Hollow refrigeration device for observing ice crystal growth process under optical microscope

Abstract

The utility model provides a hollow refrigeration device for observing the growth process of ice crystals under an optical microscope, which is characterized in that a hollow water-cooling head, a hollow semiconductor chilling plate and a glass plate are sequentially fixed on an objective table of the microscope, and the water-cooling head is communicated with an external circulating water pump, a heat preservation tank and a refrigerator through a circulating pipeline; an adjustable power supply is adopted to independently control the semiconductor chilling plate, and the circulating water pump is connected with the refrigerator. In a limited space between the objective table and the objective lens of the glass microscope, the hollow semiconductor chilling plate and the water cooling head are adopted to form a compact refrigeration table, and then the compact refrigeration table is connected to an external circulating medium through the hose, so that the cooling efficiency and the control precision are remarkably improved, and the ice crystal morphology at different crystallization moments can be observed in real time.

Description

Technical Field

[0001] The utility model relates to a hollow refrigeration device used for observing the growth process of ice crystals under an optical microscope, and is used for observing and studying the growth process of ice crystals of antifreeze protein solutions in laboratories. Background Art

[0002] In low-temperature environments, the presence of water causes a series of physical and chemical changes in substances. In organisms, freezing of aqueous solutions can cause cell death and affect some functions of the organism. Therefore, organisms have evolved a series of cold resistance mechanisms, and antifreeze protein is one of them. Antifreeze protein is a substance that can non-colligatively lower the freezing point in low-temperature environments, change the morphology of ice crystals, and inhibit ice recrystallization to ensure the activity of organisms.

[0003] At present, the observation and research of antifreeze proteins are mainly carried out using cryogenic microscopes or microscopes combined with cryogenic stages. Cryogenic microscopes perform electron microscopic imaging of samples under liquid nitrogen / helium conditions, and there are many supporting facilities. Some cryogenic stages are opaque, some are translucent but have poor refrigeration effects, and some are translucent and have good refrigeration effects, but have complex structures and high costs. At the same time, the observation effects of these cold stages are uneven and cannot be promoted on a large scale. Therefore, how to meet the requirements of light transmission while refrigerating has become a key technology for observing the growth morphology of ice crystals. The observation of the growth morphology of ice crystals requires the cold stage to have high cooling efficiency, stable temperature control, and good light transmittance to reflect the antifreeze effect of antifreeze proteins on water.

[0004] In view of the fact that the current light transmittance, cooling efficiency and structure of the cooling platform cannot meet the research needs of most scholars, it is proposed to use semiconductor cooling sheets in combination with water cooling heads to solve the above problems. The semiconductor cooling sheet uses the Peltier effect of semiconductor materials. When direct current passes through a couple formed by two different semiconductor materials in series, heat can be absorbed and released at both ends of the couple, achieving the purpose of cooling. It has the characteristics of light weight, small size, low noise and high cooling efficiency.

[0005] After searching, Chinese patent document CN201615966U discloses a refrigerated stage for optical microscopes, which has a relatively complex structure and operation. CN204666947U discloses a microscope ice crystal cold stage, which solves the problem of ice crystal samples melting easily when observed under a microscope in experiments through semiconductor refrigeration and groove design, and reduces the experimental error caused by the melting of ice crystal samples exposed to the environment. These refrigeration devices have a narrow temperature control range and a single function, and cannot meet the requirements of experimental observations for cooling efficiency and stable temperature control. Summary of the invention

[0006] Purpose of the invention: In order to overcome the deficiencies in the prior art, the present invention designs a hollow refrigeration device for observing the growth process of ice crystals under an optical microscope.

[0007] Technical solution: To solve the above technical problems, the present invention provides a hollow refrigeration device for observing the growth process of ice crystals under an optical microscope, comprising a semiconductor refrigeration sheet, a water cooling head, an insulation tank, a refrigerator, a power supply, a circulating water pump, a circulating water pipe, a glass plate and an acrylic box;

[0008] The glass plate is placed on a semiconductor refrigeration sheet for placing samples; the semiconductor refrigeration sheet is adhered to a water cooling head by means of thermal grease; the water cooling head fixes the refrigeration system on a microscope stage by means of a sheet clamp; the water cooling head is provided with water inlet and outlet channels; the inlet of the channel is connected to the outlet of a circulating water pump; the circulating water pump is placed in an insulation tank; the outlet of the channel is connected to the inlet of a refrigerator; the outlet of the refrigerator is connected to an insulation tank; the insulation tank and the refrigerator are placed in an acrylic box; the power supply includes a first power supply and a second power supply, the first power supply is connected to the semiconductor refrigeration sheet, and the second power supply is connected to the circulating water pump and the refrigerator.

