OCT冰冻切片包埋剂是一种聚乙二醇和聚乙烯醇的水溶性混合物，目前已广泛用于免疫组化实验中，其用途是在冰冻切片时支撑组织，以增加组织的连续性，减少皱折及碎裂。又因包埋液为水溶性，故在漂片时可溶于水，所以在以后的染色中不会增加背景染色。该试剂仅用于科研领域，不适用于临床诊断或其他用途。
| 项目号 | 颜色 |
F1508750-01 | 无色 |
F1508750-02 | 红色 |
F1508750-03 | 橙色 |
F1508750-04 | 黄色 |
F1508750-05 | 绿色 |
F1508750-06 | 青色 |
F1508750-07 | 蓝色 |
F1508750-08 | 紫色 |
操作步骤
方法一：
1、将放在蔗糖溶液中的组织块取出，放入20g/L蔗糖与OCT包埋剂1：1体积比例混合液中，室温浸泡2h。
2、移入OCT包埋剂中室温浸泡4h，更换新的OCT包埋剂，室温浸泡6h。
3、将标本置于托物台，用恒冷箱切片机切片（一般-22℃），片厚10μm，贴于预处理的载玻片上，-20℃冰箱保存。
方法二：
1、设置恒温冷冻切片机的运行温度在-10℃~-25℃范围内。
2、在样本夹/盘的表面涂一层包埋剂，然后将组织轻轻按压在样本夹/盘上指定的位冒，在组织的周围适当的补加包埋剂，将样本冷冻到适合的切片温度。
注意事项
1、易碎可能是因为冷冻过快或标本太大。
2、组织、包埋块从支架脱落，可能因为包埋剂不足或组织在包埋剂中的位置过偏。
3、组织向前推进但不能顺利切片，可能因为刀和组织未固定紧或刀的角度不合适或组织未完全冷冻。
4、切片卷曲可能因为Anti-roll plate不干净或切片太薄或刀钝。
5、切片撕裂可能因为组织过分冷冻或刀口受损或不洁。
6、切片融化可能因为冷室和刀的温度不正确。
7、刀上有雾，可能因为切片机开盖时间过长。
8、切片粘在anti-roll plate，可能因为温度不正确或anti-roll plate不洁。
9、切片歪向一侧，可能因为刀口钝或受损或anti-roll plate受损。
10、切片从玻片脱落，可能因为没用玻片粘附剂或切的是固定组织、脂肪含量高的标本，操作粗糙。
11、切片水平裂痕，可能因为切片时标本太冷。
12、为了您的安全和健康，请穿实验服并戴一次性手套操作。
OCT Frozen Section Embedding Medium is a water-soluble mixture of polyethylene glycol and polyvinyl alcohol. It is now widely used in immunohistochemistry experiments. Its purpose is to support the tissue during frozen sectioning, improving tissue continuity and reducing folds and fragmentation. Since the embedding medium is water-soluble, it dissolves during the floating process, thus it does not contribute to background staining in subsequent staining procedures. This reagent is intended for research use only and is not suitable for clinical diagnosis or other purposes.
| 项目号 | Color
|
F1508750-01 | Colorless
|
F1508750-02 | Red
|
F1508750-03 | Orange
|
F1508750-04 | Yellow
|
F1508750-05 | Green
|
F1508750-06 | Cyan
|
F1508750-07 | Blue
|
F1508750-08 | Purple |
Procedure
Method One:
Remove the tissue block from the sucrose solution and place it into a 1:1 (v/v) mixture of 20 g/L sucrose and OCT embedding medium. Incubate at room temperature for 2 hours.
Transfer the tissue into pure OCT embedding medium and incubate at room temperature for 4 hours. Then, replace with fresh OCT embedding medium and incubate for an additional 6 hours at room temperature.
Place the specimen onto the specimen holder and section using a cryostat (typically at -22°C) to a thickness of 10 µm. Mount the sections onto pre-treated glass slides and store in a -20°C freezer.
Method Two:
Set the operating temperature of the cryostat within the range of -10°C to -25°C.
Apply a layer of embedding medium onto the surface of the specimen holder/disk. Gently press the tissue onto the designated area of the holder/disk. Apply additional embedding medium appropriately around the tissue. Freeze the sample to the optimal temperature for sectioning.
Notes
Fragility may be caused by overly rapid freezing or excessively large specimen size.
Detachment of the tissue or embedding block from the holder may result from insufficient embedding medium or the tissue being positioned too close to the edge of the medium.
If the tissue advances but does not section smoothly, possible causes include loose fixation of the blade or tissue, incorrect blade angle, or incomplete tissue freezing.
Curling of sections may be due to a dirty anti-roll plate, sections being too thin, or a dull blade.
Tearing of sections may result from excessive freezing of the tissue, a damaged or contaminated blade edge.
Melting of sections may occur if the temperatures of the chamber and blade are incorrect.
Frost/fog on the blade may form if the cryostat lid is left open for too long.
Sections sticking to the anti-roll plate may be caused by incorrect temperature or a dirty anti-roll plate.
Sections skewing to one side may result from a dull or damaged blade edge, or a damaged anti-roll plate.
Sections detaching from the glass slide may occur if slide adhesives were not used, or if the sections are from fixed tissue, specimens high in fat content, or due to rough handling.
Horizontal cracks in sections may be caused by the specimen being too cold during sectioning.
For your safety and health, please wear a lab coat and disposable gloves during operation.
ZJ26F1040093
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