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3.2K Views
•
07:14 min
October 21st, 2021
DOI :
10.3791/63007-v
Chapters
0:04
Introduction
0:50
Perfusion
2:01
Dissection
3:11
Adult Brain Dissociation
4:15
Debris Removal
5:28
Live and Dead Staining
6:00
Results: Flow Cytometry Analysis Following Automated Mechanical Dissociation and Enzymatic Digestion of Mouse Brain Hippocampal Tissue
6:51
Conclusion
Transcript
成功解离少量神经组织可以使实验室获得对治疗效果,细胞功能以及疾病和治疗作用机制的结构特异性见解。这种神经解离方案始终如一地产生高度可行和实际的单细胞悬浮液。此外,我们可以处理的样品只是商业试剂盒所针对样品的一小部分。
适当的规划和准备是该技术成功的关键。运行几轮练习以熟悉协议将是有益的。麻醉一只六个月大的雌性C57BL / 6J小鼠后,捏住下腹部并使用镊子抬起皮肤。
然后用剪刀,穿过皮毛和皮肤到肋骨底部。做两个对角线切口,从胸腔下方开始,向每个肩膀移动。然后小心地切
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Summary
该神经细胞解离方案适用于起始材料量低的样品,并产生用于下游分析的高活性单细胞悬浮液,并具有可选的固定和染色步骤。
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