Pietersen Charmaine Y, Lim Maribel P, Macey Laurel, Woo Tsung-Ung W, Sonntag Kai C
Laboratory of Cellular Neuropathology, Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, MA, USA.
Methods Mol Biol. 2011;755:327-43. doi: 10.1007/978-1-61779-163-5_28.
The human brain is an exceptionally heterogeneous structure. In order to gain insight into the neurobiological basis of neural circuit disturbances in various neurologic or psychiatric diseases, it is often important to define the molecular cascades that are associated with these disturbances in a neuronal type-specific manner. This can be achieved by the use of laser microdissection, in combination with molecular techniques such as gene expression profiling. To identify neurons in human postmortem brain tissue, one can use the inherent properties of the neuron, such as pigmentation and morphology or its structural composition through immunohistochemistry (IHC). Here, we describe the isolation of homogeneous neuronal cells and high-quality RNA from human postmortem brain material using a combination of rapid IHC, Nissl staining, or simple morphology with Laser-Capture Microdissection (LCM) or Laser Microdissection (LMD).
人类大脑是一个极其异质性的结构。为了深入了解各种神经或精神疾病中神经回路紊乱的神经生物学基础,以神经元类型特异性的方式定义与这些紊乱相关的分子级联反应通常很重要。这可以通过使用激光显微切割技术,并结合基因表达谱分析等分子技术来实现。为了在人类死后脑组织中识别神经元,可以利用神经元的固有特性,如色素沉着、形态或通过免疫组织化学(IHC)检测其结构组成。在这里,我们描述了结合快速免疫组织化学、尼氏染色,或利用激光捕获显微切割(LCM)或激光显微切割(LMD)的简单形态学方法,从人类死后脑材料中分离出同质神经元细胞和高质量RNA的方法。