Preparation of Tissues and Heterogeneous Cellular Samples for Single-Cell Analysis

Abstract

While sample preparation techniques for the chemical and biochemical analysis of tissues are fairly well advanced, the preparation of complex, heterogenous samples for single-cell analysis can be difficult and challenging. Nevertheless, there is growing interest in preparing complex cellular samples, particularly tissues, for analysis via single-cell resolution techniques such as single-cell sequencing or flow cytometry. Recent microfluidic tissue dissociation approaches have helped to expedite the preparation of single cells from tissues through the use of optimized, controlled mechanical forces. Cell sorting and selective cellular recovery from heterogenous samples have also gained traction in biosensors, microfluidic systems, and other diagnostic devices. Together, these recent developments in tissue disaggregation and targeted cellular retrieval have contributed to the development of increasingly streamlined sample preparation workflows for single-cell analysis technologies, which minimize equipment requirements, enable lower processing times and costs, and pave the way for high-throughput, automated technologies. In this chapter, we survey recent developments and emerging trends in this field.

Single-Cell Transcriptomics for Unlocking Personalized Cancer Immunotherapy: Toward Targeting the Origin of Tumor Development Immunogenicity

“…Mechanical methods offer distinct advantages in facilitating prompt and efficient tissue dissociation for scRNA-seq analysis [73]. Moreover, this can overcome the limitations of using fixed or preserved cells, which do not qualify for scRNA-seq analysis due to RNA degradation [74]. On the other hand, the optimization of scRNA-seq protocols can significantly enhance the efficiency and cost-effectiveness of this technology, making it a more reliable and trustworthy single-cell transcriptomics method.…”