Method and device for determining catalytic efficiency of artificial ice nucleating material

Abstract

The method and device for determining the catalytic efficiency of the artificial ice nucleus material provided by the embodiments of the present disclosure comprise the following steps: determining the three-dimensional reconstruction graph and the lattice mismatch degree of the biological macromolecular multi-scale complex according to a preset first observation condition; determining the thermodynamic evaluation result based on the three-dimensional reconstruction graph, the lattice mismatch degree and a preset second observation condition; obtaining the process image of the newly generated ice crystal front surface based on a preset third observation condition; extracting the newly generated ice crystal front surface advancing contour line and the differentiated time track path from the process image according to a space reference; determining the catalytic rate order jump according to the advancing contour line and the differentiated time track path; obtaining the extreme catalytic efficiency quantitative result; realizing the analysis of the biological macromolecular multi-scale complex native hydration folding conformation; accurately positioning and quantifying the core nucleation active center; and providing the quantitative backstepping mechanism of the interface polarity and the thermodynamic nucleation potential barrier.

Description

Technical Field

[0001] This disclosure relates to the field of meteorological engineering technology, and in particular to a method and apparatus for determining the catalytic efficiency of an artificial ice core material. Background Technology

[0002] The core mechanism of freezing rain disasters lies in the instantaneous heterogeneous crystallization that occurs when supercooled droplets come into contact with the Earth's surface. Dispersing highly efficient artificial ice nuclei into clouds or near-surface layers to artificially induce premature crystallization of supercooled water in the air is currently an important physical means of artificially intervening in severe weather events such as freezing rain. However, heterogeneous nucleation is an instantaneous physical phase transition event spanning from the molecular to the micro-nano scale.

[0003] Limited by existing macroscopic observation and conventional microscopic techniques, especially for artificial ice nuclei made of highly efficient soft matter such as biological proteins, the microscopic catalytic mechanism of these nuclei has long remained a "black box," facing the following three major technical bottlenecks: First, conformational fidelity is difficult to maintain. Conventional freezing easily destroys the original hydrated folding conformation of fragile macromolecular ice nuclei, making it difficult to accurately resolve their microscopic topological structure as "ice-forming condensation templates." Second, thermodynamic orbit determination is difficult. Macroscopic temperature measurement equipment cannot accurately locate the "core nucleation active center" at the micro-nano scale, and there is a lack of quantitative reverse-engineering mechanisms for interfacial polarity and thermodynamic nucleation barriers. Third, kinetic quantification is difficult. There is a lack of advanced in-situ characterization methods for high-frequency capture under extreme non-equilibrium conditions, making it difficult to obtain the extremely rapid crystallization propulsion rate of the instantaneous transition from the gas / liquid phase to the solid phase. Summary of the Invention

[0004] In order to solve the above-mentioned technical problems, or at least partially solve the above-mentioned technical problems, this disclosure provides a method and apparatus for determining the catalytic efficiency of artificial ice core materials.

[0005] This disclosure provides a method for determining the catalytic efficacy of an artificial ice core material, the method comprising: The sample tilt angle and projection sequence of the biomacromolecule multiscale complex are obtained according to the preset first observation conditions. The sample tilt angle and projection sequence are reconstructed and decoded to obtain the three-dimensional reconstruction map and lattice mismatch of the biomacromolecule multiscale complex, wherein the biomacromolecule multiscale complex is embedded in an ice carrier. The observation field of view is determined based on the three-dimensional reconstruction map and the lattice mismatch. The nascent micro ice crystals are observed according to the observation field of view and the preset second observation conditions, and a nucleation spatial probability hotspot map is drawn. The nucleation spatial probability hotspot map is analyzed to obtain the thermodynamic evaluation result of the biomacromolecule multiscale complex. The nascent micro ice crystals are obtained by the biomacromolecule multiscale complex after a preset treatment. The process image of the formation of the new ice crystal front is obtained based on the preset third observation conditions. The advancement contour line of the new ice crystal front is extracted from the process image based on the spatial reference. The differentiated time trajectory path of the new ice crystal front in the preset region is determined according to the nucleation spatial probability hotspot map. The new ice crystal front is obtained by phase transition reaction of biomacromolecule multiscale complex. The spatial reference is obtained by the core nucleation active center identified in the thermodynamic evaluation results. A spatiotemporal map is obtained based on the differentiated time trajectory path and the propulsion contour line. The catalytic rate order-of-magnitude transition of the newly formed ice crystal front is determined based on the spatiotemporal map. Based on the catalytic rate order-of-magnitude transition, the quantitative result of the extreme catalytic efficiency induced by the multi-scale complex of the biomacromolecule to non-classical crystallization is obtained.

[0006] The method provided in this disclosure acquires the sample tilt angle and projection sequence of a multi-scale biomolecular complex according to preset first observation conditions, reconstructs and decodes the sample tilt angle and projection sequence to obtain a three-dimensional reconstruction image and lattice mismatch of the multi-scale biomolecular complex, wherein the multi-scale biomolecular complex is embedded in an ice carrier, comprising: Based on the environmental and equipment parameters in the preset first observation conditions, a series of tilt imaging was performed on the biomacromolecule multiscale complex using cryo-electron tomography to obtain the sample tilt angle and projection sequence of the biomacromolecule multiscale complex. The common structural features in each projection image of the series of tilt imaging are extracted. The sample tilt angle is aligned with the projection sequence based on the common structural features to obtain the alignment result. The alignment result is then reconstructed by back projection to obtain the three-dimensional electron density map of the biomacromolecule multiscale complex in the ice carrier. The three-dimensional electron density map is decoded to obtain the three-dimensional multipolymer conformation information of the biomacromolecule multiscale complex in the ice carrier, and a three-dimensional reconstruction map is drawn based on the three-dimensional multipolymer conformation information. Based on the three-dimensional reconstructed image, a confined region high-resolution imaging of the phase interface between the biomacromolecule multiscale complex and the ice carrier is performed, local fast Fourier transform is extracted, and the lattice mismatch of the biomacromolecule multiscale complex is calculated.

[0007] The method provided in this disclosure determines the observation field of view based on the three-dimensional reconstruction image and lattice mismatch, observes nascent micro-ice crystals according to the observation field of view and preset second observation conditions, and draws a nucleation spatial probability hotspot map. The method then analyzes the nucleation spatial probability hotspot map to obtain the thermodynamic evaluation result of the biomacromolecule multi-scale complex. The nascent micro-ice crystals are obtained by pre-processing the biomacromolecule multi-scale complex. The preset processing includes in-situ micro-sublimation and targeted re-sublimation, including: Based on the three-dimensional reconstruction image and lattice mismatch, the observation field of view is determined. Based on the observation field of view and the preset second observation conditions, the ice carrier containing the biomacromolecule multiscale complex is induced to perform in-situ micro-sublimation, exposing the characteristic surface and interface of the biomacromolecule multiscale complex. Based on the characteristic interface and the preset second observation conditions, targeted resublimation is performed on the biomacromolecule multiscale complex to obtain nascent micro ice crystals; Based on the three-dimensional reconstruction map, the nascent micro ice crystals are anchored in three-dimensional space coordinates, and a nucleation space probability hotspot map is drawn. The distribution of the nascent micro ice crystals is determined by analyzing the nucleation space probability hotspot map. Based on the distribution, high-probability aggregation regions of nascent micro ice crystals are extracted from the nucleation space probability hotspot map, and the high-probability aggregation regions are mapped to specific conformational regions of the biomacromolecule multiscale complex. Based on the specific conformational region and the heterogeneous nucleation theory, the microscopic contact angle between the primary micro-ice crystals and the specific conformational region is determined. Based on the microscopic contact angle, the heterogeneous nucleation free energy barrier of the specific conformational region is calculated. The thermodynamic evaluation results of the multi-scale complex of the biomacromolecule are obtained based on the heterogeneous nucleation free energy barrier.

[0008] The method provided in this disclosure acquires a process image of the formation of a newly formed ice crystal front based on a preset third observation condition, extracts the advancement contour line of the newly formed ice crystal front from the process image based on a spatial reference, and determines the differentiated time trajectory path of the newly formed ice crystal front in a preset region according to the nucleation spatial probability hotspot map. The newly formed ice crystal front is obtained by a phase transition reaction of a multi-scale complex of biological macromolecules, and the spatial reference is obtained from the core nucleation active centers identified in the thermodynamic evaluation results, including: Based on the preset third observation conditions, pre-cooled water vapor is introduced into the space where the biomacromolecule multiscale complex is located to stimulate the phase transition reaction of the biomacromolecule multiscale complex and obtain the newly formed ice crystal front. The process image of the phase change reaction is obtained, and the process image is analyzed frame by frame to extract the advancing contour line of the newly formed ice crystal front during the phase change reaction process; Using the core nucleation active center identified in the thermodynamic evaluation results as a spatial reference, the differentiated time trajectory path of the newly formed ice crystal front as it slides through a preset region is determined based on the nucleation spatial probability hotspot map. The preset region includes an inactive background bearing area and a specific active region that spans multi-scale complexes.

[0009] The method provided in this disclosure, based on the differentiated time-track path and the propagation contour, obtains a spatiotemporal map; determines the order-of-magnitude transition of the catalytic rate of the newly formed ice crystal front based on the spatiotemporal map; and obtains the quantitative result of the extreme catalytic efficiency induced by the multi-scale complex of biomacromolecules for non-classical crystallization based on the order-of-magnitude transition of the catalytic rate, including: The spatial location information of the newly formed ice crystal front in the differentiated time trajectory path is fused and superimposed with the edge morphology information of the newly formed ice crystal front in the propulsion contour line to obtain a spatiotemporal map. Based on the spatiotemporal map, for each pair of adjacent frame process images, the absolute displacement scale of the newly formed ice crystal front advancing along the normal direction is extracted, along with the time interval corresponding to the absolute displacement scale. The displacement-time series dataset of the phase change reaction process is obtained by continuously extracting the process images frame by frame. The transient dynamic propulsion rate of the newly formed ice crystal front is calculated based on the displacement-time series dataset. Based on the specific active region of the multi-scale complex in the preset region and the transient dynamic propulsion rate, the catalytic rate order-of-magnitude transition of the newly formed ice crystal front is extracted. Using the order-of-magnitude jump in the catalytic rate as a quantitative indicator, the quantitative results of the extreme catalytic efficiency induced by the multi-scale complex of the biomacromolecule to induce non-classical crystallization were obtained.

