A Gravity-Fed Transcardial Perfusion Method for Histologic Analysis of the Mouse Central Nervous System.
A Gravity-Fed Transcardial Perfusion Method for Histologic Analysis of the Mouse Central Nervous System.
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DOI：
10.3791/63386
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发表时间：
2022-01-21
期刊：
影响因子：
--
通讯作者：
Chen XJ
中科院分区：
文献类型：
--
作者：
Rana A;Massa PT;Chen XJ
在这个模型系统中，对分离的小鼠脑和脊髓标本进行组织学分析是评估病理学的常见做法。为了保持这些脆弱组织的形态，常规的做法是通过麻醉动物的心脏插管(经心脏灌流)给动物注射化学固定剂，如多聚甲醛。传统上，小鼠心脏的经心脏灌流依赖于使用蠕动泵或空气压力来输送这一过程所需的生理盐水和固定液。作为这些方法的一种容易使用的替代方法，这项工作演示了使用重力供给法的灌流液输送方法，这种方法使用了大多数五金店都能买到的材料。为了验证这种新的灌流方法，这项工作演示了在大脑和脊髓中灵敏地检测磷酸化α-突触核蛋白所需的所有后续步骤。这些步骤包括固定的脑和脊髓组织的解剖，组织的快速冷冻/包埋和冰冻切片，以及免疫荧光染色。由于这种方法导致固定剂的全身释放，它也可以用于制备其他非神经组织用于组织学分析。一种简便的用于小鼠中枢神经系统组织学分析的重力喂养灌注法。免疫荧光检测磷酸化α-突触核蛋白在帕金森氏病小鼠模型中被证明。这项工作全面描述了经心脏灌流、解剖、组织冷冻/包埋和冷冻切片步骤。
The histologic analysis of brain and spinal cord specimens isolated from mice is common practice for the assessment of pathology in this model system. To maintain the morphology of these delicate tissues, it is routine to administer a chemical fixative such as paraformaldehyde via cannulation of the heart in anesthetized animals (transcardial perfusion). Transcardial perfusion of the mouse heart has traditionally relied on the use of peristaltic pumps or air pressure to deliver both the saline and fixative solutions necessary for this process. As an easily accessible alternative to these methods, this work demonstrates the use of a gravity-fed method of perfusate delivery that uses materials available at most hardware stores. To validate this new perfusion method, this work demonstrate all of the subsequent steps that are necessary for the sensitive detection of phosphorylated α-synuclein in both the brain and spinal cord. Included in these steps are the dissection of the fixed brain and spinal cord tissues, rapid freezing/embedding and cryosectioning of the tissues, and immunofluorescent staining. As this method results in whole-body delivery of the fixative, it may also be used to prepare other non-neuronal tissues for histologic analysis. A convenient gravity-fed perfusion method for histological analysis of the mouse central nervous system. Immunofluorescent detection of phosphorylated α-synuclein is demonstrated in a mouse model of Parkinson’s disease. This work comprehensively describes the transcardial perfusion, dissection, tissue freezing/embedding, and frozen sectioning steps.
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影响因子：
16.2
作者：
Giasson, BI;Duda, JE;Lee, VMY
通讯作者：
Lee, VMY
影响因子：
168.9
作者：
Kalia, Lorraine V.;Lang, Anthony E.
通讯作者：
Lang, Anthony E.
DOI：
10.3791/3564
发表时间：
2012-07-30
期刊：
Journal of visualized experiments : JoVE
影响因子：
--
作者：
Gage, Gregory J;Kipke, Daryl R;Shain, William
通讯作者：
Shain, William
影响因子：
3.2
作者：
Mattson, DL
通讯作者：
Mattson, DL
影响因子：
2.5
作者：
Tao-Cheng, Jung-Hwa;Gallant, Paul E.;Reese, Thomas S.
通讯作者：
Reese, Thomas S.