Microscopic camera device for cell freezing and rewarming process

Abstract

The utility model belongs to the technical field of biology, and particularly relates to a microscopic camera device for a cell freezing and rewarming process. Comprising a jet cavity head, a jet outflow pipeline, a profound hypothermia liquid jet pump, a jet pump inlet pipeline, a profound hypothermia liquid tank, a sample cold platform, a cover plate with an observation window, a steam closed cavity, a jet cavity head bracket, an air exhaust interface and a thermocouple. According to the device disclosed by the invention, rapid freezing of cells is realized; in addition, the device has process visual shooting conditions and real-time temperature measurement capability, and can be used for synchronously observing and shooting liquid state, vitrification and crystallization conversion detail processes in the cell freezing process within dozens of milliseconds to several seconds. The method plays an important role in the field of research on cryogenic cell cryopreservation.

Description

Technical Field

[0001] The utility model relates to the field of biotechnology, and in particular to a microscopic imaging device for the process of cell freezing and thawing. Background Technique

[0002] Rapid cell freezing is one of the research hotspots in the biological field, and vitrification freezing is a rapid freezing technology. In order to enable the sample to be in spaced contact with the low-temperature cold surface but be rapidly cooled, and a system for visualizing the rapid freezing process, it is of great value to design and develop a microscopic imaging device for the process of cell freezing and thawing.

[0003] The papers and patents on microscopic imaging systems for observing the rapid vitrification freezing and thawing process of cells that have been publicly disclosed so far have conduction-cooling freezing and temperature-lowering modules that are different from this device. For example, Chinese Patent CN 110057821 A discloses a method for cryopreserving human gamete cells, in particular a low-temperature microscopic imaging system for observing the rapid freezing and thawing process of human gametes. Its conduction-cooling freezing and temperature-lowering module moves the gamete visual environment chamber downward and makes its surface closely contact the liquid nitrogen heat sink cavity. When the movement stops, the liquid nitrogen heat sink cavity quickly transfers the cold to the sample in the gamete visual environment chamber. For example, Chinese Patent CN109997841 B discloses a rapid freezing and thawing visualization environment chamber for human gametes that maintains an atmospheric air atmosphere. Its conduction-cooling freezing and temperature-lowering module moves the entire position of the environment chamber directly above the low-temperature cold surface, and then moves downward until the high-thermal conductivity bottom plate at the bottom of the environment chamber body closely contacts the low-temperature cold surface for rapid temperature reduction. For example, Chinese Patent CN114544317A discloses a freeze-thaw instrument for cell detection. Its conduction-cooling freezing and temperature-lowering module installs the refrigerator inside the thermos flask, and the refrigerating end of the refrigerator is connected to the copper rod. The temperature of the copper rod is reduced by the refrigerator, and the copper rod transfers the temperature to the sample chamber to reduce the temperature of the sample chamber to achieve the purpose of freezing. Content of the Utility Model

[0004] The purpose of the utility model is to provide a microscopic imaging device for the process of cell freezing and thawing, which solves the existing problems.

[0005] In order to achieve the above purpose, the utility model adopts the following technical scheme:

[0006] A microscopic imaging device for the process of cell freezing and thawing. It is characterized in that it includes a spray chamber head, a spray outflow pipe, a cryogenic liquid injection pump, an injection pump inlet pipe, a cryogenic liquid tank, a sample cold platform, a cover plate with an observation window, a steam tight cavity, a spray chamber head bracket, an air extraction interface, and a thermocouple, which together form a conduction-cooling freezing and temperature-lowering module.

[0007] The inlet of the injection cavity head, which is adiabatically installed above the injection cavity head bracket, is connected to the outlet of the cryogenic liquid injection pump through the injection outflow pipe. The inlet of the cryogenic liquid injection pump is connected to the cryogenic liquid tank through the injection pump inlet pipe and extends to the bottom of the cryogenic liquid tank. The sample cold platform is installed below the position of the open hole in the middle of the upper cover plate of the steam-tight cavity; the cover plate with an observation window covers the position of the open hole in the middle of the upper cover plate of the steam-tight cavity. The air extraction interface is connected to the exhaust pipeline. The thermocouple is embedded inside the sample cold platform near the central position.

[0008] The injection cavity head is wrapped with adiabatic thermal insulation cotton, has an optical path through hole in the center, has a fluid pipeline inside, and has injection holes on it.

[0009] The injection flow rate of the cryogenic liquid injection pump is controlled; when it needs to work, the injection flow rate is controlled by comparing and calculating the temperature measured by the thermocouple with the set target temperature. The injection outflow pipe and the injection pump inlet pipe are externally provided with adiabatic thermal insulation measures.

[0010] The central part of the cover plate with an observation window is embedded with highly transparent quartz glass, there is a groove on the lower periphery, and an O-shaped highly adiabatic sealing gasket is embedded in the groove.

