Here, we describe an enzymatic dissociation method for mouse brain that prevents dissociation artifacts and lowers technical variations with standardized steps. We tested this protocol on microdissected brain tissue of 3-week- to 24-month-old mice and obtained high-quality scRNA-seq results. For ...
with the 19 clusters obtained from the retained cells (Supplementary Data 2), we were able to match the large majority of the clusters, confirming that the cells removed from each cluster corresponded to stressed cells from each specific cluster, due to high levels of dissociation-related genes...
In terms of the cell dissociation process of tissues for scRNA-seq, certain minor cell types might have faced the risk of being lost. Unsupervised cell collection and a series of deconvoluting algorithm including MNN, MIA and RCTD has been employed to minimize the risk, and to better connect...
To enrich for parenchymal and non-parenchymal liver progenitors, endoderm and associated mesoderm at E7.5 and E8.75 and liver buds at E9.5 and E10.5 were dissected before single-cell dissociation. In addition, the extra-embryonic endoderm- and hepatoblast-specific Afp-GFP mouse line was used to...
Dissociation of fetal tissues Fetal livers and femurs were received in L15 media and processed within 3 h from dissection. Livers were cut in smaller pieces with a scalpel and transferred to a tube containing prewarmed digestion media: RPMI (Gibco) supplemented with 10% FBS (Gibco), penicilli...
China. The clinical information for these samples is provided in Table S6. All samples were processed immediately after being obtained from oral cancer surgery according the standard procedures. According to the manufacturer's instructions, the Human Tumor Dissociation Kit (Miltenyi Biotec; Order no: ...
Single-cell RNA sequencing has been widely adopted to estimate the cellular composition of heterogeneous tissues and obtain transcriptional profiles of individual cells. Multiple approaches for optimal sample dissociation and storage of single cells have
sequencing (snRNA-seq), Lake et al. (2019) have shown the power of this technology to analyse clinical samples bypassing technical limitations in the enzymatic dissociation of the solid tissue and using limiting amounts of sample (Lake et al.2019). Further analysis of receptor-ligand signalling...
We procure tissue from surgical resections and immediately subject it to mechanical and enzymatic dissociation to produce a single-cell suspension. These cells are rapidly loaded into a microfluidic device where they are captured in arrays of microwells (Additional file 1: Figure S1A), subjected to...
2. Tissue dissociation and single-cell sample preparation 3. Drop-seq procedure, single-cell and bulk library generation, and sequencing 4. CITE-seq experiments 5. Immunostainings 6. Processing and analysis of single-cell RNA-seq data ,生信工程师上场,原文写得非常非常详细。 框架: (1) ...