Methanol fixation is the method of choice for droplet-based single-cell transcriptomics of neural cells

甲醇固定法是基于液滴的单细胞神经细胞转录组学研究的首选方法。

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作者:Ana Gutiérrez-Franco #,Franz Ake #,Mohamed N Hassan,Natalie Chaves Cayuela,Loris Mularoni,Mireya Plass

Abstract

The main critical step in single-cell transcriptomics is sample preparation. Several methods have been developed to preserve cells after dissociation to uncouple sample handling from library preparation. Yet, the suitability of these methods depends on the cell types to be processed. In this project, we perform a systematic comparison of preservation methods for droplet-based single-cell RNA-seq on neural and glial cells derived from induced pluripotent stem cells. Our results show that while DMSO provides the highest cell quality in terms of RNA molecules and genes detected per cell, it strongly affects the cellular composition and induces the expression of stress and apoptosis genes. In contrast, methanol fixed samples display a cellular composition similar to fresh samples and provide a good cell quality and little expression biases. Taken together, our results show that methanol fixation is the method of choice for performing droplet-based single-cell transcriptomics experiments on neural cell populations.

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