The Inaugural RaftNotes are Here!
Genome editing, particularly via CRISPR/Cas9 is a powerful method for investigating the role of a given gene in cellular physiology. The method relies on: 1) Reliable introduction (i.e. transfection) of genetic material into cells; 2) Identifying transfection-positive cells; 3) Selecting clones capable of growth and 4) Propagating colonies for analysis of genome edits and downstream functional assays. Here, we present optimized methods using the Cell Microsystems CytoSort Array, a microwell array with releasable single cell isolation features called CellRafts. Using the CytoSort Array, combined with the manual CellRaft System for use with inverted microscopes, we have developed a protocol which represents a more rapid and flexible approach to post-transfection cloning compared to limiting dilution or fluorescence-assisted cell sorting (FACS).
Sorting and isolating single cells is a key sample preparation step in many contemporary workflows including genome editing by CRISPR/Cas9, single cell genomics and differentiation of induced pluripotent stem cells (IPSCs) along with a broad range of other methods. Cell Microsystems has developed the CellRaft AIR™ System to allow imaging-activated cell sorting and isolation of single living cells prior to these downstream analyses. Here, we present optimized methods for staining live cells on the Cell Microsystems CytoSort Array, a microwell array with releasable single cell isolation features called CellRafts. We also describe the use of the automated, bench-top AIR™ System for imaging and isolating vital dye-stained single cells using the CellRaft Technology.