[0009] Furthermore, the semiconductor cooling plate is annular and hollow, which is convenient for the light source of the optical microscope to pass through. That is, a through hole for the light source to pass through is formed in the center of the semiconductor cooling plate.

[0010] Furthermore, the water cooling head is made of aluminum and is hollow, and is placed under the semiconductor refrigeration plate to facilitate cooling the semiconductor refrigeration plate.

[0011] Furthermore, the circulating water pipe is a hose, which is convenient for assembly with various components.

[0012] Furthermore, the circulating water pipe is fixed to each component by a stainless steel hose clamp to prevent leakage of the circulating liquid.

[0013] Furthermore, the refrigerator is composed of a fan and an annular circuitous channel, which is convenient for rapid cooling.

[0014] Furthermore, the first power supply is a voltage-adjustable power supply, and the voltage adjustment knob of the adjustable power supply is placed on one side of the microscope, which is used to independently control the freezing rate during the observation of ice crystal growth and melting. The second power supply is an adjustable power supply, which is used to independently control the circulation speed of the circulating fluid and the refrigeration effect. The operator needs to use both hands and eyes to operate the adjustable power supply while observing the eyepiece of the microscope. First, adjust the voltage to 5V, increase or decrease the voltage according to the dynamic process of the freezing situation, and after the solution is completely frozen, modulate the voltage to 12V to ensure the refrigeration effect; after observing the morphology of the ice crystals, lower the voltage to 3V, and the solution slowly dissolves. According to the dynamic process of the melting situation, increase or decrease the power supply voltage to observe the melting process of the ice crystals.

[0015] Furthermore, the circulating liquid is a liquid with good thermal conductivity, such as ethylene glycol solution. The liquid is contained in an insulation tank. When the power is turned on, the circulating water pump and the refrigerator start to operate, driving the circulating liquid in the insulation tank to flow to the refrigerator. The refrigeration system in the refrigerator further cools the circulating liquid. After refrigeration, the circulating liquid flows to the hollow water cooling head. After the hollow semiconductor refrigeration plate is connected to the power supply, it cools on one side for cooling the sample and generates heat on the other side. The heat generated is taken away by the circulating liquid of the hollow water cooling head. After flowing through the hollow water cooling head, the circulating liquid returns to the insulation tank, completing a cycle, which is convenient for improving the refrigeration efficiency.

[0016] The beneficial effects of this patent are: by arranging a semiconductor refrigeration sheet under the glass plate, the object to be measured placed on the glass plate can be cooled, and at the same time, cooling liquid is passed into the water-cooling head under the semiconductor refrigeration sheet to take away the heat generated by the semiconductor refrigeration sheet and improve the refrigeration efficiency. The hollow and light-transmitting design allows this patented device to observe the process of liquid crystallization under an optical microscope, and the morphology of ice crystals at different crystallization moments can be observed in real time by adjusting the microscope magnification.

[0017] The utility model sequentially fixes a hollow water cooling head, a hollow semiconductor cooling sheet and a glass plate on a microscope stage, the water cooling head is connected to an external circulating water pump, a heat preservation tank and a refrigerator through a circulation pipeline, and an adjustable power supply is used to independently control the semiconductor cooling sheet, the circulating water pump and the refrigerator. In the limited space between the glass microscope stage and the objective lens, a hollow semiconductor cooling sheet and a water cooling head are used to form an ultra-thin miniaturized cooling stage, which is then connected to an external circulating medium with a hose, so that the cooling efficiency and control accuracy are significantly improved. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 It is a schematic diagram of the structure of Embodiment 1 of the present utility model;

[0019] Figure 2 for Figure 1 A schematic diagram of the structure of the internal channel of the middle refrigerator;

[0020] Figure 3 for Figure 1 Schematic diagram of the cooling part under the microscope;

[0021] Figure 4 yes Figure 1 Dimensional diagram of the hollow water cooling head;