[0011] The sample cold platform is made of pure copper or pure silver; it has an optical path through hole in the center and a groove for placing a glass slide on it. The thermocouple is embedded inside the sample cold platform near the central position. When it needs to work, the cryogenic liquid injection pump sprays cryogenic liquid onto the lower surface of the sample cold platform to quickly cool it and conduct the heat to the cell sample on the glass slide.

[0012] The steam-tight cavity is connected to the exhaust pipeline through the air extraction interface; the cryogenic liquid fluid steam generated during operation is discharged through the exhaust pipeline.

[0013] When the cryogenic liquid fluid injection and cooling module of the present utility model is in use, the working process is as follows:

[0014] Placing the sample: Open the cover plate with an observation window, place the glass slide with the cell sample dropped on it in the groove of the sample cold platform; cover the cover plate with an observation window.

[0015] Starting freezing: The cryogenic liquid injection pump sprays cryogenic liquid onto the lower surface of the sample cold platform and conducts the heat to the cell sample on the glass slide to quickly cool it. The injection time and flow rate are controlled by comparing and calculating the temperature measured by the thermocouple with the set target temperature.

[0016] Compared with the prior art, the beneficial effects of the present utility model are as follows:

[0017] 1. The present invention utilizes a cryogenic liquid jetting freezing and cooling module. The cryogenic liquid jet pump sprays the cryogenic liquid onto the lower surface of the sample cooling platform, where it is then conducted to rapidly cool the cell sample on the slide. The sample cooling platform is made of pure copper or silver, offering extremely high cooling efficiency and speed, enabling the cell sample to be rapidly cooled to the evaporation temperature of the cryogenic liquid (e.g., liquid nitrogen -196°C).

[0018] 2. In the present invention, the freezing and cooling module based on the injection of cryogenic liquid fluid, the injection time and flow rate of the cryogenic liquid are calculated and controlled based on the comparison between the temperature measured by the thermocouple and the set target temperature, thereby achieving precise control of the freezing temperature.

[0019] 3. The present invention's refrigeration and cooling module based on metal braided belt conduction cooling generates cryogenic liquid fluid vapor during operation and is discharged through an exhaust pipe, avoiding the observation light path and preventing fogging from affecting observation. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following briefly introduces the drawings required for describing the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work.

[0021] Figure 1 This is a schematic diagram of the three-dimensional structure of a microscopic camera device for cell freezing and thawing process proposed by the present invention;

[0022] Figure 2 This is a partial three-dimensional structural diagram of a microscopic imaging device for cell freezing and thawing process proposed by the present invention;

[0023] In the figure: 1. Injection cavity head; 2. Injection outflow pipe; 3. Cryogenic liquid injection pump; 4. Inlet pipe of injection pump; 5. Cryogenic liquid tank; 6. Sample cooling platform; 7. Cover with observation window; 8. Steam-sealed cavity; 9. Injection cavity head bracket; 10. Vacuum interface; 11. Microscope; 12. High-speed camera; 13. Main bracket; 14. Base plate; 15. Thermocouple. DETAILED DESCRIPTION

[0024] The following will be combined with the accompanying drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0025] Reference Figure 1-2 , please refer to Figures 1 to 2 , as can be seen from the figure, a microscopic imaging device for the cell freezing and thawing process. It includes: 1. Injection chamber head; 2. Injection outflow pipe; 3. Cryogenic liquid injection pump; 4. Injection pump inlet pipe; 5. Cryogenic liquid tank; 6. Sample cold platform; 7. Cover plate with observation window; 8. Vapor tight cavity; 9. Injection chamber head support; 10. Exhaust interface; 15. Thermocouple together form a conduction cooling module for freezing and cooling down.

[0026] The injection chamber head inlet adiabatically installed on the injection chamber head support is connected to the outlet of the cryogenic liquid injection pump through the injection outflow pipe. The inlet of the cryogenic liquid injection pump is connected to the cryogenic liquid tank through the injection pump inlet pipe and extends to the bottom of the cryogenic liquid tank. The sample cold platform is installed below the position of the through hole in the middle of the upper cover plate of the vapor tight cavity; the cover plate with the observation window covers the position of the through hole in the middle of the upper cover plate of the vapor tight cavity. The exhaust interface is connected to the exhaust pipeline. The thermocouple is embedded inside the sample cold platform near the center position.

[0027] The injection chamber head is wrapped with adiabatic insulation cotton, has an optical path through hole in the center, a fluid pipeline inside, and injection holes on it.

[0028] The injection flow rate of the cryogenic liquid injection pump is controlled; when working, the injection flow rate is controlled by comparing and calculating the temperature measured by the thermocouple with the set target temperature. The injection outflow pipe and the injection pump inlet pipe are externally provided with adiabatic insulation measures.

[0029] The center of the cover plate with the observation window is embedded with highly transparent quartz glass, there is a groove on the lower periphery, and an O-shaped highly adiabatic sealing gasket is embedded in the groove.