[0010] This disclosure also provides an apparatus for determining the catalytic efficacy of artificial ice core materials, the apparatus comprising: The acquisition module is used to acquire the sample tilt angle and projection sequence of the biomacromolecule multiscale complex according to the preset first observation conditions, reconstruct and decode the sample tilt angle and projection sequence to obtain the three-dimensional reconstruction map and lattice mismatch of the biomacromolecule multiscale complex, wherein the biomacromolecule multiscale complex is embedded in an ice carrier. The analysis module is used to determine the observation field of view based on the three-dimensional reconstruction map and the lattice mismatch, observe the nascent micro ice crystals according to the observation field of view and the preset second observation conditions and draw the nucleation spatial probability hotspot map, analyze the nucleation spatial probability hotspot map, and obtain the thermodynamic evaluation result of the biomacromolecule multiscale complex, wherein the nascent micro ice crystals are obtained by the biomacromolecule multiscale complex under preset processing. The extraction module is used to acquire process images of the formation of new ice crystal fronts based on preset third observation conditions, extract the advancing contour lines of the new ice crystal fronts from the process images based on spatial references, and determine the differentiated time trajectory paths of the new ice crystal fronts in preset regions according to the nucleation spatial probability hotspot map, wherein the new ice crystal fronts are obtained by phase transition reactions of multi-scale complexes of biological macromolecules, and the spatial references are obtained by the core nucleation active centers identified in the thermodynamic evaluation results; The determination module is used to obtain a spatiotemporal map based on the differentiated time trajectory path and the propulsion contour line, determine the order-of-magnitude transition of the catalytic rate of the newly formed ice crystal front based on the spatiotemporal map, and obtain the quantitative result of the extreme catalytic efficiency induced by the multi-scale complex of the biomacromolecule to induce non-classical crystallization based on the order-of-magnitude transition of the catalytic rate.

[0011] The apparatus provided in this disclosure, wherein the acquisition module is used for: Based on the environmental and equipment parameters in the preset first observation conditions, a series of tilt imaging was performed on the biomacromolecule multiscale complex using cryo-electron tomography to obtain the sample tilt angle and projection sequence of the biomacromolecule multiscale complex. The common structural features in each projection image of the series of tilt imaging are extracted. The sample tilt angle is aligned with the projection sequence based on the common structural features to obtain the alignment result. The alignment result is then reconstructed by back projection to obtain the three-dimensional electron density map of the biomacromolecule multiscale complex in the ice carrier. The three-dimensional electron density map is decoded to obtain the three-dimensional multipolymer conformation information of the biomacromolecule multiscale complex in the ice carrier, and a three-dimensional reconstruction map is drawn based on the three-dimensional multipolymer conformation information. Based on the three-dimensional reconstructed image, a confined region high-resolution imaging of the phase interface between the biomacromolecule multiscale complex and the ice carrier is performed, local fast Fourier transform is extracted, and the lattice mismatch of the biomacromolecule multiscale complex is calculated.

[0012] The apparatus provided in this disclosure, wherein the analysis module is used for: Based on the three-dimensional reconstruction image and lattice mismatch, the observation field of view is determined. Based on the observation field of view and the preset second observation conditions, the ice carrier containing the biomacromolecule multiscale complex is induced to perform in-situ micro-sublimation, exposing the characteristic surface and interface of the biomacromolecule multiscale complex. Based on the characteristic interface and the preset second observation conditions, targeted resublimation is performed on the biomacromolecule multiscale complex to obtain nascent micro ice crystals; Based on the three-dimensional reconstruction map, the nascent micro ice crystals are anchored in three-dimensional space coordinates, and a nucleation space probability hotspot map is drawn. The distribution of the nascent micro ice crystals is determined by analyzing the nucleation space probability hotspot map. Based on the distribution, high-probability aggregation regions of nascent micro ice crystals are extracted from the nucleation space probability hotspot map, and the high-probability aggregation regions are mapped to specific conformational regions of the biomacromolecule multiscale complex. Based on the specific conformational region and the heterogeneous nucleation theory, the microscopic contact angle between the primary micro-ice crystals and the specific conformational region is determined. Based on the microscopic contact angle, the heterogeneous nucleation free energy barrier of the specific conformational region is calculated. The thermodynamic evaluation results of the multi-scale complex of the biomacromolecule are obtained based on the heterogeneous nucleation free energy barrier.

[0013] The apparatus provided in this disclosure, wherein the extraction module is used for: Based on the preset third observation conditions, pre-cooled water vapor is introduced into the space where the biomacromolecule multiscale complex is located to stimulate the phase transition reaction of the biomacromolecule multiscale complex and obtain the newly formed ice crystal front. The process image of the phase change reaction is obtained, and the process image is analyzed frame by frame to extract the advancing contour line of the newly formed ice crystal front during the phase change reaction process; Using the core nucleation active center identified in the thermodynamic evaluation results as a spatial reference, the differentiated time trajectory path of the newly formed ice crystal front as it slides through a preset region is determined based on the nucleation spatial probability hotspot map. The preset region includes an inactive background bearing area and a specific active region that spans multi-scale complexes.

[0014] The apparatus provided in this disclosure, wherein the determining module is used for: The spatial location information of the newly formed ice crystal front in the differentiated time trajectory path is fused and superimposed with the edge morphology information of the newly formed ice crystal front in the propulsion contour line to obtain a spatiotemporal map. Based on the spatiotemporal map, for each pair of adjacent frame process images, the absolute displacement scale of the newly formed ice crystal front advancing along the normal direction is extracted, along with the time interval corresponding to the absolute displacement scale. The displacement-time series dataset of the phase change reaction process is obtained by continuously extracting the process images frame by frame. The transient dynamic propulsion rate of the newly formed ice crystal front is calculated based on the displacement-time series dataset. Based on the specific active region of the multi-scale complex in the preset region and the transient dynamic propulsion rate, the catalytic rate order-of-magnitude transition of the newly formed ice crystal front is extracted. Using the order-of-magnitude jump in the catalytic rate as a quantitative indicator, the quantitative results of the extreme catalytic efficiency induced by the multi-scale complex of the biomacromolecule to induce non-classical crystallization were obtained.

[0015] The technical solution provided in this disclosure has the following advantages compared with the prior art: The method for determining the catalytic efficiency of artificial ice nuclei provided in this disclosure involves obtaining a biomacromolecule multiscale complex based on a multiscale complex. The method acquires the sample tilt angle and projection sequence of the biomacromolecule multiscale complex under preset first observation conditions, and reconstructs and decodes it to obtain a three-dimensional reconstruction image. Based on the three-dimensional reconstruction image, the biomacromolecule multiscale complex is processed under preset second observation conditions to obtain nascent micro-ice crystals and plot a nucleation spatial probability hotspot map. The thermodynamic evaluation results are analyzed. A phase transition reaction of the biomacromolecule multiscale complex is activated under preset third observation conditions. Process images are acquired, and the advancing contour line of the nascent ice crystal front is extracted. The differentiated time-track path is determined by combining the nucleation spatial probability hotspot map. A spatiotemporal map is constructed from the differentiated time-track path and the advancing contour line to determine the order-of-magnitude leap in the catalytic rate of the biomacromolecule multiscale complex, thus obtaining the quantitative result of the extreme catalytic efficiency of the biomacromolecule multiscale complex inducing non-classical crystallization. It achieves faithful analysis of the native hydration folding conformation of multi-scale biomacromolecule complexes, accurately locates and quantifies the core nucleation active center, provides a quantitative reverse-engineering mechanism for interfacial polarity and thermodynamic nucleation barrier, and quantifies the rapid crystallization rate under extreme non-equilibrium conditions in situ. Attached Figure Description

[0016] The above and other features, advantages, and aspects of the embodiments of this disclosure will become more apparent from the accompanying drawings and the following detailed description. Throughout the drawings, the same or similar reference numerals denote the same or similar elements. It should be understood that the drawings are schematic, and the originals and elements are not necessarily drawn to scale.

[0017] Figure 1 A schematic flowchart illustrating the method for determining the catalytic efficacy of artificial ice core materials provided in this embodiment of the disclosure; Figure 2 A schematic diagram of the apparatus for determining the catalytic efficacy of artificial ice core materials provided in this embodiment of the present disclosure. Detailed Implementation

[0018] Embodiments of this disclosure will now be described in more detail with reference to the accompanying drawings. While some embodiments of this disclosure are shown in the drawings, it should be understood that this disclosure can be implemented in various forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided to provide a more thorough and complete understanding of this disclosure. It should be understood that the accompanying drawings and embodiments of this disclosure are for illustrative purposes only and are not intended to limit the scope of protection of this disclosure.

[0019] It should be understood that the steps described in the method embodiments of this disclosure may be performed in different orders and / or in parallel. Furthermore, the method embodiments may include additional steps and / or omit the steps shown. The scope of this disclosure is not limited in this respect.

[0020] The term "comprising" and its variations as used herein are open-ended inclusions, meaning "including but not limited to". The term "based on" means "at least partially based on". The term "one embodiment" means "at least one embodiment"; the term "another embodiment" means "at least one additional embodiment"; the term "some embodiments" means "at least some embodiments". Definitions of other terms will be given in the description below.

[0021] It should be noted that the concepts of "first" and "second" mentioned in this disclosure are used only to distinguish different devices, modules or units, and are not used to limit the order of functions performed by these devices, modules or units or their interdependencies.

[0022] It should be noted that the terms "a" and "a plurality of" used in this disclosure are illustrative rather than restrictive, and those skilled in the art should understand that, unless otherwise expressly indicated in the context, they should be understood as "one or more".

[0023] The names of messages or information exchanged between multiple devices in the embodiments of this disclosure are for illustrative purposes only and are not intended to limit the scope of such messages or information.

[0024] To address the aforementioned issues, this disclosure provides a method for determining the catalytic efficacy of artificial ice core materials. The method will be described below with reference to specific embodiments.

[0025] Figure 1 This is a flowchart illustrating a method for determining the catalytic efficacy of an artificial ice core material according to an embodiment of this disclosure. The method can be executed by a device for determining the catalytic efficacy of an artificial ice core material, wherein the device can be implemented using software and / or hardware, and is generally integrated into an electronic device.

