Freezing method
A freezing method and a technology of magnetic field frequency, applied in refrigerators, preservation of meat/fish through freezing/cooling, refrigeration and liquefaction, etc., can solve the problems of not understanding the relationship between magnetic field frequency and supercooling state, and not fully understanding the relationship, etc., to achieve The effect of high supercooling
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example 1
[0044] A coil with an inner diameter of 80 mm and a depth of 80 mm (6 turns) is arranged outside an aluminum cooling tank with an inner diameter of 40 mm and a depth of 40 mm, and the coil and the cooling tank are installed in a cooling device (Stirling engine, Twinbird Company, SC-UD08) superior. Connect a function generator for generating an alternating current to the coil. A freezing experiment was carried out by cooling 1 ml of physiological saline (freezing point: -0.65° C.) using this apparatus. Set the magnetic field strength to 0.12±0.02mT, the cooling rate to 0.8K / min, and set the frequency of the AC current to 0Hz (no magnetic field), 50Hz, 200Hz, 2000Hz, 20000Hz or 200000Hz, 12 times for different magnetic field frequencies experiment. figure 1 The temperature change after performing 3 tests in the case of a magnetic field frequency of 50 Hz is shown. For each test, the "degree of supercooling" is determined as the absolute value of the difference between the fre...
example 2
[0047] After starving for 24 hours, 2 Lewis female mice (285g, 280g) were euthanized under ether anesthesia, and then directly used for the freezing experiment without removing the organs. Fill the cooling tank of the freezer with 60% ethylene glycol, remove air as much as possible, put the mouse sealed in the nylon bag into the measuring instrument with a sinker, and sink into the cooling tank. The magnetic field strength was set at 0.1 mT to 0.2 mT, and after cooling at 2° C. for 1 hour, cooling was performed at a rate of -0.5° C. / min, and cooling was carried out to -30° C. for freezing. It took 2 hours and 45 minutes from the start of cooling to the end of freezing. The frequency of the magnetic field is 2000Hz.
[0048] Tissue sections were cut from the frozen tissue according to the thickness of 5-7 μm, and the sections were pasted on glass slides and air-dried for 30 minutes. Staining of tissue sections was performed by the following method using hematoxylin / eosin stai...
example 3
[0051] Using alkaline phosphatase (ALP) and green fluorescent protein (GFP), freezing and thawing by the method of the present invention were repeated to confirm the degree of modification. Put 100 μl of sample into a sample tube made of polyethylene, store it in a stainless steel tube (1×1×4cm) filled with aluminum balls (diameter 0.5mm), and store it under a magnetic field frequency of 30Hz or 2000Hz and a magnetic field strength of 0.8mT Under the conditions, freezing conditions: 45 minutes from 30°C to -40°C and thawing conditions: 20 minutes after freezing to 30°C were set as one cycle, and freezing / thawing was repeated 6 times.
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