Here we describe cold-active protease methods for single-cell dissociation of organs and tissues that better preserve the in vivo gene expression patterns.doi:10.1007/978-1-4939-9021-4_5Andrew S. PotterS. Steven Potter
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...
The parameters for simulation of cell signaling are roughly classified into four categories: protein concentration, dissociation constant, diffusion coefficient or transport rate, and enzymatic reaction rate parameters. Classically, these parameters have been measured by in vitro biochemical analyses, which ...
First, we evaluated TotalSeq-B antibodies, custom and MULTISeq LMO hashing on the brain (adapted Miltenyi Biotech Neural Tissue Dissociation kit), spleen, lung, and skin cells where we applied “pre-sort labeling.” We observed that the OCA for brain cells was higher when using lipid hashing...
(Supplementary Table13) were used for scRNA-seq. The HGP was determined retrospectively using H&E slides covering the tumor-liver interface at the margin of sample collection. For depletion of dead cells, specimens underwent single-cell dissociation with the gentleMACs Octo Dissociator (Miltenyi ...
aWorkflow of tumor collection, single-cell dissociation, cell sorting, and computational analysis for scRNA-seq data and whole exome sequencing (WES) data. Among 16 samples subjected to the 10× Genomics method, the 10× 3′v3 method was applied to two samples from two patients, and the 10...
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) ...
2. Tissue dissociation and single-cell sample preparation 收集颌下腺及原发肿瘤标本,置于(DMEM/F12 1:1)、胶原酶1.67 mg/ml (Invitrogen)、透明质酸酶1.33 mg/ml (Sigma)、祛除酶1.67 mg/ml (Invitrogen)中,并用 GentleMacs Dissociator (Miltenyi Biotec) 仪器将其粉碎、解离。细胞悬浮液过70 μm筛,900 ...
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...
(percentage of expressed cells in MCs in CRC) > 0.25, and P value < 0.05. In analyzing expression differences between activated and resting MCs, we excluded 1,514 genes related to mitochondria, heat-shock proteins, ribosomes, and dissociation to eliminate noise and expression artifacts...