Single-cell sequencing is transforming the field of biological research. As the name implies, this method focuses on the gene expression of individual cells. This approach differs from traditional bulk sequencing methods, which can only give an average view of cell profiles in a tissue sample.
With single-cell sequencing, scientists can identify underlying variability between similar cells that was previously masked, gain insights into rare conditions, and understand subtle cellular differences. These insights are beneficial in fields such as cancer research, developmental biology, immunology, and stem cell research, to name a few.
Still, the promising technology poses some cost and time challenges that require a sophisticated solution like the LUNA-FX7™ Automated Cell Counter.
The Challenges in Single-Cell Sequencing
The complexity of single-cell sequencing starts at the cell isolation stage. This step begins when the tissue or culture arrives at the lab. The individual cells are lysed (broken open) to release their genetic material (DNA or RNA) once isolated. The genetic material is then copied to make it easier for sequencing due to the fact that a single cell contains a small amount of genetic material.
Cell sample quality before this process is crucial, as any compromise can lead to unreliable data. Accurate quantification of cells is the first step in all single-cell sequencing protocols, and that requires reliable tools. Traditional sequencing methods can fail, struggling with accuracy and consistency, especially with low cell concentrations and diverse sample types.
Addressing the Challenges with Automation
The LUNA-FX7™ Automated Cell Counter addresses these challenges head-on. This advanced device can provide accurate cell counts and verify sample quality, ensuring that the investment in single-cell sequencing yields reliable results. By employing sophisticated imaging techniques and precise quantification algorithms, the LUNA-FX7™ excels in evaluating various cell samples, including those with low concentrations and heterogeneous compositions.
Ensuring Sample Quality for Single-Cell RNA-seq
Cell sample quality assessment is a critical step in single-cell RNA sequencing. For instance, B cells isolated from human peripheral blood, when evaluated with the LUNA-FX7™, show high viability and single-cell separation, ideal for downstream library preparation. On the other hand, adherent AsPC-1 cells, if not properly dissociated, form clusters that compromise sample quality. Accurate image-based counting with the LUNA-FX7™ reveals these clusters, highlighting samples unsuitable for further processing.
Evaluating Intact Nuclei for Sequencing
In certain single-cell sequencing applications, isolated cells need to be processed further to obtain intact nuclei. The LUNA-FX7™ proves its utility here as well. For cell lines such as HL60, HeLa, and HEK, the LUNA-FX7™ distinguishes between intact cells, partially isolated nuclei, and fully separated nuclei. By measuring cell size and using high-resolution brightfield images, the device ensures that only high-quality, fully isolated nuclei proceed to sequencing, saving time and resources.
LUNA-FX7™ Is A Game-Changer in Single-Cell Sequencing
Understanding cellular differences is a game-changer for various scientific studies and human life in general, which is why researchers can’t leave anything to chance when it comes to the tools they use in the process. The LUNA-FX7™ Automated Cell Counter’s ability to provide accurate cell counts and verify sample quality addresses critical challenges researchers face and makes the process easier. This can open up new avenues for research and discovery for the scientific community.
Published by: Khy Talara