[0026] Example 1: A method for determining the catalytic efficacy of an artificial ice core material provided in this embodiment of the present disclosure includes: S101: Obtain the sample tilt angle and projection sequence of the biomacromolecule multiscale complex according to the preset first observation conditions, reconstruct and decode the sample tilt angle and projection sequence to obtain the three-dimensional reconstruction map and lattice mismatch of the biomacromolecule multiscale complex, wherein the biomacromolecule multiscale complex is embedded in an ice carrier. S102: Determine the observation field of view based on the three-dimensional reconstruction map and lattice mismatch, observe the nascent micro ice crystals according to the observation field of view and the preset second observation conditions, draw the nucleation spatial probability hotspot map, analyze the nucleation spatial probability hotspot map, and obtain the thermodynamic evaluation result of the biomacromolecule multiscale complex, wherein the nascent micro ice crystals are obtained by the biomacromolecule multiscale complex under preset processing. S103: Obtain a process image of the formation of a new ice crystal front based on a preset third observation condition; extract the advancing contour line of the new ice crystal front from the process image based on a spatial reference; determine the differentiated time trajectory path of the new ice crystal front in a preset region according to the nucleation spatial probability hotspot map; wherein the new ice crystal front is obtained by a phase transition reaction of a multi-scale complex of biological macromolecules; and the spatial reference is obtained by the core nucleation active center identified in the thermodynamic evaluation results. S104: A spatiotemporal map is obtained based on the differentiated time trajectory path and the propulsion contour line. The catalytic rate order of magnitude transition of the newly formed ice crystal front is determined based on the spatiotemporal map. Based on the catalytic rate order of magnitude transition, the quantitative result of the extreme catalytic efficiency induced by the multi-scale complex of the biomacromolecule to non-classical crystallization is obtained.

[0027] In this embodiment, a multi-scale complex is obtained and dropped onto a microporous carbon film cryo-electron microscope (cryo-EM) grid, which has undergone hydrophilic treatment. The multi-scale complex is initially prepared in a pretreatment environment to obtain the initial preparation result. The initial preparation result is then cooled using a cryostat to obtain a multi-scale biomolecule complex embedded in an ice carrier. The multi-scale complex is an active substance in a polydisperse suspension or crude extract containing an ice core active component. The pretreatment includes initial preparation of the multi-scale complex and cryostat freezing. After pretreatment, the multi-scale complex... A multi-scale complex of biomacromolecules was obtained, which was embedded under an ice support. A microporous carbon film cryo-electron microscopy grid, a specialized grid for cryo-electron microscopy imaging, was coated with a carbon film with a regular array of micropores. The micropore region was used to form an ultrathin liquid film and embed the sample, while the carbon film framework region was used to support and trap large-scale aggregates. Initial preparation was carried out in a constant temperature and humidity environment. Excess liquid was absorbed from the grid loaded with the multi-scale complex dispersion using filter paper, allowing large-scale aggregates to be trapped in the carbon film framework region, naturally forming a layer no thicker than 100 μm in the micropore region. The process of creating an ultrathin hydrated liquid membrane (nm) involves the initial preparation of a carrier sample, which serves as the initial preparation result. An ultrathin hydrated liquid membrane of suitable thickness is formed within the microporous region. The liquid membrane contains nanoscale biomacromolecules and multi-scale complexes. Large-scale aggregates are retained in the carbon membrane framework region. The freezing process involves dropping the carrier corresponding to the initial preparation result into liquid ethane at a set temperature not exceeding -180℃ at a speed of milliseconds, causing the structural water in the liquid membrane to instantly form amorphous glassy ice, thus obtaining an ice carrier.

[0028] In this embodiment, the preset first observation condition is the sum of environmental and equipment parameters preset to obtain the sample tilt angle and projection sequence. The main process is to transfer the flash-vitrified grid, which has been frozen, into an advanced cryogenic transmission electron microscope equipped with a cold field emission electron gun (Cold-FEG) and an Omega energy filter under the protection of liquid nitrogen temperature throughout the process, and maintain an extremely low temperature observation environment of -196°C.

[0029] In this embodiment, the sample tilt angle is the tilt angle of the carrier relative to the electron beam incident direction during a series of tilt imaging processes. Taking the perpendicular incident electron beam as the zero-degree reference, the carrier is tilted at a certain angle in both the positive and negative directions to obtain projection information of different orientations of the multi-scale complex of biomacromolecules embedded under the ice carrier.

[0030] In this embodiment, the projection sequence is a set of two-dimensional projection images acquired during a series of tilt imaging processes, arranged in order of tilt angle. Each image represents a two-dimensional projection of the electron beam onto the biomacromolecule multiscale complex at the tilt angle, containing superposition information of the internal structure of the biomacromolecule multiscale complex in different directions.

[0031] In this embodiment, reconstruction involves extracting common structural features from each projected image in a series of tilt imaging. Based on these common structural features, the sample tilt angle is aligned with the projection sequence to eliminate mechanical drift and focusing deviation during the tilting process, resulting in an alignment result. The alignment result is then reconstructed using a weighted backprojection algorithm based on the central section theorem. This reconstructs the multi-angle two-dimensional projection of the biomacromolecule multiscale complex into Fourier space, filling it with the original angles. An inverse Fourier transform is then performed to obtain the three-dimensional electron density map of the biomacromolecule multiscale complex. The three-dimensional electron density map is decoded to obtain the three-dimensional multipolymer conformation information of the biomacromolecule multiscale complex in the ice carrier. Based on this conformation information, a three-dimensional reconstruction map is drawn. This reconstruction map is a visual image of the three-dimensional spatial structure of the biomacromolecule multiscale complex in the ice carrier, containing atomic coordinate models, surface topology, and spatial location information of the active phase interface. The three-dimensional multipolymer conformation information includes the periodic spacing of characteristic amino acids on the surface of the biomacromolecule multiscale complex and the lattice constants of the corresponding crystal planes of the ice crystals derived at the interface.

[0032] In this embodiment, ,in, Indicates lattice mismatch; The periodic spacing of characteristic amino acids on the surface of multiscale complexes of biological macromolecules; This represents the lattice constant of the crystal plane corresponding to the ice crystals that are derived at the interface.

[0033] In this embodiment, the preset second observation conditions are the sum of environmental and equipment parameters preset to achieve in-situ micro-sublimation exposure, including: maintaining the cryo-electron microscope field of view; using an in-situ temperature-controlled sample stage to slowly raise the sample ambient temperature to the range of -160℃ to -140℃ at a set rate, while maintaining pressure and holding for a set time; under the preset second observation conditions, in-situ micro-sublimation of the ice carrier embedding the biomacromolecule multiscale complex is induced, exposing the characteristic interface of the biomacromolecule multiscale complex, followed by targeted re-sublimation, resulting in the in-situ formation of new nascent micro-ice crystals on the characteristic interface. The XYZ three-dimensional spatial coordinates of all nascent micro-ice crystals are statistically mapped onto the three-dimensional reconstruction map of the biomacromolecule multiscale complex to obtain a nucleation spatial probability hotspot map, where high-probability areas are hotspots and low-probability areas are cold spots. Based on the analysis of the nucleation spatial probability hotspot map, the measurement of microscopic contact angles, and the calculation of heterogeneous nucleation free energy barrier, the positions of the core nucleation active centers on the surface of the biomacromolecule multiscale complex and the degree to which the core nucleation active centers reduce the nucleation barrier are obtained, yielding thermodynamic evaluation results. Thermodynamic assessment results include the location of the core nucleation active center and the degree to which the core nucleation active center reduces the nucleation barrier.

[0034] In this embodiment, the preset third observation condition is the sum of environmental and equipment parameters preset to induce a direct gas-solid phase transition reaction. The main operational steps are to maintain the in-situ observation state after determining the three-dimensional spatial coordinates of the core nucleation active center in the multi-scale thermodynamic evaluation results of the complex, and to maintain continuous focusing and alignment of the cryo-electron microscope field of view on the specific conformational region. At the same time, relying on the advanced cryogenic environment sample rod system equipped with a high-precision microfluidic control gas phase flow loop and dual-tilt temperature control function, the in-situ observation platform coupled with gaseous and extreme cold is officially activated, and the vacuum and auxiliary cooling synergistic mechanism is initiated to evacuate the micro-nano space in the sample chamber to the extremely low pressure limit (at 10). 5 The target environment is rapidly reduced to an ultracold limit temperature (approximately 100K) to establish extreme temperature and pressure conditions, providing an extremely non-equilibrium physical driving field for subsequent triggering and measurement of transient dynamic rates. After the environmental parameters stabilize, the high-precision micro-controlled valve of the in-situ sample rod is opened to pulse-directedly introduce a trace amount of high-purity pre-cooled water vapor into the nanoscale confined space of the ultracold vacuum. This instantly constructs a molecular-level concentration and temperature cliff gradient on the surface of the multi-scale complex of biomacromolecules, strongly driving the ultracold water vapor molecules to completely bypass the classical nucleation "gas-liquid (water droplet)-solid" delayed path and directly forcibly excite a non-classical "gas-solid" heterogeneous crystallization phase transition reaction on the calibrated characteristic interface. The moving boundary between the solid ice crystals generated in the gas-solid heterogeneous crystallization phase transition reaction and the surrounding gas phase is the nascent ice crystal front.

[0035] In this embodiment, the process images are obtained using a high-sensitivity direct electron detector at the cryo-electron microscope end with a sub-millisecond time resolution (500–1500). The high-frequency continuous frame sequence images obtained by continuous high-fidelity in-situ dynamic recording of the entire gas-solid crystallization phase transition reaction (fps high-frequency frame rate sampling) are analyzed frame by frame. The real-time edge morphology line of the newly formed ice crystal front is extracted and labeled in each process image to obtain the advancement contour line. With the core nucleation active center as the spatial reference, the differentiated time trajectory path of the newly formed ice crystal front as it slides over surface regions with different properties is extracted, which has significant differences in the rate of change of spatial position. The differentiated time trajectory path is further divided into the uninterrupted advancement path in the non-active background bearing area (preset area) and the targeted catalytic acceleration advancement path in the core nucleation active center area. When the newly formed ice crystal front crosses the core nucleation active center, the transient dynamic advancement rate shows a step-like, order-of-magnitude jump phenomenon, which is the catalytic rate order-of-magnitude jump. The final quantitative conclusion on the extreme catalytic efficiency of non-classical crystallization induced by biomacromolecule multi-scale complex is obtained by calculating and judging the multiple of the catalytic rate order-of-magnitude jump as a quantitative index.

