Nichterwitz Susanne, Benitez Julio Aguila, Hoogstraaten Rein, Deng Qiaolin, Hedlund Eva
Department of Neuroscience, Karolinska Institutet, Retzius v. 8, 171 77, Stockholm, Sweden.
Department of Cell and Molecular Biology, Karolinska Institutet, Berzelius v. 35, 171 77, Stockholm, Sweden.
Methods Mol Biol. 2018;1649:95-110. doi: 10.1007/978-1-4939-7213-5_6.
LCM-seq couples laser capture microdissection of cells from frozen tissues with polyA-based RNA sequencing and is applicable to single neurons. The method utilizes off-the-shelf reagents and direct lysis of the cells without RNA purification, making it a simple and relatively cheap method with high reproducibility and sensitivity compared to previous methods. The advantage with LCM-seq is also that tissue sections are kept intact and thus the positional information of each cell is preserved.
激光捕获显微切割测序(LCM-seq)将从冷冻组织中进行细胞的激光捕获显微切割与基于polyA的RNA测序相结合,适用于单个神经元。该方法使用现成的试剂,直接裂解细胞而无需进行RNA纯化,与之前的方法相比,它是一种简单且相对便宜的方法,具有高重现性和高灵敏度。LCM-seq的优势还在于组织切片保持完整,因此每个细胞的位置信息得以保留。