Application of -86 °C low temperature refrigerator on frozen section
With the continuous development of ultra-low temperature refrigerator technology, ultra-low temperature refrigerators are gradually being widely used in the medical field. For example, the application of a -86 ° C low temperature refrigerator on frozen sections is the rapid freezing in the frozen section to reduce the formation of ice crystals.
One of the commonly used slicing methods in immunohistochemical staining is frozen sectioning. Its outstanding advantage is that it can preserve the immunological activity of multiple antigens better, especially the cell surface antigen, and should be frozen. Fresh tissue and fixed tissue can be frozen. When frozen, the water in the tissue is easy to form ice crystals, which often affects the antigen localization. It is generally believed that when the ice crystals are small and large, the influence is small. When the ice crystals are small and long, the damage to the structure is large, and the above phenomenon is more likely to occur in tissues with more water content. The size of the ice crystal is proportional to its growth rate, and inversely proportional to the nucleation rate (forming rate), that is, the more the number of ice crystals formed, the smaller the influence on the tissue structure. Therefore, the number of ice crystals should be reduced as much as possible. Fish believes that the ice nucleation rate is slower at the beginning of freezing, and then gradually increases. The critical temperature is -33 ° C, from -30 ° C to -43 ° C, the nucleation rate increases sharply to 1018, and then slows down. Based on the above theory, the following measures can be taken to reduce the formation of ice crystals.
(1) Quick freezing, which causes the temperature of the tissue to drop rapidly, shortening the time from -33 ° C to -43 ° C, and reducing the formation of ice crystals.
There are two ways:
1 dry ice - acetone (alcohol) method: 150-200ml acetone (alcohol) into a small vacuum flask, gradually added dry ice, until the saturation is viscous, and then add dry ice no longer bubble, the temperature can reach -70 ° C Use a small beaker (50-100ml) containing about 50ml of isopentane, and then slowly place the beaker into the saturated solution of dry ice acetone (pure alcohol) until the isopentane temperature reaches -70 °C. The tissue (size 1 cm×0.8 cm×0.5 cm) was placed in isopentane for 30-60 s and then taken out, or placed in a constant cold box for slicing, or stored in a -86 ° C low temperature refrigerator.
2 liquid nitrogen method: put the tissue block flat in the soft plastic bottle cap or special small box (about 2cm in diameter), such as tissue block small amount can be immersed in the OCT embedding agent, then put the special small box slowly into the flat In the small cup containing liquid nitrogen, when the bottom of the box contacts liquid nitrogen, the gasification begins to boil. At this time, the small box remains in place and must not be immersed in liquid nitrogen. The tissue is rapidly iced into pieces in about 10-20 s. The tissue ice was taken out and placed in a -86 ° C low temperature refrigerator for storage, or placed in a cryostat slicer.
-86 °C low temperature refrigerator parameters:
Product Series | model | Volume L | Temperature °C | Inner size: width × depth × height mm | Outer size: width × depth × height mm | Power W | Net weight Kg |
-86 ° C | TF-86-30-LA | 30 | -25~-65 | 270x265x400 | 470x490x1120 | 200 | 60 |
TF-86-60-LA | 60 | -25~-65 | 364x405x409 | 540x620x1115 | 240 | 75 | |
TF-86-80-LA | 80 | -40~-86 | 380×410×500 | 620×660×1275 | 450 | 80 | |
TF-86-158-LA | 158 | -40~-86 | 380×415×1020 | 620×660×1795 | 500 | 100 | |
TF-86-328-LA | 328 | -40~-86 | 500×500×1300 | 760×765×1975 | 550 | 170 | |
TF-86-400-LA | 400 | -40~-86 | 500×600×1300 | 760×865×1975 | 650 | 190 | |
TF-86-550-LA | 550 | -40~-86 | 600×700×1300 | 860×965×1975 | 800 | 220 | |
TF-86-638-LA | 638 | -40~-86 | 700×700×1300 | 960×965×1975 | 1000 | 250 | |
TF-86-750-LA | 750 | -40~-86 | 800×700×1300 | 1060×965×1975 | 1200 | 280 | |
TF-86-938-LA | 938 | -40~-86 | 1000×700×1300 | 1260×965×1975 | 1700 | 320 | |
-86 ° C | TF-86-50-WA | 50 | -40~-86 | 440×350×315 | 650×560×880 | 350 | 60 |
TF-86-105-WA | 105 | -40~-86 | 410×460×550 | 1140×760×930 | 400 | 105 | |
TF-86-200-WA | 200 | -40~-86 | 710×510×560 | 1390×760×930 | 500 | 160 | |
TF-86-300-WA | 300 | -40~-86 | 1010×510×580 | 1690×760×930 | 600 | 180 | |
TF-86-480-WA | 480 | -40~-86 | 1300×600×600 | 1990×860×975 | 700 | 240 |
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