper, in order to observe behavior of charged water–oil droplet or mixture in electric field by a high speed camera or video camera, two types of experiments were carried out. One was that the water–oil droplet charged by static electricity was made to fall plumb down between two electrodes with electric field or without electric field, varying the water content of droplet. The other was that a constant voltage was applied on the vessel filled with the water–oil mixture stirred. From experiments, it was confirmed that attracting force between the charged wall (electrode) and charged oil acted.     

Continuous ice slurry formation by using W/O emulsion with higher water content

A W/O (water-in-oil) emulsion was made from a water–lamp oil mixture with higher water content and a small amount of an additive of amino group-modified silicone oil, and the emulsion could be changed into an ice slurry by cooling with stirring. By using a new continuous ice formation system proposed by one of the authors of this paper, the ice slurry could be formed continuously and stably in an ice formation vessel made of stainless steel. From the experimental results, the conditions were clarified for realizing continuous ice formation for 10 h without ice adhesion to the cooling wall. Moreover, in order to propagate supercooling dissolution of the emulsion effectively and to decrease viscosity in the ice slurry, voltages were applied to the emulsion and ice slurry formed, respectively, and it was clarified that the voltage impression was effective for both.     

Study on continuous ice slurry formation using functional fluid for ice storage – Discussion of optimal operating conditions

A functional fluid was made by adding a small amount of additive to a water–silicone-oil mixture with 90 vol% water content, and the functional fluid was transformed into an ice slurry by cooling while stirring. The new ice formation system, which authors proposed for ice storage based on the results of previous studies, demonstrated that the ice slurry could be formed continuously for 10 h. In the current paper, experiments were carried out, varying operating conditions, and an optimal operating condition was determined to improve performance of the present system still more. From the experimental results, the conditions necessary to increase the amount of recovery ice were clarified. The time-dependencies of the shape and size of formed ice particles were also shown. Moreover, the reason why the freezing temperature of the functional fluid rose due to repetition of ice formation was clarified, and its measure was discussed. The present study then found that it was possible to form and recover a larger amount of ice than in previous attempts, given the rise in freezing temperature.     

Continuous ice slurry formation using a functional fluid for ice storage

A functional fluid composed of an oil–water mixture with an additive is transformed into an ice slurry by cooling while stirring. This paper describes a new continuous ice slurry formation method. Experiments were carried out by varying conditions such as the supply time of functional fluid, the stirrer torque, brine temperature and degree of supercooling. As a result, the characteristics of the ice formation and recovery processes were clarified. It was found that the ice particles gradually became uniform in size and spherical, and grew to 3.5 mm in diameter during about 10 h. The factors influencing the size of formed ice particles were discussed because the larger ice particles were expected to melt more rapidly. The ice particle size was found to increase with decreasing degree of supercooling and cooling rate, and with increasing stirrer wing diameter.     

Application of thermal battery in the ice storage air-conditioning system as a subcooler

This article experimentally investigates the thermal performance of a thermal battery used in the ice storage air-conditioning system as a subcooler. The thermal battery utilizes the superior heat transfer characteristics of two-phase closed thermosyphon and eliminates the drawbacks found in convectional energy storage systems. Experimental investigations are first conducted to study the thermal behavior of thermal battery under different charge temperatures (−5 °C to −9 °C) in which water is used as the energy storage material. This study also examines the thermal performance of the subcooled ice storage air conditioner under different cooling loads. Experimental data of temperature variation of water, ice fraction, refrigerant mass flow rate and coefficient of performance (COP) are obtained. The results show that supercooling phenomenon appears in the water and it can be ended when the charge temperature is lower than −6 °C. The system gives 28% more cooling capacity and 8% higher COP by the contribution of the thermal battery used as a subcooler.     

Formation of high performance ice slurry by W/O emulsion in ice storage (effective method to propagate supercooling dissolution)

This study focused ice slurry formation in an ice storage system using W/O emulsions with 70 and 80% water contents. Emulsions consisted of a silicone oil–water mixture with a small amount of amino-group-modified silicone oil additive. Ice slurry was formed by cooling the emulsion without ice adhesion to the cooling wall, as water in the emulsion did not directly contact the cooling wall. As the structure of W/O emulsion slowed the propagation rate of supercooling dissolution, voltage and ultrasonic wave were applied to the W/O emulsion to propagate dissolution more quickly and decrease maximum supercooling degree, respectively. Thus, the effects of voltage and ultrasonic wave applications on propagation rate were clarified.     

