Project description:Neurons were dissociated and purified from adult (~3 months) mouse brains. We used four strains with genetic markers that enabled live sorting. Thy1-GFP labels a subset of excitatory neurons; VIP-tdTomato, SST-tdTomato and PVALB-tdTomato labels different subsets of inhibitory neurons. The "SAMPLE_ID" sample characteristic is a sample identifier internal to Genentech. The ID of this project in Genentech's ExpressionPlot database is PRJ0016304
Project description:In order to identify the miRNAs in adult Dicer1-CKO Müller glia of the neural retina, we isolated the Müller glia from Rlbp-CreER: Stopf/f-tdTomato/Dicerf/f mice by means of fluorescent activated cell sorting and analyzed their miRNAs using NanoStrings Technologies®. miRNA expression of Dicer1-CKO Müller glia was compared to wild type Müller glia (a re-analyzed sample from GSE94759). We found that all highly expressed miRNAs declined in the Dicer-CKO leading to a disruption in retinal architecture over time.
Project description:Human multipotent mesenchymal stem cells (MSCs), isolated based on their adherence to plastic, show poor growth and differentiation and frequently contain contaminating cells, which limits to clarify their own characteristics. In this study, the identification of two cell surface markers, LNGFR and Thy-1, allowed the prospective isolation of highly purified clonogenic human MSCs. Furthermore, single cell sorting assays followed by in vitro expansion revealed three distinct MSC subpopulations: rapidly-, moderately- and slowly proliferating MSC clones (RPCs, MPCs and SPCs, respectively). While RPCs exhibited robust multilineage differentiation and self-renewal potency, MPCs and SPCs contained a majority of senescent cells and exhibited frequent genome errors. Single cell sorting assays followed by in vitro expansion revealed three distinct MSC subpopulations: rapidly-, moderately- and slowly proliferating MSC clones (RPCs, MPCs and SPCs, respectively). Sample: RPC, MPC and SPC (n=3).
Project description:We have conducted a study to investigate how blocking HLA/LILR affects the activation of immune cells in the tumor microenvironment. We perfused a freshly isolated primary colon cancer biopsy sample with hIgG1 LALAPG or DX17 LALAPG for 48 hours. We then isolated CD45+ leukocytes from the tumor sample using magnetic cell sorting. Subsequently, we processed the resulting tumor-infiltrating lymphocytes (TILs) for single-cell RNA-seq and Cite-seq analysis. We compared the transcriptomes of NK cells, monocytes, and memory T cells under the two different culture conditions
Project description:While mass spectrometry-based single cell proteomics (SCP) is gaining significant momentum, it is largely limited to a few laboratories worldwide. The ability to send samples to specialized core facilities, collaborators, would greatly benefit non-specialists laboratories or those unable to afford the costly instrumentation necessary to perform SCP analysis, and would help to popularize the use of the technology for biological applications. However, no methods have been tested in SCP which allow to “freeze” the proteome state while maintaining cell integrity to enable transfer of single cells suspensions between laboratories and/or prolonged sorting periods using fluorescence-activated cell sorting (FACS). Here we evaluate whether the widespread formaldehyde fixation could be used to maintain cell states enabling shipping between laboratories and on-site sorting, sample processing and mass spectrometry analysis. We demonstrate that short-term fixation using formaldehyde (FA) does not majorly affect protein recovery even in the absence of heating and strong detergent in single-cell proteomics and One-Tip analyses and allow maintaining analytical depth in comparison to classical workflows without fixation. Additionally, we show that fixation preserves drug-induced specific perturbations of protein abundance during cell sorting and sample preparation for SCP analysis. Our study has implication in single-cell and sensitive proteomics and would help provide biologists and other non-specialist researcher access to the technology, while also enables intracellular labelling using antibodies for FACS. This also helps the field expand toward multidimensional analysis where fixing the proteome state at a certain time such as for certain dynamic PTMs is crucial.
2025-02-05 | PXD054445 | Pride
Project description:Dust from waste-sorting plants
| PRJNA925793 | ENA
Project description:cell sorting of Synechococcus population
| PRJNA699323 | ENA
Project description:Spatial Sorting of mouse hepatocytes