[0030] The sample cold platform is made of pure copper or pure silver; there is an optical path through hole in the center and a slide placement groove on it. The thermocouple is embedded inside the sample cold platform near the center position. When working, the cryogenic liquid injection pump sprays cryogenic liquid onto the lower surface of the sample cold platform to quickly cool and conduct to the cell sample on the slide.

[0031] The vapor tight cavity is connected to the exhaust pipeline through the exhaust interface; the cryogenic liquid vapor generated during operation is discharged through the exhaust pipeline.

[0032] The implementation principle of a microscopic imaging device for the cell freezing and thawing process in the embodiment of this application is:

[0033] When the cryogenic liquid injection and freezing and cooling module of the present invention is used, the working process is:

[0034] Placing the sample: Open the cover plate with an observation window, place the glass slide with the cell sample dropped thereon in the groove of the sample cold platform; cover the cover plate with the observation window.

[0035] Starting freezing: The cryogenic liquid injection pump sprays cryogenic liquid onto the lower surface of the sample cold platform and conducts it to the cell sample on the glass slide for rapid cooling. The injection time and flow rate are controlled by comparing the temperature measured by the thermocouple with the set target temperature.

[0036] Observing and recording: The microscope consists of 11, a microscope; 12, a high-speed camera; 13, a main bracket; 14, a bottom plate. The observing and imaging system can synchronously observe and record the details of the liquid state, vitrification, and crystallization conversion processes during the cell freezing process within dozens of milliseconds to several seconds.

[0037] In the description of this specification, the descriptions referring to terms such as "one embodiment", "example", "specific example", etc. mean that the specific features, structures, materials, or characteristics described in connection with the embodiment or example are included in at least one embodiment or example of the present utility model. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials, or characteristics described can be combined in a suitable manner in any one or more embodiments or examples.

[0038] The above has introduced in detail a microscopic imaging device for the cell freezing and rewarming process provided by the present utility model. Specific embodiments are used herein to elaborate on the principle and implementation manner of the present utility model. The description of the above embodiments is only used to help understand the method and its core idea of the present utility model. It should be noted that for those of ordinary skill in the art in this technical field, without departing from the principle of the present utility model, several improvements and modifications can still be made to the present utility model, and these improvements and modifications also fall within the protection scope of the claims of the present utility model.

Claims

1. A microscopic imaging device for the cell cryopreservation and rewarming process; characterized in that: It consists of a spray chamber head (1), a spray outflow pipe (2), a cryogenic liquid injection pump (3), an injection pump inlet pipe (4), a cryogenic liquid tank (5), a sample cold platform (6), a cover plate with an observation window (7), a steam tight cavity (8), a spray chamber head bracket (9), an air extraction interface (10), and a thermocouple (15) to jointly form a conduction cooling module. The inlet of the spray chamber head (1) adiabatically installed on the spray chamber head bracket (9) is connected to the outlet of the cryogenic liquid injection pump (3) through the spray outflow pipe (2). The inlet of the cryogenic liquid injection pump (3) is connected to the cryogenic liquid tank (5) through the injection pump inlet pipe (4) and extends to the bottom of the cryogenic liquid tank (5). The sample cold platform is installed below the position of the through hole in the middle of the upper cover plate of the steam tight cavity (8). The cover plate with an observation window (7) covers the position of the through hole in the middle of the upper cover plate of the steam tight cavity (8). The air extraction interface (10) is connected to the exhaust pipeline. The thermocouple (15) is embedded inside the sample cold platform (6) near the center position.

2. The microscopic imaging device for the cell freezing and rewarming process according to claim 1, wherein: The spray chamber head (1) is wrapped with adiabatic thermal insulation cotton, has an optical path through hole in the center, a fluid pipeline inside, and spray holes on it.

3. The microscopic imaging device for the cell freezing and rewarming process according to claim 1, wherein: The injection flow rate of the cryogenic liquid injection pump (3) is controlled. When it needs to work, the injection flow rate is controlled by comparing and calculating the temperature measured by the thermocouple (15) with the set target temperature. The spray outflow pipe (2) and the injection pump inlet pipe (4) are externally provided with adiabatic thermal insulation measures.

4. The microscopic imaging device for the cell freezing and rewarming process according to claim 1, wherein: The center of the cover plate with an observation window (7) is embedded with high-transparency quartz glass, and there is a groove on the lower periphery, and an O-shaped high-adiabatic sealing gasket is embedded in the groove.

5. The microscopic imaging device for the cell freezing and rewarming process according to claim 1, characterized in that: The sample cold platform (6) is made of pure copper or pure silver; it has an optical path through hole in the center and a slide placement groove on it. When it needs to work, the cryogenic liquid injection pump (3) sprays cryogenic liquid onto the lower surface of the sample cold platform (6) to quickly cool and conduct to the cell sample on the slide.

6. The microscopic imaging device for the cell freezing and rewarming process according to claim 1, wherein: The steam tight cavity (8) is connected to the exhaust pipeline through the air extraction interface (10). The cryogenic liquid fluid steam generated during operation is discharged through the exhaust pipeline.

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