[0036] In this embodiment, the core nucleation active center is a specific groove region on the surface of a multi-scale complex of biomacromolecules with an extremely low heterogeneous nucleation free energy barrier, as determined by thermodynamic evaluation results. It is a functional site that induces premature freezing of supercooled water.

[0037] In this embodiment, the preset region is a pre-divided comparison region based on the nucleation spatial probability hotspot map, including an inactive background bearing region and a specific active region spanning multiple scale complexes.

[0038] In this embodiment, the spatiotemporal map is a three-dimensional visualization data volume that is formed by fusing and superimposing spatial location information and edge morphology information, and simultaneously includes the XY spatial dimension and the T time dimension.

[0039] In this embodiment, a spatiotemporal map is obtained by fusing and superimposing the spatial location information of the differentiated time-track path and the edge morphology information of the propulsion contour. The absolute displacement scale and time interval of the normal direction of the newly formed ice crystal front are extracted frame by frame to obtain a displacement-time series dataset. The transient dynamic propulsion rate is calculated, and the catalytic rate order-of-magnitude transition is extracted based on the specific active region of the multi-scale complex. The transition multiple is used as a quantitative index to obtain the quantification result of the extreme catalytic efficiency.

[0040] The working principle and beneficial effects of the embodiments disclosed herein are as follows: Based on multi-scale complexes, biomacromolecule multi-scale complexes are obtained. The sample tilt angle and projection sequence of the biomacromolecule multi-scale complex are acquired under preset first observation conditions, and a three-dimensional reconstruction map is obtained through reconstruction and decoding. Based on the three-dimensional reconstruction map, the biomacromolecule multi-scale complex is processed under preset second observation conditions to obtain nascent micro-ice crystals and plot a nucleation spatial probability hotspot map. Thermodynamic evaluation results are analyzed. Based on preset third observation conditions, the phase transition reaction of the biomacromolecule multi-scale complex is activated, process images are acquired, and the advancing contour line of the nascent ice crystal front is extracted. Differential time-track paths are determined by combining the nucleation spatial probability hotspot map. A spatiotemporal map is constructed from the differential time-track paths and advancing contour lines to determine the order-of-magnitude leap in the catalytic rate of the biomacromolecule multi-scale complex, obtaining the quantitative results of the extreme catalytic efficiency of non-classical crystallization induced by the biomacromolecule multi-scale complex. This achieves faithful analysis of the original hydration folding conformation of the biomacromolecule multi-scale complex, accurately locates and quantifies the core nucleation active center, provides a quantitative backward deduction mechanism for interface polarity and thermodynamic nucleation barrier, and quantifies the rapid crystallization advancement rate under extreme non-equilibrium conditions in situ.

[0041] Example 2: The method provided in this embodiment of the present disclosure obtains the sample tilt angle and projection sequence of a multi-scale biomolecular complex according to a preset first observation condition, reconstructs and decodes the sample tilt angle and projection sequence to obtain a three-dimensional reconstruction image and lattice mismatch of the multi-scale biomolecular complex, wherein the multi-scale biomolecular complex is embedded in an ice carrier, comprising: Based on the environmental and equipment parameters in the preset first observation conditions, a series of tilt imaging was performed on the biomacromolecule multiscale complex using cryo-electron tomography to obtain the sample tilt angle and projection sequence of the biomacromolecule multiscale complex. The common structural features in each projection image of the series of tilt imaging are extracted. The sample tilt angle is aligned with the projection sequence based on the common structural features to obtain the alignment result. The alignment result is then reconstructed by back projection to obtain the three-dimensional electron density map of the biomacromolecule multiscale complex in the ice carrier. The three-dimensional electron density map is decoded to obtain the three-dimensional multipolymer conformation information of the biomacromolecule multiscale complex in the ice carrier, and a three-dimensional reconstruction map is drawn based on the three-dimensional multipolymer conformation information. Based on the three-dimensional reconstructed image, a confined region high-resolution imaging of the phase interface between the biomacromolecule multiscale complex and the ice carrier is performed, local fast Fourier transform is extracted, and the lattice mismatch of the biomacromolecule multiscale complex is calculated.

[0042] In this embodiment, cryo-electron tomography (cryo-electron tomography) is a technique for performing a series of tilt images on the multi-scale complex of the biomacromolecule under extremely low temperature conditions. During cryo-electron tomography, the sample stage is controlled to tilt stepwise according to a preset angle range and angle step size. One projection image is acquired for each tilt step, resulting in a series of tilt images covering multiple angles. The fixed reference objects or image features that can be identified in each projection image of the series of tilt images are the common structural features. The common structural features may include gold particle markers, specific contrast regions in the ice carrier background, or identifiable structural domains of biomacromolecules. Based on the common structural features, the image set obtained after drift correction and spatial registration of the sample tilt angle and the projection images of each tilt angle in the projection sequence is the alignment result.

[0043] In this embodiment, the three-dimensional electron density map is a spatial electron density distribution map represented by a three-dimensional voxel array, obtained by weighted back projection reconstruction of the alignment results. The gray value of each voxel reflects the scattering ability of the electron beam by the spatial position, and the density of the biomacromolecule region is higher than that of the ice carrier background.

[0044] In this embodiment, the three-dimensional multimer conformation information is the three-dimensional spatial coordinates and assembly relationship of each atom or amino acid residue in the multi-scale complex of the biomacromolecule obtained by decoding.

[0045] The working principle and beneficial effects of the embodiments disclosed herein are as follows: Multi-scale complexes are processed in a pretreatment environment to obtain multi-scale biomolecular complexes embedded in an ice carrier. Based on preset first observation conditions, a series of tilt imaging techniques are performed using cryo-electron tomography to obtain the sample tilt angle and projection sequence of the multi-scale biomolecular complex embedded in the ice carrier. Common structural feature alignment sequences are extracted from these sequences, and the three-dimensional electron density map is reconstructed and decoded via back-projection to obtain three-dimensional polymer conformation information and to draw a three-dimensional reconstructed map and lattice mismatch. This achieves in-situ structural analysis of multi-scale biomolecular complexes under near-physiological conditions and provides direct structural evidence for revealing the multi-scale dynamic assembly mechanism of the complex.

[0046] Example 3: The method provided in this embodiment determines the observation field of view based on the three-dimensional reconstruction image and lattice mismatch, observes primary micro-ice crystals according to the observation field of view and preset second observation conditions, and draws a nucleation spatial probability hotspot map. The nucleation spatial probability hotspot map is analyzed to obtain the thermodynamic evaluation result of the biomacromolecule multi-scale complex, wherein the primary micro-ice crystals are obtained by preset processing of the biomacromolecule multi-scale complex. The preset processing includes in-situ micro-sublimation and targeted re-sublimation, including: Based on the three-dimensional reconstruction image and lattice mismatch, the observation field of view is determined. Based on the observation field of view and the preset second observation conditions, the ice carrier containing the biomacromolecule multiscale complex is induced to perform in-situ micro-sublimation, exposing the characteristic surface and interface of the biomacromolecule multiscale complex. Based on the characteristic interface and the preset second observation conditions, targeted resublimation is performed on the biomacromolecule multiscale complex to obtain nascent micro ice crystals; Based on the three-dimensional reconstruction map, the nascent micro ice crystals are anchored in three-dimensional space coordinates, and a nucleation space probability hotspot map is drawn. The distribution of the nascent micro ice crystals is determined by analyzing the nucleation space probability hotspot map. Based on the distribution, high-probability aggregation regions of nascent micro ice crystals are extracted from the nucleation space probability hotspot map, and the high-probability aggregation regions are mapped to specific conformational regions of the biomacromolecule multiscale complex. Based on the specific conformational region and the heterogeneous nucleation theory, the microscopic contact angle between the primary micro-ice crystals and the specific conformational region is determined. Based on the microscopic contact angle, the heterogeneous nucleation free energy barrier of the specific conformational region is calculated. The thermodynamic evaluation results of the multi-scale complex of the biomacromolecule are obtained based on the heterogeneous nucleation free energy barrier.

[0047] In this embodiment, the observation field of view is the sample area imaged by electron beam irradiation under a specific magnification of the cryo-electron microscope. The low lattice mismatch region, determined by the three-dimensional reconstruction image and the lattice mismatch, is set as the observation field of view. Under in-situ temperature control conditions within the cryo-electron microscope, amorphous glassy ice containing multi-scale biomolecular complexes is heated to -160°C to -140°C and held at pressure for a set time, inducing nanoscale local micro-sublimation of the amorphous glassy ice on the surface, which is called in-situ micro-sublimation. Based on the exposure of the characteristic interfaces of the multi-scale complexes originally buried deep within the ice by in-situ micro-sublimation, the characteristic interfaces are regions with specific physicochemical properties on the surface of the multi-scale biomolecular complexes exposed by in-situ micro-sublimation, which become the physical substrate for subsequent targeted re-sublimation.

[0048] In this embodiment, the local gas phase supersaturation conditions caused by in-situ micro-sublimation are maintained. The trace amount of ultracold water vapor remaining in the electron microscope cavity is used to preferentially sublimate and nucleate water vapor on the exposed characteristic interface, which is a targeted resublimation. Primary micro ice crystals are formed based on the targeted resublimation.

[0049] In this embodiment, the three-dimensional spatial coordinate anchoring is an operation process that uses the three-dimensional reconstruction image as a spatial reference system to measure the spatial position coordinates of each nascent micro-ice crystal captured in the field of view in the three dimensions of the X-axis, Y-axis, and Z-axis, and then maps the positioning of the nascent micro-ice crystals to the three-dimensional structure of the multi-scale complex of biomacromolecules.

[0050] In this embodiment, the distribution refers to the thermodynamic barrier distribution of nascent micro-ice crystals. The frequency of occurrence and spatial aggregation state of nascent micro-ice crystals on the surface of the multi-scale complex of biomacromolecules are assessed based on the nucleation spatial probability hotspot map and classical heterogeneous nucleation theory to obtain the distribution. High-probability aggregation regions are identified by analyzing the distribution; these regions are locations on the nucleation spatial probability hotspot map where the frequency of nascent micro-ice crystal occurrence is significantly higher than in other regions. After mapping these high-probability aggregation regions, specific conformational regions are defined, including characteristic folded grooves, periodic grooves formed by tandem repeating sequences, etc.

