Technical Note, Gene Expression, Last Modified on August 9, 2021, Permalink
Technical Note - Guidelines for Accurate Target Cell Counts Using 10x Genomics® Single Cell Solutions
This Technical Note presents an overview of best practices for cell handling and counting that will increase the accuracy of cell counts and minimize the variability in the number of loaded cells. 10x Genomics Single Cell Protocols require suspensions of viable, single cells as input. Accurate counting and qualification of cells during the workflow of the Single Cell 3’ v2 and Single Cell V(D)J Protocol is essential to determine the number of cells to load on to the microfluidic chip for analysis on the system. While manual (e.g. hemocytometer) and automated (e.g. Countess II Automated Cell Counter) counting methods can be used to report cell concentrations, additional factors should be considered that can affect the final count.
FOR USE WITH
This technical note presents guidelines for counting cells in the range of 700-1200 cells/µl. If using the Single Cell 3' LT v3.1 (low throughput) application, ensure cells are counted as indicated in this protocol and then diluted to the LT specific optimal loading concentration of 100-600 cells/µl using the Cell Dilution Overview in the LT User Guide.