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Single-cell omics refers to the comprehensive analysis of molecular profiles at the resolution of individual cells, offering unprecedented insights into cellular heterogeneity and dynamics. Traditional bulk omics techniques analyse pooled samples of cells and thus mask the variability that exists between individual cells. In contrast, single-cell approaches enable the dissection of variability across various molecular layers, such as transcriptomics, genomics, proteomics, and lipidomics, and provide more granular insights into complex biological processes that govern cellular function and behaviour.
Collectively, single-cell omics is transforming our ability to investigate cellular diversity, development, and disease at an unprecedented resolution.
In single-cell transcriptomics, high-throughput sequencing technologies like RNA-seq are employed to measure gene expression at the single-cell level, enabling the identification of rare cell types, differentiation states, and responses to environmental stimuli.
Single-cell lipidomics examines lipid profiles at the cellular level, offering insights into membrane dynamics, metabolic processes, and signaling pathways. It is an emerging field with challenges including the need for highly sensitive techniques, efficient lipid extraction methods, and accurate data interpretation due to the small amounts of material in each cell.
Single-cell proteomics involves the analysis of proteins, often using mass spectrometry or antibody-based techniques, to study protein abundance, modifications, and interactions in individual cells. This approach can reveal how protein networks are reprogrammed in response to cellular states or diseases.
Single-cell genomics focuses on the analysis of DNA within individual cells, providing insights into genomic variability, mutations, and copy number alterations that may be missed in bulk sequencing. This is particularly valuable in fields like cancer research, where tumor cells exhibit significant genomic heterogeneity.
Successfully establishing single-cell omics workflows entails several critical steps, including the isolation of verified single cells and downstream sample processing. iotaSciences’ scPicking Platform greatly simplifies the single-cell isolation process by automating all tedious liquid handling steps, while reliably assuring monoclonality utilizing miniature cell-culture chambers. In addition, selected single cells are automatically transferred into various types of containers compatible with a range of single-cell omics applications, such as well-plates, PCR plates, tubes and vials for mass spectrometry. Samples can optionally be cooled throughout the workflow to preserve precious material and ensure the integrity of the various solutes.
Creating a single-cell suspension from a heterogeneous cell sample involves mechanically or enzymatically dissociating a bulk cell population into individual cells. This process may include the use of enzymes such as trypsin to break down the extracellular matrix or cell-cell adhesions. Gentle pipetting is used to further separate the cells. After dissociation, the suspension may be filtered through a mesh or strainer to remove clumps, ensuring a uniform population of single cells. The final cell suspension is then used to isolate individual cells.
After establishing a single-cell suspension, the next step is to isolate individual cells. Traditional manual isolation methods are tedious and unreliable for ensuring successful isolation of single cells.
The automated scPicking Platform solves these problems by using miniature culture chambers for easy and reliable monoclonality verification. It automates all pipetting tasks, such as plating single cells as well as transferring selected single cells into vessels compatible with the respective application.
The different types of omics applications utilize different containers for processing. For example, single-cell genomics and transcriptomics may use well-plates or tubes, while mass-spec driven approaches require well-plates or distinct mass spectrometry compatible vials.
The scPicking Platform transfers single cells flexibly into various containers compatible with a variety of downstream applications, including well-plates, PCR plates, tubes as well as vials suitable for mass spectrometry.
Once single cells have been transferred into assay-compatible containers, the samples will need further processing. This may involve snap-freezing cells or direct lyses with selected reagents. Depending on the assay, samples will have to undergo various steps prior to acquiring data and analyses. For example, for single-cell transcriptomics, RNA from lysed single cells will need to be barcoded and reverse-transcribed prior to NGS library preparation and sequencing.
Want to find out more about our single-cell platforms? Contact one of our knowledgeable experts today, visit our resources page, or request a demo to see the platforms in action.