[0051] In this embodiment, the heterogeneous nucleation theory is a classical thermodynamic theory describing the nucleation and crystallization of water molecules on a heterogeneous substrate surface. This is mainly reflected in the fact that the heterogeneous substrate can reduce the free energy barrier that pure water needs to overcome for homogeneous nucleation. The degree to which this free energy barrier is reduced depends on the microscopic contact angle formed between the ice nucleus and the substrate.

[0052] In this embodiment, the micro-contact angle is the three-phase contact angle formed between the primary micro-ice crystals and the surface of a specific conformation region during heterogeneous nucleation. The smaller the micro-contact angle value, the better the interfacial wettability and the lower the nucleation barrier.

[0053] In this embodiment, the heterogeneous nucleation free energy barrier is the thermodynamic barrier that needs to be overcome to catalyze the freezing of supercooled water in a specific conformational region, calculated according to classical heterogeneous nucleation theory. The corresponding formula is: ,in, This represents the free energy barrier that must be overcome for heterogeneous nucleation to occur; The free energy barrier represents the homogeneous nucleation of pure water; This represents the microscopic contact angle at the interface between the ice nucleus and a specific substrate.

[0054] The working principle and beneficial effects of the embodiments disclosed herein are as follows: The observation field is determined based on a three-dimensional reconstruction image and lattice mismatch. In-situ micro-sublimation of the ice carrier is induced according to a preset second observation condition to expose characteristic interfaces. Targeted re-sublimation is then performed to obtain nascent micro-ice crystals. The nascent micro-ice crystals are anchored in three-dimensional space based on the three-dimensional reconstruction image, and a nucleation spatial probability hotspot map is drawn. The distribution is analyzed, high-probability aggregation regions are extracted and mapped to specific conformational regions. The microscopic contact angle is determined based on the specific conformational region and heterogeneous nucleation theory, and the heterogeneous nucleation free energy barrier is calculated to obtain thermodynamic evaluation results. This achieves quantitative evaluation of the heterogeneous nucleation free energy barrier, revealing the thermodynamic mechanism of ice nucleation induced by the composite. It realizes the quantitative reverse deduction of the microscopic contact angle response and thermodynamic barrier reduction at the nucleation interface, providing an objective spatial and energy distribution standard for accurately calibrating the core nucleation active centers that induce rapid freezing in materials.

[0055] Example 4: The method provided in this embodiment acquires a process image of the formation of a newly formed ice crystal front based on a preset third observation condition, extracts the advancement contour line of the newly formed ice crystal front from the process image based on a spatial reference, and determines the differentiated time trajectory path of the newly formed ice crystal front in a preset region according to the nucleation spatial probability hotspot map. The newly formed ice crystal front is obtained by a phase transition reaction of a multi-scale complex of biological macromolecules, and the spatial reference is obtained from the core nucleation active centers identified in the thermodynamic evaluation results, including: Based on the preset third observation conditions, pre-cooled water vapor is introduced into the space where the biomacromolecule multiscale complex is located to stimulate the phase transition reaction of the biomacromolecule multiscale complex and obtain the newly formed ice crystal front. The process image of the phase change reaction is obtained, and the process image is analyzed frame by frame to extract the advancing contour line of the newly formed ice crystal front during the phase change reaction process; Using the core nucleation active center identified in the thermodynamic evaluation results as a spatial reference, the differentiated time trajectory path of the newly formed ice crystal front as it slides through a preset region is determined based on the nucleation spatial probability hotspot map. The preset region includes an inactive background bearing area and a specific active region that spans multi-scale complexes.

[0056] In this embodiment, the pre-cooled water vapor is a trace amount of high-purity water vapor that has undergone pre-cooling treatment. The pre-cooled water vapor is introduced into the nano-confined space of extreme cold vacuum through a high-precision micro-controlled valve in a pulse manner. As a reactant of gas-solid phase transition, it bypasses the gas-liquid-solid path under extreme non-equilibrium conditions and directly undergoes direct phase transition crystallization on the target surface to obtain a new ice crystal front.

[0057] In this embodiment, frame-by-frame analysis involves using image processing algorithms to analyze each process image of a high-frequency continuous frame sequence, identifying and extracting the position, shape, and boundary information of the newly formed ice crystal front in each process image.

[0058] In this embodiment, the inactive background support region is the background region around the multi-scale complex of biomacromolecules that does not have nucleation catalytic activity, such as the region of a blank pure carbon film; crossing the specific active region of the multi-scale complex is the process by which the newly formed ice crystal front advances from the inactive background support region into and passes through the region where the core nucleation active center is located.

[0059] The working principle and beneficial effects of this embodiment are as follows: Pre-cooled water vapor is introduced based on a preset third observation condition to induce a phase change reaction and obtain a newly formed ice crystal front. Process images are acquired, and the advancement contour of the newly formed ice crystal front is extracted frame by frame. Using the core nucleation active center identified by the thermodynamic evaluation results as a spatial reference, the differentiated time-track paths of the newly formed ice crystal front across the inactive background bearing area and across specific active regions of the multi-scale complex are determined based on the nucleation spatial probability hotspot map. This enables differentiated tracking of the time-track paths of the newly formed ice crystal front in the active and inactive regions, calibrates catalytic sites, and provides a spatial reference for the kinetic efficiency quantification.

[0060] Example 5: The method provided in this embodiment of the present disclosure obtains a spatiotemporal map based on the differentiated time-track path and the propulsion profile, determines the order-of-magnitude transition of the catalytic rate of the newly formed ice crystal front based on the spatiotemporal map, and obtains the quantitative result of the extreme catalytic efficiency induced by the multi-scale complex of the biomacromolecule to induce non-classical crystallization based on the order-of-magnitude transition of the catalytic rate, including: The spatial location information of the newly formed ice crystal front in the differentiated time trajectory path is fused and superimposed with the edge morphology information of the newly formed ice crystal front in the propulsion contour line to obtain a spatiotemporal map. Based on the spatiotemporal map, for each pair of adjacent frame process images, the absolute displacement scale of the newly formed ice crystal front advancing along the normal direction is extracted, along with the time interval corresponding to the absolute displacement scale. The displacement-time series dataset of the phase change reaction process is obtained by continuously extracting the process images frame by frame. The transient dynamic propulsion rate of the newly formed ice crystal front is calculated based on the displacement-time series dataset. Based on the specific active region of the multi-scale complex in the preset region and the transient dynamic propulsion rate, the catalytic rate order-of-magnitude transition of the newly formed ice crystal front is extracted. Using the order-of-magnitude jump in the catalytic rate as a quantitative indicator, the quantitative results of the extreme catalytic efficiency induced by the multi-scale complex of the biomacromolecule to induce non-classical crystallization were obtained.

[0061] In this embodiment, the spatial location information is the three-dimensional spatial coordinate data of the newly formed ice crystal front in each frame of the process image in the differentiated time track path, which describes the position of the newly formed ice crystal front in the X-axis, Y-axis and Z-axis directions as time changes.

[0062] In this embodiment, edge morphology information is obtained based on the contour shape and boundary geometric features of the newly formed ice crystal front in each frame of the process image in the advance contour line. This information is used to describe the spatial occupancy range and front curvature change of the newly formed ice crystal front at each time step.

[0063] In this embodiment, in the spatiotemporal map, for each pair of adjacent process images, the actual spatial distance of the newly formed ice crystal front advancing along the corresponding normal direction from the position of the previous frame to the position of the next frame is measured. This actual spatial distance is the absolute displacement scale, and the sampling time difference between each pair of adjacent process images is the time interval. A displacement-time series dataset is obtained based on all absolute displacement scales and time intervals. The instantaneous advancement velocity of the newly formed ice crystal front along the corresponding normal direction between adjacent frames is calculated based on the displacement-time series dataset, using the corresponding formula: ,in, dL represents the transient kinetic propulsion rate that facilitates crystallization; dL represents the absolute displacement scale of the normal propulsion of the ice crystal front between adjacent microsecond image frames; dt represents the sampling interval timestamp. The transient kinetic propulsion rate is the baseline value in the inactive background bearing region and increases abruptly when crossing specific active regions of the multi-scale complex.

[0064] In this embodiment, the quantitative index is the order-of-magnitude jump in catalytic rate, and the catalytic efficiency is characterized by the order-of-magnitude increase in transient kinetic propulsion rate in the active region relative to the inactive background region.

[0065] The working principle and beneficial effects of this embodiment are as follows: A spatiotemporal map is obtained by fusing and superimposing the spatial location information of differentiated time-track paths and the edge morphology information of the propulsion contour line. The absolute displacement scale and time interval of the normal direction of the newly formed ice crystal front are extracted frame by frame to obtain a displacement-time series dataset. The transient dynamic propulsion rate is calculated, and catalytic rate order-of-magnitude transitions are extracted based on specific active regions of the multi-scale complex. The transition factor is used as a quantification index to obtain the quantification result of extreme catalytic efficiency. This achieves a comprehensive understanding of the mechanism from the molecular-level catalytic source to macroscopic explosive interphase transitions, providing a directly applicable microscopic diagnostic framework for the design and screening of highly efficient biological / biomimetic ice core materials.

