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.
Highlights
The common conception of in vitro diagnostics is intertwined with the idea of a liquid starting sample - blood, saliva, urine, and other starting materials are often the candidates for study, allowing rapid determination of important details about a patient’s health status by investigating metabolomic, proteomic and genomic markers of disease [1]
Cells can be sorted by size directly from bulk solution using a traveling SAWs (TSAWs), governed by a simple equation, adapted from the work of Skowronek et al, which describes whether acoustic radiation drag force from streaming dominates (Eq (2)) [123]
Sample preparation for single-cell analysis is a critical area of importance for future research in order to ensure widespread clinical translation of single-cell sequencing and other single-cell analysis techniques into cancer diagnostics and other workflows
Summary
The common conception of in vitro diagnostics is intertwined with the idea of a liquid starting sample - blood, saliva, urine, and other starting materials are often the candidates for study, allowing rapid determination of important details about a patient’s health status by investigating metabolomic, proteomic and genomic markers of disease [1]. Sources have estimated that the United States spends $8 billion annually on unnecessary repeat biopsy procedures just for breast cancer [2]. This is often because bulk sequencing and histopathological analyses limit the full extent of investigation. The most popular emerging SCA approach for cancer diagnostics is arguably Single-Cell Sequencing (SCS) of nucleic acids (DNA, RNA), which encompasses techniques such as single-cellRNA sequencing (scRNAseq). This is a Generation Sequencing (NGS) approach to characterize the genomes or transcriptomes of individual cells.
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