Question: Which tissue dissociation protocols are supported for use with the Flex Gene Expression assay?
Answer: Optimized tissue dissociation protocols for other single-cell assays can be used in the Flex Gene Expression assay. Tissue dissociation protocols that have been previously tested for compatibility with the Single Cell Gene Expression assay (i.e., 3’ GEX) are expected to perform similarly in the Flex Gene Expression assay. If these isolation methods/protocols yielded good results for 3’ GEX, they are expected to perform similarly in Flex Gene Expression.
Fresh Tissue
Fresh tissue can be used in the Flex Gene Expression assay by following the 10x Demonstrated Protocol for Tissue Fixation & Dissociation Demonstrated Protocol. This protocol is preferred if you do not have an optimized tissue dissociation protocol for single cell suspensions. The fixation of whole tissues for use in the Flex Gene Expression assay is not be supported. Tissue samples must first be chopped into smaller pieces before fixation for optimal assay performance. This step is required for uniform fixation and permeabilization.
Alternatively, fresh tissue can be dissociated into single-cell suspensions using the 10x Demonstrated Protocol: Tumor Dissociation for Single Cell RNA Sequencing. To thaw these cryopreserved samples, we recommend following the 10x Demonstrated Protocol: Thawing Dissociated Tumor Cells for Single Cell RNA Sequencing.
Frozen Tissue
Frozen tissue can be used in the Flex Gene Expression assay by following the 10x Demonstrated Protocol for Tissue Fixation & Dissociation Demonstrated Protocol This protocol is the preferred sample preparation method for frozen tissue for optimal assay performance. Alternatively, nuclei can be isolated from frozen tissue using the Chromium Nuclei Isolation Kit.
We recommend performing pilot experiments to optimize sample preparation methods for specific tissue types before committing to large-scale studies.
Products: Flex Gene Expression