[0066] In this embodiment, the corresponding example is the preparation of a polydisperse suspension by dispersing Snomax active biological ice nucleoprotein powder in ultrapure water. An appropriate amount of the suspension is dropped onto a microporous carbon membrane support with a pore size of 1.2 μm, which has undergone plasma glow discharge hydrophilization treatment. In a constant temperature and humidity environment of 22°C and relative humidity greater than 90%, excess liquid is precisely absorbed using filter paper, resulting in an ultrathin hydrated liquid film with a thickness not exceeding 100 nm in the microporous region. The support is then rapidly vitrified by ejecting it into liquid ethane at -185°C using a rapid immersion cryostat. The frozen sample is transferred under liquid nitrogen protection throughout the process and loaded into a cryo-transmission electron microscope equipped with a cold field emission electron gun and an Omega energy filter, maintaining an extremely low temperature observation environment of -196°C and activating the low-radiation-dose imaging protection mode. A 3D reconstruction of the Snomax protein molecule was obtained through cryo-electron tomography combined with single-particle analysis and decoding, revealing a highly ordered β-helical aggregated bundled conformation. High-resolution imaging of a confined region was performed on the ice carrier closely adhering to the protein surface. Local fast Fourier transforms were extracted to obtain the lattice constants of the characteristic amino acid periodic spacing on the Snomax protein surface and the corresponding crystal planes of the ice carrier. The lattice mismatch was calculated using the lattice matching formula. Based on the 3D reconstruction and lattice mismatch, the observation field was determined. The sample environment temperature was uniformly and slowly raised to -142℃ using an in-situ temperature-controlled sample stage, and in-situ micro-sublimation was performed for approximately 3 minutes under pressure to expose the characteristic interface. The sample was then recooled to -196℃ to stabilize the solid-phase interface. Targeted resublimation was used to induce the formation of primary micro-ice crystals, and over 800 primary micro-ice crystals were captured through continuous scanning. 3D spatial coordinate anchoring was performed, and a nucleation spatial probability hotspot map was plotted. Analysis revealed that nearly 92% of the nascent microcrystals were not randomly distributed, but precisely aggregated in the core groove of the high-density β-helical tandem repeat sequence of the Snomax protein. By mapping the high-probability aggregation region to a specific conformational region, measuring the microscopic contact angle, and substituting it into the classical heterogeneous nucleation theory formula, the heterogeneous nucleation free energy barrier was calculated. Thermodynamic assessment results showed that the microscopic contact angle of the core groove region (specific conformational region) approached zero, and the nucleation free energy barrier was significantly reduced, thermodynamically confirming that the core groove region is the core nucleation active center. To maintain continuous focusing and alignment of the cryo-electron microscopy field on the core nucleation active center, a dual-tilt cryogenic sample holder system with a microfluidic gas phase loop was activated. A vacuum and auxiliary cooling synergistic mechanism was initiated, evacuating the target area's micro-nano space to the order of 1.5 × 10⁻⁵ Pa and drastically reducing the temperature to the ultracold limit of 100 K, establishing an extremely non-equilibrium physical driving field.After the environmental parameters stabilize, a high-precision micro-controlled valve is opened to introduce a trace amount of high-purity pre-cooled water vapor in a pulsed, directional manner. This instantly constructs a molecular-level concentration and temperature gradient at the characteristic interface of the Snomax protein, strongly driving the ultracooled water vapor molecules to bypass the gas-liquid-solid path and directly induce a non-classical gas-solid heterogeneous crystallization phase transition reaction in the β-helical active target region. Simultaneously, a high-sensitivity direct electron detector is activated to continuously and dynamically record the entire phase transition reaction process at a high frame rate of 1200 fps, acquiring process images. Image processing algorithms are used to analyze the process images frame by frame, extracting the advancement contour of the newly formed ice crystal front. Using the core nucleation active center identified by thermodynamic evaluation results as a spatial reference, the differentiated temporal trajectory paths of the newly formed ice crystal front in the inactive background carrying region and across specific active regions of the Snomax multi-scale complex are determined based on the nucleation spatial probability hotspot map. The spatial location information of the differentiated temporal trajectory paths is fused and superimposed with the edge morphology information of the advancement contour to obtain a spatiotemporal map. The absolute displacement scale and corresponding time interval of the normal advance of the newly formed ice crystal front were extracted frame by frame to obtain a displacement-time series dataset. Substituting this data into the interface propagation kinetic rate equation, it was found that when the newly formed ice crystal front advances within the background region of the inactive carbon film, a significant pause of nearly 0.5 seconds occurs every tens of nanometers. When the same newly formed ice crystal front crosses the core nucleation active center, the transient kinetic propagation rate exhibits an order-of-magnitude jump in catalytic rate, instantly increasing to approximately 500 times that of the background region. Using the order-of-magnitude jump as a quantitative indicator, the quantitative results of the extreme catalytic efficiency of non-classical crystallization induced by the Snomax multi-scale complex were obtained.

[0067] To achieve the above embodiments, this disclosure also proposes an apparatus for determining the catalytic efficiency of artificial ice core materials.

[0068] Figure 2 This is a schematic diagram of the device for determining the catalytic efficacy of artificial ice core materials provided in an embodiment of this disclosure. The device 200 can be implemented by software and / or hardware and is generally integrated into an electronic device. Figure 2 As shown, the device 200 includes: an acquisition module 201, an analysis module 202, an extraction module 203, and a determination module 204, wherein, The acquisition module 201 is used to acquire the sample tilt angle and projection sequence of the biomacromolecule multiscale complex according to the preset first observation conditions, reconstruct and decode the sample tilt angle and projection sequence to obtain the three-dimensional reconstruction map and lattice mismatch of the biomacromolecule multiscale complex, wherein the biomacromolecule multiscale complex is embedded in an ice carrier. Analysis module 202 is used to determine the observation field of view based on the three-dimensional reconstruction map and lattice mismatch, observe the nascent micro ice crystals according to the observation field of view and the preset second observation conditions and draw the nucleation spatial probability hotspot map, analyze the nucleation spatial probability hotspot map, and obtain the thermodynamic evaluation result of the biomacromolecule multiscale complex, wherein the nascent micro ice crystals are obtained by the biomacromolecule multiscale complex under preset processing. Extraction module 203 is used to acquire process images of the formation of new ice crystal fronts based on preset third observation conditions, extract the advancing contour lines of the new ice crystal fronts from the process images based on spatial references, and determine the differentiated time trajectory path of the new ice crystal fronts in preset regions according to the nucleation spatial probability hotspot map, wherein the new ice crystal fronts are obtained by phase transition reactions of multi-scale complexes of biological macromolecules, and the spatial references are obtained by the core nucleation active centers identified in the thermodynamic evaluation results; The determination module 204 is used to obtain a spatiotemporal map based on the differentiated time trajectory path and the propulsion contour line, determine the order-of-magnitude transition of the catalytic rate of the newly formed ice crystal front based on the spatiotemporal map, and obtain the quantitative result of the extreme catalytic efficiency induced by the multi-scale complex of the biomacromolecule to induce non-classical crystallization based on the order-of-magnitude transition of the catalytic rate.

[0069] The apparatus provided in this disclosure, wherein the acquisition module 201 is used for: Based on the environmental and equipment parameters in the preset first observation conditions, a series of tilt imaging was performed on the biomacromolecule multiscale complex using cryo-electron tomography to obtain the sample tilt angle and projection sequence of the biomacromolecule multiscale complex. The common structural features in each projection image of the series of tilt imaging are extracted. The sample tilt angle is aligned with the projection sequence based on the common structural features to obtain the alignment result. The alignment result is then reconstructed by back projection to obtain the three-dimensional electron density map of the biomacromolecule multiscale complex in the ice carrier. The three-dimensional electron density map is decoded to obtain the three-dimensional multipolymer conformation information of the biomacromolecule multiscale complex in the ice carrier, and a three-dimensional reconstruction map is drawn based on the three-dimensional multipolymer conformation information. Based on the three-dimensional reconstructed image, a confined region high-resolution imaging of the phase interface between the biomacromolecule multiscale complex and the ice carrier is performed, local fast Fourier transform is extracted, and the lattice mismatch of the biomacromolecule multiscale complex is calculated.

[0070] The apparatus provided in this disclosure, wherein the analysis module 202 is used for: Based on the three-dimensional reconstruction image and lattice mismatch, the observation field of view is determined. Based on the observation field of view and the preset second observation conditions, the ice carrier containing the biomacromolecule multiscale complex is induced to perform in-situ micro-sublimation, exposing the characteristic surface and interface of the biomacromolecule multiscale complex. Based on the characteristic interface and the preset second observation conditions, targeted resublimation is performed on the biomacromolecule multiscale complex to obtain nascent micro ice crystals; Based on the three-dimensional reconstruction map, the nascent micro ice crystals are anchored in three-dimensional space coordinates, and a nucleation space probability hotspot map is drawn. The distribution of the nascent micro ice crystals is determined by analyzing the nucleation space probability hotspot map. Based on the distribution, high-probability aggregation regions of nascent micro ice crystals are extracted from the nucleation space probability hotspot map, and the high-probability aggregation regions are mapped to specific conformational regions of the biomacromolecule multiscale complex. Based on the specific conformational region and the heterogeneous nucleation theory, the microscopic contact angle between the primary micro-ice crystals and the specific conformational region is determined. Based on the microscopic contact angle, the heterogeneous nucleation free energy barrier of the specific conformational region is calculated. The thermodynamic evaluation results of the multi-scale complex of the biomacromolecule are obtained based on the heterogeneous nucleation free energy barrier.

[0071] The apparatus provided in this disclosure, wherein the extraction module 203 is used for: Based on the preset third observation conditions, pre-cooled water vapor is introduced into the space where the biomacromolecule multiscale complex is located to stimulate the phase transition reaction of the biomacromolecule multiscale complex and obtain the newly formed ice crystal front. The process image of the phase change reaction is obtained, and the process image is analyzed frame by frame to extract the advancing contour line of the newly formed ice crystal front during the phase change reaction process; Using the core nucleation active center identified in the thermodynamic evaluation results as a spatial reference, the differentiated time trajectory path of the newly formed ice crystal front as it slides through a preset region is determined based on the nucleation spatial probability hotspot map. The preset region includes an inactive background bearing area and a specific active region that spans multi-scale complexes.

[0072] The apparatus provided in this disclosure, wherein the determining module 204 is configured to: The spatial location information of the newly formed ice crystal front in the differentiated time trajectory path is fused and superimposed with the edge morphology information of the newly formed ice crystal front in the propulsion contour line to obtain a spatiotemporal map. Based on the spatiotemporal map, for each pair of adjacent frame process images, the absolute displacement scale of the newly formed ice crystal front advancing along the normal direction is extracted, along with the time interval corresponding to the absolute displacement scale. The displacement-time series dataset of the phase change reaction process is obtained by continuously extracting the process images frame by frame. The transient dynamic propulsion rate of the newly formed ice crystal front is calculated based on the displacement-time series dataset. Based on the specific active region of the multi-scale complex in the preset region and the transient dynamic propulsion rate, the catalytic rate order-of-magnitude transition of the newly formed ice crystal front is extracted. Using the order-of-magnitude jump in the catalytic rate as a quantitative indicator, the quantitative results of the extreme catalytic efficiency induced by the multi-scale complex of the biomacromolecule to induce non-classical crystallization were obtained.

