Sample prep for tissues
Tissues must be processed into a monodispersion of cells, typically by mechanical maceration or douncing. Enzymatic digestion (e.g. collagenase, dispase) can be used for connective tissue and Trypsin may be used for macro-dissected tissues (monitor dissolution to single cells). Harvesting viable, monodispersed cells from tissues can be challenging; in some cases nuclei may be preferable. See literature, including the following papers, for additional methods:
- Carpenter et al. Cell-type specific profiling of histone post-translational modifications in the adult mouse striatum. Nature Communications 13 (2022).
- de Bock et al. HOXA9 cooperates with activated JAK/STAT signaling to drive leukemia development. Cancer Discov 8, 616-631 (2018).
- Janssens et al. Automated in situ chromatin profiling efficiently resolves cell types and gene regulatory programs. Epigenetics Chromatin 11 (2018).
- Larsen et al. Establishment, maintenance, and recall of inflammatory memory. Cell Stem Cell 28, 1758-1774 (2021).
- Liu et al. Direct promoter repression by BCL11A controls the fetal to adult hemoglobin switch. Cell 173, 430-442 (2018).
- Miao et al. Glucose dissociates DDX21 dimers to regulate mRNA splicing and tissue differentiation. Cell 186, 80-97 (2023).
- Uyehara & McKay. Direct and widespread role for the nuclear receptor EcR in mediating the response to ecdysone in Drosophila. Proc Natl Acad Sci USA 116, 9893-9902 (2019).