[0022] Figure 5 is the morphology of ice crystals in aqueous solution (40X);

[0023] Figure 6 is the morphology of ice crystals in antifreeze protein solution (40X);

[0024] In the figure, there are a heat preservation tank 1, a circulating water pump 2, a refrigerator 3, a power adapter 4, an acrylic box 5, an internal structure of the refrigerator 6, a circulating water pipe 7, a power adapter 8, a ring-shaped hollow semiconductor refrigeration sheet 9, a hollow water cooling head 10, and a glass plate 11. DETAILED DESCRIPTION

[0025] Embodiment 1:

[0026] The purpose and advantages of this patent are further explained below in conjunction with the accompanying drawings.

[0027] like Figure 1-2 As shown, the hollow refrigeration device for observing the growth process of ice crystals under an optical microscope of this embodiment includes an insulation tank 1, a circulating water pump 2, a refrigerator 3, a power adapter 4, an acrylic box 5, a circulating water pipe 7, a power adapter 8, an annular hollow semiconductor refrigeration sheet 9 and a hollow water cooling head 10; wherein, the glass plate is placed above the annular hollow semiconductor refrigeration sheet 9 for placing samples; the annular hollow semiconductor refrigeration sheet 9 is connected to the hollow water cooling head 10 through thermal conductive silicone grease to facilitate taking away the heat generated by the semiconductor refrigeration sheet 9; the annular hollow semiconductor refrigeration sheet 9 and the hollow water cooling head 10 It is fixed on the stage of the microscope by a sheet clamp; the hollow water cooling head 10 is provided with water inlet and outlet channels respectively; the inlet of the channel is connected to the outlet of the circulating water pump 2 in the heat preservation tank 1; the outlet of the channel is connected to the inlet 6 of the refrigerator 3; the refrigerator 3 has a circular circuitous pipeline inside, and a fan is arranged behind the pipeline to cool the circulating fluid; the outlet of the refrigerator 3 is connected to the heat preservation tank 1; the heat preservation tank 1 and the refrigerator 3 are placed in an acrylic box 5; the power adapter 4 is connected to the circulating water pump 2 and the refrigerator 3, and the power adapter 8 is connected to the annular hollow semiconductor cooling sheet 9. The heat preservation tank contains an ethylene glycol solution at -20℃ to -18℃, and the circulating water pipe 7 is a hose. The cooling surface of the annular hollow semiconductor cooling sheet 9 faces upward, and the heat dissipation surface faces downward, and thermal conductive silicone grease is applied between the annular hollow semiconductor cooling sheet 9 and the hollow water cooling head 10.

[0028] The semiconductor cooling chip adopts the TEC1-1206 semiconductor cooling chip produced by Jiangsu Xinghe Electronics Co., Ltd., and its appearance size is 40mm×40mm×3.95mm; according to the specifications of the semiconductor cooling chip, it is customized by Hualian Metal Technology Co., Ltd. as shown in the attached Figure 1 The hollow water cooling head of the size shown adopts an insulation tank with a diameter of 8cm and a height of 9cm produced by Yuliang Electronics Co., Ltd., a cooler with a 120mm / 18 straight pipe mouth, a 12V5W DC brushless silent water pump, and a PVC pipe with an inner diameter of 8mm and an outer diameter of 10mm.

[0029] Embodiment 2:

[0030] (1) Preparation of antifreeze protein solution: Weigh 500 g of antifreeze protein powder using a 1 / 1000 balance, put it into a beaker, add 50 mL of ultrapure water to dissolve it, and then put it into a 100 mL volumetric flask. Then pour 20 mL of ultrapure water into the beaker to wash off the antifreeze protein remaining on the beaker. Then transfer the solution to a 100 mL volumetric flask. Finally, pour 20 mL of ultrapure water to wash off the antifreeze protein remaining on the wall of the beaker. Add ultrapure water to the volumetric flask and make up to the 100 mL mark.

