脑组织中的海马取法.doc

取活组织需要玻璃分针，止血钳和冰袋，组织剪。取的时候先断头，从枕骨大孔往前剪开颅骨，完全暴露脑组织，用玻璃分针分离脑组织和颅骨。分离下来的全脑组织置于冰袋上，用玻璃分针从后往上揎起端脑，即可看见海马，左右各一，之间有组织相连，分离的时候要轻柔，尽量避免破坏组织结构。分离下来的组织根据试验安排做后续处理.step 1:This is a dorsal view of a mouses skull, in order to access the brain you can cut along the coronal suture and sagittal suture then pull off both sides of parietal bone and interparietal bone . 这是大鼠头部背面观，为了接触到脑，需沿冠状缝和矢状缝切开，然后把顶骨和枕间骨取下。step 2: Now the skull is open and you can see the brain clearly, in order to get the hippocampus youd better keep the brain in the cranial cavity so the brain will not move when you continue with the following steps 现在头盖骨被打开，你可以清楚地看到大脑，但为了获得海马你最好把大脑保持在颅腔内，这样在你进行下列操作时大脑就不会移动。step 3: In order to expose the hippocampus you need to remove the cerebral cortex covering it. The first incision is at the end of the hemisphere; the incision should be about 0.7mm deep for most adult mouse that you might not hurt the hippocampus while to expose it. The 2nd incision is about 1.5-2mm in front of the first one, this incision you need cut into the lateral ventricle, both of the incisions go to the ventralof the brain and meet there. Now this piece of cortex is free, pull it up, you will see the hippocampus just like in this picture, also you can see the CSF in the opened ventricle 为了看到海马你需要移动覆于其上的大脑皮层。第一个切口在半球的末端，对大多数成年大鼠来说，此切口应约0.7mm深，这样才能保证不伤害到海马。第二个切口在第一个切口前方约1.5-2mm处，此切口需要切入到侧脑室，两个切口在大脑腹侧衔接。现在这块大脑皮层就是游离的了，把它取下来，你就可以看到如图所示的海马，你也可以看到打开的脑室内的CSF（脑脊液）。step 4: Keep working on the other side of the brain pull up both sides of the cortex that covering the hippocampus along the ventricle. Now you can see the dorsal part of the hippocampus. Separate the rest of the hippocampus from the cortex covering it along the surface of the hippocampus towards the ventral part of the hippocampus 继续沿脑室把覆盖在海马上面的两边皮层揭下。现在你能看到海马的背部。把余下的部分沿着海马的表面直到海马的腹部部分从覆与其上的大脑皮层中分离出来.step 5: Keep working on the other side of the brain pull up both sides of the cortex that covering the hippocampus along the ventricle. Now you can see the dorsal part of the hippocampus. Separate the rest of the hippocampus from the cortex covering it along the surface of the hippocampus towards the ventral part of the hippocampus. step 6: Now you need to free the hippocampus from t