Study on high performance ice slurry formed by cooling emulsion in ice storage (discussion on adaptability of emulsion to thermal storage material)

This study focuses on an emulsion as a new thermal storage material for ice storage. Two types of emulsions were formed using an oil–water mixture with a small amount of additive. A silicone, light and lump oils were used. The water contents of the emulsions were 70, 80 and 90%. The additive was an amino group modified silicone oil. No depression of freezing point was observed for the emulsions because of their hydrophobic properties. In order to determine the structure of the emulsions, their electrical resistances were measured. Moreover, components of the liquids separating from the emulsions were analyzed. The results indicated that one emulsion was a W/O type emulsion, while the other was an O/W type. Finally, adaptability of the two emulsions to ice storage was discussed, it was concluded that a high performance ice slurry could be formed by the W/O type emulsion.     

Cryobank of plant genetic resources in Russian Academy of Sciences

Cryopreservation is the most reliable method for long-term storage of plant genetic resources. A review of cell injury by ice crystals and dehydration during a freeze–thaw cycle is given. For successful regeneration of plants and cultures after cryopreservation of their cells, the development of reliable cryopreservation procedure is required including preliminary cultivation, treatment by cryoprotectors, freezing by different methods, thawing and recultivation. Up to now 27 cell lines successfully resumed their growth after storage in liquid nitrogen and preserved their specific features and biosynthetic potential. Besides, shoot tips of 40 cultivars of potato, rose, strawberry and raspberry regenerated plants both in vitro and in vivo after cryopreservation. The longest storage duration was 25 years. Now in liquid nitrogen we continuously store 24 cell strains of rare medicinal plants, shoot tips of seven cultivars of strawberry and raspberry and seeds of 250 endangered plant species collected over all Russian territory.     

Evaluation of the mean ice ratio as a function of temperature in a heterogeneous food: Application to the determination of the target temperature at the end of freezing

The freezing process is widely used in the food industry. In the 70s, French regulation authorities have created in collaboration with the food industry the concept of «surgélation» process with the objective of improving the image of high quality frozen foods. The process of “surgélation” which could be translated as “super freezing” corresponds to a freezing process for which a final temperature of −18 °C must be reached “as fast as possible”. This concept was proposed in opposition to a conventionally “freezing” process for which no specific freezing rate is expected and the final storage temperature can be of −12 °C only. The objective of this work is to propose a methodology to evaluate the mean amount of frozen ice in a complex food as a function of temperature and to deduce a target temperature that must be considered as the temperature for which the food may be considered as “frozen”. Based on the definition proposed by the IIF-IIR red book, this target temperature has been defined as the temperature for which 80% of the freezable water is frozen. A case study is proposed with a model food made of two constituents.     

Measurement of thermal conductivity of ice slurry made from solution by transient line heat-source technique (analytical discussion on influence of latent heat of fusion)

We have been studying on ice slurry in a dynamic type ice storage system. The ice slurry has many good characteristics. The ice slurry can be made from a solution. When designing the ice storage system using this ice slurry, thermal conductivity of the ice slurry is essential.When thermal conductivity of the ice slurry made from a solution is measured by a transient line heat-source technique, a measurement value of thermal conductivity is affected by a latent heat of fusion of ice. Therefore, the thermal conductivity measured is apparent thermal conductivity. In this study, influences of Stefan number, initial concentration of the solution, initial solid fraction (initial IPF) and Fourier number on the thermal conductivity was analytically discussed to improve measurement accuracy of the thermal conductivity of ice slurry in the transient line heat-source technique.     

Development of computer program for simulation of an ice bank system operation, Part I: Mathematical modelling

Since the use of standard engineering methods in the process of an ice bank performance evaluation offers neither adequate flexibility nor accuracy, the aim of this research was to provide a powerful tool for an industrial design of an ice storage system allowing to account for the various design parameters and system arrangements over a wide range of time varying operating conditions. In this paper the development of a computer application for the prediction of an ice bank system operation is presented. Static, indirect, cool thermal storage systems with external ice on coil building/melting were considered. The mathematical model was developed by means of energy and mass balance relations for each component of the system and is basically divided into two parts, the model of an ice storage system and the model of a refrigeration unit. Heat transfer processes in an ice silo were modelled by use of empirical correlations while the performance of refrigeration unit components were based on manufacturers data. Programming and application design were made in Fortran 95 language standard. Input of data is enabled through drop down menus and dialog boxes, while the results are presented via figures, diagrams and data (ASCII) files. In addition, to demonstrate the necessity for development of simulation program a case study was performed. Simulation results clearly indicate that no simple engineering methods or rule of thumb principles could be utilised in order to validate performance of an ice bank system properly.     

Investigation into the transportation and melting of thick ice slurries in pipes

This paper presents the results of experiments and modelling carried out on ice slurries flowing in uninsulated steel pipes with a nominal diameter of 50 mm. The slurries used were formed from 4.75% NaCl aqueous solution and had ice mass fractions in the range 18–42%, with a view to the use of thick ice slurry ‘pigs’ as a pipeline clearing technique. Of particular interest was the distance over which such slurries can survive as plug-like entities, before melting reduces them to ineffective thin two-phase suspensions. The experiments showed that for small volumes of slurry, survivability is directly proportional to the quantity of slurry used, but that increasing the ice fraction has a more marked effect. A simple one-dimensional numerical model that accounts for transportation, heat transfer and melting was developed that produces reasonable predictions.     

