Separating Cells:
Before working on single cells, one msut collect them. Some techniques seaprate cells with high pressure and others use tags to enable the identificiton of extraciton of specific cells.
Magnetic fields:
LeviCell uses magnetic fiels to enrich cells without direclty labeling the cells or using any other direct maniipulation of the cells. A cell sample and a paramagnetic solution are introduced to a LeviCell cartridge, directed down a separation channel and subjected to a magnetic density field. In teh separation channel, cells of different types levitate at different heights.
Sequencing single cells:
After collecting cells, one may want to sequence them one by one. This can focus on single cell DNA sequencing or RNA sequencing. This is not an easy task and working with a contract research organization (CRO) is recommened.
Single Cell Discovries (Netherlands) oeprates a laboratory developed specifically for single cell sequencing.
Illumina has a technology where cells are added to barcoded hydrogl templates. Next, the cells are lysed, mRNA is captured on a polyacrylamide poly(T) coated beads and revese transcribed. Full lenght cDNA is sinthesized, amplified, and prepared for sequencing. Library prepration is accomplished using standard illumina paired end cosntructs. It takes 15 hours to go from cells to library. Once processed, samples are analyzed in Illumina’s DRAGEN single cell pipeline and imported to illumina connected multiomics, powered by Partek for intuitive point and click data exploration.
Imaging with omics:
It is one thing to analyze a single cell’s omics and then to also simultaneously determine that cell’s location.
Singular Genomics (San Diego) has a spatial sequencing platform to enable spatially resolved sequencing inside of cells and tissue.