[0031] (2) Preparation of refrigeration device: Place the circulating water pump 2 into the insulation tube 1, connect the circulating water pump and the water inlet of the hollow water cooling head 10 through the circulating water pipe 7, connect the water outlet of the hollow water cooling head 10 to the water inlet of the refrigerator 3 through the circulating water pipe 7, connect the water outlet of the refrigerator to the insulation tube through the circulating water pipe 7, stick the hollow semiconductor cooling sheet 9 to the hollow water cooling head 10 with thermal conductive silicone grease, connect the hollow semiconductor cooling sheet 9, the circulating water pump 2 and the refrigerator 3 to the power adapter 4 and the power adapter 8, inject ethylene glycol at a temperature of -19±1°C into the insulation tank 1, and place a glass sheet with a thickness of 2 mm on the hollow semiconductor cooling sheet 9.

[0032] (3) Observation of ice crystal morphology: Turn on the power adapter 4 to allow ethylene glycol to circulate in the hollow water cooling head 1, the refrigerator 3 and the insulation tube 1. Use a pipette to draw 10 μL of an antifreeze protein solution with a concentration of 15 mg / mL and drop it evenly on the glass plate. Then turn on the power adapter 8 and adjust the voltage to an appropriate value. Adjust the magnification of the optical microscope until the solution can be clearly observed. As the semiconductor refrigeration sheet 9 operates, the process of the solution slowly freezing can be observed. Further adjust the magnification to observe the morphology of ice crystals. Figure 5 and Figure 6 As shown. The optical microscope can be directly connected to the computer with a display screen and a video recording function.

[0033] Adjusting the power supply voltage is convenient for controlling the freezing rate of the solution and the refrigeration effect, so as to observe the growth and melting process of ice crystals. The specific adjustment method is as follows: first turn on the power supply and adjust the voltage to 5V. According to the dynamic process of the freezing situation, increase or decrease the voltage. After the solution is completely frozen, modulate the voltage to 12V to ensure the refrigeration effect; after observing the morphology of the ice crystals, lower the voltage to 3V, and the solution slowly dissolves. According to the dynamic process of the melting situation, increase or decrease the power supply voltage to observe the melting process of the ice crystals.

Claims

1. A hollow refrigeration device for observing the growth process of ice crystals under an optical microscope, characterized in that: Including semiconductor cooling sheet, water cooling head, insulation tank, refrigerator, power supply, circulating water pump, circulating water pipe, glass plate and acrylic box; The glass plate is placed on a semiconductor refrigeration sheet for placing samples; the semiconductor refrigeration sheet is adhered to a water cooling head by means of thermal grease; the water cooling head fixes the refrigeration system on a microscope stage by means of a sheet clamp; the water cooling head is provided with water inlet and outlet channels; the inlet of the channel is connected to the outlet of a circulating water pump; the circulating water pump is placed in an insulation tank; the outlet of the channel is connected to the inlet of a refrigerator; the outlet of the refrigerator is connected to an insulation tank; the insulation tank and the refrigerator are placed in an acrylic box; the power supply includes a first power supply and a second power supply, the first power supply is connected to the semiconductor refrigeration sheet, and the second power supply is connected to the circulating water pump and the refrigerator.

2. The hollow refrigeration device for observing the ice crystal growth process under an optical microscope according to claim 1, characterized in that: The semiconductor refrigeration plate is annular in shape, and has a through hole in the center for passing the light source.

3. The hollow refrigeration device for observing the ice crystal growth process under an optical microscope according to claim 2, characterized in that: The cooling surface of the semiconductor refrigeration plate faces upwards, and the heat dissipation surface faces downwards; the water cooling head is annular and hollow, and is attached below the semiconductor refrigeration plate.

4. The hollow refrigeration device for observing the ice crystal growth process under an optical microscope according to claim 1, characterized in that: The first power supply is a voltage-adjustable power supply, and the voltage-adjustable power supply's voltage-adjustable knob is placed on one side of the microscope, and is used to control the freezing rate during the observation of ice crystal growth and melting.

5. The hollow refrigeration device for observing the growth process of ice crystals under an optical microscope according to claim 1, characterized in that: The second power supply is an adjustable power supply, which is used to independently control the circulation speed of the circulating fluid.

6. The hollow refrigeration device for observing the growth process of ice crystals under an optical microscope according to claim 1, characterized in that: The circulating water pipe is a hose, and the hose is fixed to each component through a stainless steel hose clamp.

7. The hollow refrigeration device for observing the growth process of ice crystals under an optical microscope according to claim 1, characterized in that: The refrigerator is composed of a fan and an annular circuitous channel.

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