Ice-water two-phase flow behavior in ice heat storage systems

In this paper, we describe (1) the characteristics of ice-water two-phase flow created in a supercooling-type ice storage system, (2) the numerical analysis models for both processes of ice storing and melting within ice storage tanks, and (3) the design technologies for ice storage systems based on such predictive models, as well as the operational performance of actual equipment built with these design technologies. As the result of this technological development, it has been confirmed that the predictive approaches for the one dimensional ice accumulation in the given tanks and ice melting with low temperature water enough to air conditioning applications are reasonable and widely usable. Furthermore by the design conditions obtained here it is possible to simplify the construction of ice storage tanks and to realize high reliability in the actual equipment.     

Experimental investigation of ice slurry heat transfer in horizontal tube

Heat transfer of ice slurry flow based on ethanol–water mixture in a circular horizontal tube has been experimentally investigated. The secondary fluid was prepared by mixing ethanol and water to obtain initial alcohol concentration of 10.3% (initial freezing temperature -4.4 °C). The heat transfer tests were conducted to cover laminar and slightly turbulent flow with ice mass fraction varying from 0% to 22% depending on test performed. Measured heat transfer coefficients of ice slurry are found to be higher than those for single phase fluid, especially for laminar flow conditions and high ice mass fractions where the heat transfer is increased with a factor 2 in comparison to the single phase flow. In addition, experimentally determined heat transfer coefficients of ice slurry flow were compared to the analytical results, based on the correlation by Sieder and Tate for laminar single phase regime, by Dittus–Boelter for turbulent single phase regime and empirical correlation by Christensen and Kauffeld derived for laminar/turbulent ice slurry flow in circular horizontal tubes. It was found that the classical correlation proposed by Sieder and Tate for laminar forced convection in smooth straight circular ducts cannot be used for heat transfer prediction of ice slurry flow since it strongly underestimates measured values, while, for the turbulent flow regime the simple Dittus–Boelter relation predicts the heat transfer coefficient of ice slurry flow with high accuracy but only up to an ice mass fraction of 10% and Re_cf > 2300 regardless of imposed heat flux. For higher ice mass fractions and regardless of the flow regime, the correlation proposed by Christensen and Kauffeld gives good agreement with experimental results.     

Extension of soil thermal conductivity models to frozen meats with low and high fat content

Thermal conductivity models of frozen soils were analyzed and compared with similar models developed for frozen foods. In total, eight thermal conductivity models and 54 model versions were tested against experimental data of 13 meat products in the temperature range from 0 to −40 °C. The model by deVries, with water+ice (wi) as the continuous phase, showed overall the best predictions. The use of wi leads generally to improved predictions in comparison to ice; water as the continuous phase is beneficial only to deVries model, mostly from −1 to −20 °C; fat is advantageous only to meats with high fat content. The results of this work suggest that the more sophisticated way of estimating the thermal conductivity for a disperse phase in the deVries model might be more appropriate than the use of basic multi-phase models (geometric mean, parallel, and series). Overall, relatively small differences in predictions were observed between the best model versions by deVries, Levy, Mascheroni, Maxwell or Gori as applied to frozen meats with low content of fat. These differences could also be generated by uncertainty in meat composition, temperature dependence of thermal conductivity of ice, measurement errors, and limitation of predictive models.     

Freezing of a water droplet due to evaporation—heat transfer dominating the evaporation–freezing phenomena and the effect of boiling on freezing characteristics

One of the authors has proposed a novel transport/storage system for the waste cold from the gasification process of liquefied natural gas (LNG), which consists of an evaporator, a cold trap, and a pipeline. In order to estimate the performance of this system, one should know the pressure in the evaporator, in which evaporation–freezing of a PCM occurs, and in the cold trap, as well as the pressure drop of the pipeline due to the flow of low pressure vapor of the PCM. In this paper, the cooling/freezing phenomena of a water droplet due to evaporation in an evacuated chamber was experimentally examined, and the heat transfer dominating the evaporation-freezing phenomena was investigated in order to estimate the pressure in the evaporator. From the results, it was shown that the water droplet in the evacuated cell is effectively cooled by the evaporation of water itself, and is frozen within a few seconds through a remarkable supercooling state, and that the cooling rate of the water droplets were dominated by heat transfer within the droplet under the abrupt evacuation condition. The later result means that, in order to obtain an ice particle by evaporation–freezing, the surroundings of the water droplet should be evacuated at the pressure as low as the saturate pressure of water at the maximum supercooling temperature of the droplet.