[0073] The apparatus for determining the catalytic efficacy of artificial ice nuclei provided in this disclosure can execute the method for determining the catalytic efficacy of artificial ice nuclei provided in any embodiment of this disclosure, and has the corresponding functional modules and beneficial effects for executing the method.

[0074] To implement the above embodiments, this disclosure also proposes a computer program product, including a computer program / instructions, which, when executed by a processor, implements the method for determining the catalytic efficiency of the artificial ice core material in the above embodiments.

[0075] To implement the above embodiments, this disclosure also proposes an electronic device comprising: a processor; a memory for storing executable instructions of the processor; and a processor for reading the executable instructions from the memory and executing the instructions to implement the method for determining the catalytic efficacy of artificial ice core materials as provided in the embodiments of this disclosure.

[0076] This disclosure also proposes a computer-readable storage medium storing a computer program for performing a method for determining the catalytic efficacy of an artificial ice core material as provided in this disclosure.

[0077] It should be noted that the computer-readable medium described in this disclosure can be a computer-readable signal medium or a computer-readable storage medium, or any combination thereof. A computer-readable storage medium can be, for example,—but not limited to—an electrical, magnetic, optical, electromagnetic, infrared, or semiconductor system, apparatus, or device, or any combination thereof. More specific examples of a computer-readable storage medium may include, but are not limited to: an electrical connection having one or more wires, a portable computer disk, a hard disk, random access memory (RAM), read-only memory (ROM), erasable programmable read-only memory (EPROM or flash memory), optical fiber, portable compact disk read-only memory (CD-ROM), optical storage device, magnetic storage device, or any suitable combination thereof. In this disclosure, a computer-readable storage medium can be any tangible medium containing or storing a program that can be used by or in conjunction with an instruction execution system, apparatus, or device. In this disclosure, a computer-readable signal medium can include a data signal propagated in baseband or as part of a carrier wave, carrying computer-readable program code. Such propagated data signals can take various forms, including but not limited to electromagnetic signals, optical signals, or any suitable combination thereof. A computer-readable signal medium can be any computer-readable medium other than a computer-readable storage medium, which can send, propagate, or transmit a program for use by or in connection with an instruction execution system, apparatus, or device. The program code contained on the computer-readable medium can be transmitted using any suitable medium, including but not limited to: wires, optical fibers, RF (radio frequency), etc., or any suitable combination thereof.

[0078] In some implementations, clients and servers can communicate using any currently known or future-developed network protocol such as HTTP (Hypertext Transfer Protocol) and can interconnect with digital data communication (e.g., communication networks) of any form or medium. Examples of communication networks include local area networks (“LANs”), wide area networks (“WANs”), the Internet (e.g., the Internet of Things), and peer-to-peer networks (e.g., ad hoc peer-to-peer networks), as well as any currently known or future-developed networks.

[0079] The aforementioned computer-readable medium may be included in the aforementioned electronic device; or it may exist independently and not assembled into the electronic device.

[0080] The aforementioned computer-readable medium carries one or more programs, which, when executed by the electronic device, cause the electronic device to perform the method for determining the catalytic efficacy of the aforementioned artificial ice core material.

[0081] Electronic devices can be programmed with computer program code in one or more programming languages ​​or combinations thereof to perform the operations of this disclosure. These programming languages ​​include, but are not limited to, object-oriented programming languages ​​such as Java, Smalltalk, and C++, as well as conventional procedural programming languages ​​such as "C" or similar languages. The program code can be executed entirely on the user's computer, partially on the user's computer, as a standalone software package, partially on the user's computer and partially on a remote computer, or entirely on a remote computer or server. In cases involving remote computers, the remote computer can be connected to the user's computer via any type of network—including a local area network (LAN) or a wide area network (WAN)—or can be connected to an external computer (e.g., via the Internet using an Internet service provider).

[0082] The flowcharts and block diagrams in the accompanying drawings illustrate the architecture, functionality, and operation of possible implementations of systems, methods, and computer program products according to various embodiments of this disclosure. In this regard, each block in a flowchart or block diagram may represent a module, segment, or portion of code containing one or more executable instructions for implementing a specified logical function. It should also be noted that in some alternative implementations, the functions indicated in the blocks may occur in a different order than those indicated in the drawings. For example, two consecutively indicated blocks may actually be executed substantially in parallel, and they may sometimes be executed in reverse order, depending on the functions involved. It should also be noted that each block in the block diagrams and / or flowcharts, and combinations of blocks in the block diagrams and / or flowcharts, can be implemented using a dedicated hardware-based system that performs the specified function or operation, or using a combination of dedicated hardware and computer instructions.

[0083] The units described in the embodiments of this disclosure can be implemented in software or hardware. The names of the units are not, in some cases, intended to limit the specific unit.

[0084] The functions described above in this document can be performed at least in part by one or more hardware logic components. For example, exemplary types of hardware logic components that can be used, without limitation, include: field-programmable gate arrays (FPGAs), application-specific integrated circuits (ASICs), application-specific standard products (ASSPs), system-on-a-chip (SoCs), complex programmable logic devices (CPLDs), and so on.

[0085] In the context of this disclosure, a machine-readable medium can be a tangible medium that may contain or store a program for use by or in conjunction with an instruction execution system, apparatus, or device. A machine-readable medium can be a machine-readable signal medium or a machine-readable storage medium. A machine-readable medium can be, but is not limited to, electronic, magnetic, optical, electromagnetic, infrared, or semiconductor systems, apparatus, or devices, or any suitable combination of the foregoing. More specific examples of machine-readable storage media include electrical connections based on one or more wires, portable computer disks, hard disks, random access memory (RAM), read-only memory (ROM), erasable programmable read-only memory (EPROM or flash memory), optical fiber, portable compact disk read-only memory (CD-ROM), optical storage devices, magnetic storage devices, or any suitable combination of the foregoing.

[0086] The above description is merely a preferred embodiment of this disclosure and an explanation of the technical principles employed. Those skilled in the art should understand that the scope of this disclosure is not limited to technical solutions formed by specific combinations of the above-described technical features, but should also cover other technical solutions formed by arbitrary combinations of the above-described technical features or their equivalents without departing from the above-described concept. For example, technical solutions formed by substituting the above features with (but not limited to) technical features disclosed in this disclosure that have similar functions.

[0087] Furthermore, while the operations are described in a specific order, this should not be construed as requiring these operations to be performed in the specific order shown or in a sequential order. In certain environments, multitasking and parallel processing may be advantageous. Similarly, while several specific implementation details are included in the above discussion, these should not be construed as limiting the scope of this disclosure. Certain features described in the context of individual embodiments may also be implemented in combination in a single embodiment. Conversely, various features described in the context of a single embodiment may also be implemented individually or in any suitable sub-combination in multiple embodiments.

[0088] Although the subject matter has been described using language specific to structural features and / or methodological logic, it should be understood that the subject matter defined in the appended claims is not necessarily limited to the specific features or actions described above. Rather, the specific features and actions described above are merely illustrative examples of implementing the claims.

Claims

1. A method for determining the catalytic efficiency of an artificial ice core material, characterized in that, include: The sample tilt angle and projection sequence of the biomacromolecule multiscale complex are obtained according to the preset first observation conditions. The sample tilt angle and projection sequence are reconstructed and decoded to obtain the three-dimensional reconstruction map and lattice mismatch of the biomacromolecule multiscale complex, wherein the biomacromolecule multiscale complex is embedded in an ice carrier. The observation field of view is determined based on the three-dimensional reconstruction map and the lattice mismatch. The nascent micro ice crystals are observed according to the observation field of view and the preset second observation conditions, and a nucleation spatial probability hotspot map is drawn. The nucleation spatial probability hotspot map is analyzed to obtain the thermodynamic evaluation result of the biomacromolecule multiscale complex. The nascent micro ice crystals are obtained by the biomacromolecule multiscale complex after a preset treatment. The process image of the formation of the new ice crystal front is obtained based on the preset third observation conditions. The advancement contour line of the new ice crystal front is extracted from the process image based on the spatial reference. The differentiated time trajectory path of the new ice crystal front in the preset region is determined according to the nucleation spatial probability hotspot map. The new ice crystal front is obtained by phase transition reaction of biomacromolecule multiscale complex. The spatial reference is obtained by the core nucleation active center identified in the thermodynamic evaluation results. A spatiotemporal map is obtained based on the differentiated time trajectory path and the propulsion contour line. The catalytic rate order-of-magnitude transition of the newly formed ice crystal front is determined based on the spatiotemporal map. Based on the catalytic rate order-of-magnitude transition, the quantitative result of the extreme catalytic efficiency induced by the multi-scale complex of the biomacromolecule to non-classical crystallization is obtained.

2. The method according to claim 1, characterized in that, The sample tilt angle and projection sequence of the biomacromolecule multiscale complex are obtained according to the preset first observation conditions. The sample tilt angle and projection sequence are reconstructed and decoded to obtain the three-dimensional reconstruction map and lattice mismatch of the biomacromolecule multiscale complex. The biomacromolecule multiscale complex is embedded in an ice carrier, including: Based on the environmental and equipment parameters in the preset first observation conditions, a series of tilt imaging was performed on the biomacromolecule multiscale complex using cryo-electron tomography to obtain the sample tilt angle and projection sequence of the biomacromolecule multiscale complex. The common structural features in each projection image of the series of tilt imaging are extracted. The sample tilt angle is aligned with the projection sequence based on the common structural features to obtain the alignment result. The alignment result is then reconstructed by back projection to obtain the three-dimensional electron density map of the biomacromolecule multiscale complex in the ice carrier. The three-dimensional electron density map is decoded to obtain the three-dimensional multipolymer conformation information of the biomacromolecule multiscale complex in the ice carrier, and a three-dimensional reconstruction map is drawn based on the three-dimensional multipolymer conformation information. Based on the three-dimensional reconstructed image, a confined region high-resolution imaging of the phase interface between the biomacromolecule multiscale complex and the ice carrier is performed, local fast Fourier transform is extracted, and the lattice mismatch of the biomacromolecule multiscale complex is calculated.

