Thursday, May 19, 2022

A new way to boost the efficiency of single-cell RNA sequencing

DisCo: a powerful tool that can “read” the genetic profile of an individual cell.

Single-cell RNA sequencing or ‘scRNA-seq’ allows scientists to study the expression of genes in an individual cell within a mixed population. This technique involves capturing the RNA of a single cell and sequencing it after multiple molecular conversion reactions.

The single-cell RNA sequencing technique has become the gold standard for defining cell states and phenotypes. The technique has transformed the ability to resolve cellular properties across systems. However, the problem is that they are currently tailored toward large cell inputs.

But, this is not a trivial problem as the method requires over a thousand cells for a useful measurement. This renders them inefficient and costly when processing small, individual samples such as small tissues or patient biopsies.

To efficiently process samples with fewer cells, bioengineers at EPFL have developed a new method called DisCo. DisCo stands for ‘deterministic, mRNA-capture bead and cell co-encapsulation droplets system.’

This newly developed method uses machine vision to detect cells actively and capture them in droplets of oil and beads. This approach allows for continuous operation and renders scaling and serial processing of cell samples highly cost-efficient.

In their study, scientists found that DisCo This approach allows for continuous operation and renders scaling and serial processing of cell samples highly cost-efficient. It also controls sorting through combined machine-vision and multilayer microfluidics.

Scientists tested DisCo on the small chemosensory organs of the Drosophila fruit fly and individual intestinal crypts and organoids. The latter are tiny tissues grown in culture dishes closely resembling actual organs – a field that EPFL has been spearheading for years.

Using DisCo, scientists analyzed individual intestinal organoids at different developmental stages. The approach painted a fascinating picture of heterogeneity in the organoids, detecting various distinct organoid subtypes some had never been identified before.

Professor Bart Deplancke at EPFL’s School of Life Sciences said, “Our work demonstrates the unique ability of DisCo to provide high-resolution snapshots of cellular heterogeneity in small, individual tissues.”

Journal Reference:

  1. Johannes Bues, Marjan Biočanin et al. Deterministic scRNA-seq captures variation in intestinal crypt and organoid composition. DOI: 10.1038/s41592-021-01391-1

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