3. The method according to claim 1, characterized in that, The observation field of view is determined based on the three-dimensional reconstruction image and lattice mismatch. Primary micro-ice crystals are observed according to the observation field of view and preset second observation conditions, and a nucleation spatial probability hotspot map is plotted. The nucleation spatial probability hotspot map is analyzed to obtain the thermodynamic evaluation results of the biomacromolecule multi-scale complex. The primary micro-ice crystals are obtained by pre-processing the biomacromolecule multi-scale complex. The preset processing includes in-situ micro-sublimation and targeted re-sublimation, including: Based on the three-dimensional reconstruction image and lattice mismatch, the observation field of view is determined. Based on the observation field of view and the preset second observation conditions, the ice carrier containing the biomacromolecule multiscale complex is induced to perform in-situ micro-sublimation, exposing the characteristic surface and interface of the biomacromolecule multiscale complex. Based on the characteristic interface and the preset second observation conditions, targeted resublimation is performed on the biomacromolecule multiscale complex to obtain nascent micro ice crystals; Based on the three-dimensional reconstruction map, the nascent micro ice crystals are anchored in three-dimensional space coordinates, and a nucleation space probability hotspot map is drawn. The distribution of the nascent micro ice crystals is determined by analyzing the nucleation space probability hotspot map. Based on the distribution, high-probability aggregation regions of nascent micro ice crystals are extracted from the nucleation space probability hotspot map, and the high-probability aggregation regions are mapped to specific conformational regions of the biomacromolecule multiscale complex. Based on the specific conformational region and the heterogeneous nucleation theory, the microscopic contact angle between the primary micro-ice crystals and the specific conformational region is determined. Based on the microscopic contact angle, the heterogeneous nucleation free energy barrier of the specific conformational region is calculated. The thermodynamic evaluation results of the multi-scale complex of the biomacromolecule are obtained based on the heterogeneous nucleation free energy barrier.

4. The method according to claim 3, characterized in that, The process image of the formation of a new ice crystal front is acquired based on a preset third observation condition. The advancing contour line of the new ice crystal front is extracted from the process image based on a spatial reference. The differentiated time-trajectory path of the new ice crystal front in a preset region is determined according to the nucleation spatial probability hotspot map. The new ice crystal front is obtained by a phase transition reaction of a multi-scale complex of biological macromolecules. The spatial reference is obtained from the core nucleation active centers identified in the thermodynamic evaluation results, including: Based on the preset third observation conditions, pre-cooled water vapor is introduced into the space where the biomacromolecule multiscale complex is located to stimulate the phase transition reaction of the biomacromolecule multiscale complex and obtain the newly formed ice crystal front. The process image of the phase change reaction is obtained, and the process image is analyzed frame by frame to extract the advancing contour line of the newly formed ice crystal front during the phase change reaction process; Using the core nucleation active center identified in the thermodynamic evaluation results as a spatial reference, the differentiated time trajectory path of the newly formed ice crystal front as it slides through a preset region is determined based on the nucleation spatial probability hotspot map. The preset region includes an inactive background bearing area and a specific active region that spans multi-scale complexes.

5. The method according to claim 4, characterized in that, Based on the differentiated time-track path and the propagation contour, a spatiotemporal map is obtained. Based on this map, the order-of-magnitude transition in the catalytic rate of the newly formed ice crystal front is determined. Based on this order-of-magnitude transition in the catalytic rate, the quantitative results of the extreme catalytic efficiency induced by the multi-scale complex of the biomacromolecule to non-classical crystallization are obtained, including: The spatial location information of the newly formed ice crystal front in the differentiated time trajectory path is fused and superimposed with the edge morphology information of the newly formed ice crystal front in the propulsion contour line to obtain a spatiotemporal map. Based on the spatiotemporal map, for each pair of adjacent frame process images, the absolute displacement scale of the newly formed ice crystal front advancing along the normal direction is extracted, along with the time interval corresponding to the absolute displacement scale. The displacement-time series dataset of the phase change reaction process is obtained by continuously extracting the process images frame by frame. The transient dynamic propulsion rate of the newly formed ice crystal front is calculated based on the displacement-time series dataset. Based on the specific active region of the multi-scale complex in the preset region and the transient dynamic propulsion rate, the catalytic rate order-of-magnitude transition of the newly formed ice crystal front is extracted. Using the order-of-magnitude jump in the catalytic rate as a quantitative indicator, the quantitative results of the extreme catalytic efficiency induced by the multi-scale complex of the biomacromolecule to induce non-classical crystallization were obtained.

6. An apparatus for determining the catalytic efficacy of an artificial ice core material, the apparatus comprising: The acquisition module is used to acquire the sample tilt angle and projection sequence of the biomacromolecule multiscale complex according to the preset first observation conditions, reconstruct and decode the sample tilt angle and projection sequence to obtain the three-dimensional reconstruction map and lattice mismatch of the biomacromolecule multiscale complex, wherein the biomacromolecule multiscale complex is embedded in an ice carrier. The analysis module is used to determine the observation field of view based on the three-dimensional reconstruction map and the lattice mismatch, observe the nascent micro ice crystals according to the observation field of view and the preset second observation conditions and draw the nucleation spatial probability hotspot map, analyze the nucleation spatial probability hotspot map, and obtain the thermodynamic evaluation result of the biomacromolecule multiscale complex, wherein the nascent micro ice crystals are obtained by the biomacromolecule multiscale complex under preset processing. The extraction module is used to acquire process images of the formation of new ice crystal fronts based on preset third observation conditions, extract the advancing contour lines of the new ice crystal fronts from the process images based on spatial references, and determine the differentiated time trajectory paths of the new ice crystal fronts in preset regions according to the nucleation spatial probability hotspot map, wherein the new ice crystal fronts are obtained by phase transition reactions of multi-scale complexes of biological macromolecules, and the spatial references are obtained by the core nucleation active centers identified in the thermodynamic evaluation results; The determination module is used to obtain a spatiotemporal map based on the differentiated time trajectory path and the propulsion contour line, determine the order-of-magnitude transition of the catalytic rate of the newly formed ice crystal front based on the spatiotemporal map, and obtain the quantitative result of the extreme catalytic efficiency induced by the multi-scale complex of the biomacromolecule to induce non-classical crystallization based on the order-of-magnitude transition of the catalytic rate.

7. The apparatus according to claim 6, characterized in that, The acquisition module is used for: Based on the environmental and equipment parameters in the preset first observation conditions, a series of tilt imaging was performed on the biomacromolecule multiscale complex using cryo-electron tomography to obtain the sample tilt angle and projection sequence of the biomacromolecule multiscale complex. The common structural features in each projection image of the series of tilt imaging are extracted. The sample tilt angle is aligned with the projection sequence based on the common structural features to obtain the alignment result. The alignment result is then reconstructed by back projection to obtain the three-dimensional electron density map of the biomacromolecule multiscale complex in the ice carrier. The three-dimensional electron density map is decoded to obtain the three-dimensional multipolymer conformation information of the biomacromolecule multiscale complex in the ice carrier, and a three-dimensional reconstruction map is drawn based on the three-dimensional multipolymer conformation information. Based on the three-dimensional reconstructed image, a confined region high-resolution imaging of the phase interface between the biomacromolecule multiscale complex and the ice carrier is performed, local fast Fourier transform is extracted, and the lattice mismatch of the biomacromolecule multiscale complex is calculated.

8. The apparatus according to claim 6, characterized in that, The analysis module is used for: Based on the three-dimensional reconstruction image and lattice mismatch, the observation field of view is determined. Based on the observation field of view and the preset second observation conditions, the ice carrier containing the biomacromolecule multiscale complex is induced to perform in-situ micro-sublimation, exposing the characteristic surface and interface of the biomacromolecule multiscale complex. Based on the characteristic interface and the preset second observation conditions, targeted resublimation is performed on the biomacromolecule multiscale complex to obtain nascent micro ice crystals; Based on the three-dimensional reconstruction map, the nascent micro ice crystals are anchored in three-dimensional space coordinates, and a nucleation space probability hotspot map is drawn. The distribution of the nascent micro ice crystals is determined by analyzing the nucleation space probability hotspot map. Based on the distribution, high-probability aggregation regions of nascent micro ice crystals are extracted from the nucleation space probability hotspot map, and the high-probability aggregation regions are mapped to specific conformational regions of the biomacromolecule multiscale complex. Based on the specific conformational region and the heterogeneous nucleation theory, the microscopic contact angle between the primary micro-ice crystals and the specific conformational region is determined. Based on the microscopic contact angle, the heterogeneous nucleation free energy barrier of the specific conformational region is calculated. The thermodynamic evaluation results of the multi-scale complex of the biomacromolecule are obtained based on the heterogeneous nucleation free energy barrier.

9. The apparatus according to claim 8, characterized in that, The extraction module is used for: Based on the preset third observation conditions, pre-cooled water vapor is introduced into the space where the biomacromolecule multiscale complex is located to stimulate the phase transition reaction of the biomacromolecule multiscale complex and obtain the newly formed ice crystal front. The process image of the phase change reaction is obtained, and the process image is analyzed frame by frame to extract the advancing contour line of the newly formed ice crystal front during the phase change reaction process; Using the core nucleation active center identified in the thermodynamic evaluation results as a spatial reference, the differentiated time trajectory path of the newly formed ice crystal front as it slides through a preset region is determined based on the nucleation spatial probability hotspot map. The preset region includes an inactive background bearing area and a specific active region that spans multi-scale complexes.

10. The apparatus according to claim 9, characterized in that, The determining module is used for: The spatial location information of the newly formed ice crystal front in the differentiated time trajectory path is fused and superimposed with the edge morphology information of the newly formed ice crystal front in the propulsion contour line to obtain a spatiotemporal map. Based on the spatiotemporal map, for each pair of adjacent frame process images, the absolute displacement scale of the newly formed ice crystal front advancing along the normal direction is extracted, along with the time interval corresponding to the absolute displacement scale. The displacement-time series dataset of the phase change reaction process is obtained by continuously extracting the process images frame by frame. The transient dynamic propulsion rate of the newly formed ice crystal front is calculated based on the displacement-time series dataset. Based on the specific active region of the multi-scale complex in the preset region and the transient dynamic propulsion rate, the catalytic rate order-of-magnitude transition of the newly formed ice crystal front is extracted. Using the order-of-magnitude jump in the catalytic rate as a quantitative indicator, the quantitative results of the extreme catalytic efficiency induced by the multi-scale complex of the biomacromolecule to induce non-classical crystallization were obtained.

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