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Addgene: Isolating a Monoclonal Cell Population by ...

Addgene: Isolating a Monoclonal Cell Population by ...

Aug 01, 2016 Isolate cells from the stable cell pool by trypsinization and break up any cell clumps into individual cells by passing several times through a serological pipet or by passing through a 0.40 m cell strainer mesh. Quantitate the cell concentration in this homogenized cell solution with a hemocytometer or other cell counter.

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Protocol for Reducing Cell Clumping in Single Cell ...

Protocol for Reducing Cell Clumping in Single Cell ...

Centrifugation. How to Reduce Cell Clumping in Single-Cell Suspensions Using DNase I. ... Rinse the sample tube three times with culture medium or buffer containing 2% FBS, then pass through the strainer. The single-cell suspension is now ready for cell counting and further downstream applications such as cell isolation.

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Mapping transcriptomic vector fields of single cells: Cell

Mapping transcriptomic vector fields of single cells: Cell

Feb 01, 2022 Dynamo is a computational framework for gaining insights into dynamic biological processes, such as human hematopoiesis, from time-resolved single-cell RNA-seq data. Dynamo constructs transcriptomic vector fields from single-cell data and enables predictive modeling of cell-state regulatory mechanisms, perturbation outcomes, and optimal paths for cell-state …

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Functional visualization of NK cell-mediated killing of ...

Functional visualization of NK cell-mediated killing of ...

Feb 03, 2022 A single-cell suspension of the lung cells was generated by mincing the resected lungs with scissors and incubating the minced tissue in RPMI containing 200 U/ml collagenase type IV and 5 U/ml DNase I for 30 min at 37 C. The lysed tissue was then passed through a ϕ40-μm cell strainer.

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Engineered virus-like particles for efficient in vivo ...

Engineered virus-like particles for efficient in vivo ...

Jan 20, 2022 After centrifugation, the supernatant was removed, and nuclei were resuspended in 1-2 mL of NSB, passed through 35-μm cell strainer, followed by flow sorting using the Sony MA900 Cell Sorter (Sony Biotechnology) at the Broad Institute flow cytometry core. See Figure S5A for example FACS gating. Nuclei were sorted into DNAdvance lysis buffer ...

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Translational control of E2f1 regulates the Drosophila ...

Translational control of E2f1 regulates the Drosophila ...

Jan 24, 2022 E2F transcription factors are master regulators of the eukaryotic cell cycle. In Drosophila, the sole activating E2F, E2F1, is both required for and sufficient to promote G1→S progression.E2F1 activity is regulated both by binding to RB Family repressors and by posttranscriptional control of E2F1 protein levels by the EGFR and TOR signaling pathways.

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Nuclei isolation of multiple brain cell types for omics ...

Nuclei isolation of multiple brain cell types for omics ...

Jan 25, 2021 Pass nuclei (from Step 46 or 65) through a 35- m cell strainer snap cap into a new FACS tube. 67 To counterstain the nuclei with 0.5 g ml −1 DAPI, add 1 l 0.5 mg ml −1 DAPI working stock ...

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A single-cell atlas of the normal and malformed human ...

A single-cell atlas of the normal and malformed human ...

Jan 27, 2022 Cell suspensions were filtered through a sterile 40-μm cell strainer (Fisher Scientific) to isolate undigested debris. Cells contained within the flowthrough were collected by centrifugation at 500g at 5 min and the supernatant carefully aspirated. To lyse any residual erythrocytes, the cell pellet was resuspended in Gibco ACK lysing buffer ...

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Ultra-high-throughput single-cell RNA sequencing and ...

Ultra-high-throughput single-cell RNA sequencing and ...

May 31, 2021 Cell pellets were collected by centrifugation at 300g for 5 min, PBS was completely removed, and cells were resuspended in 150 l of ice-cold methanol. The plate was covered with an aluminum seal ...

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Single-cell analyses reveal key immune cell subsets ...

Single-cell analyses reveal key immune cell subsets ...

Oct 14, 2021 Zhang et al. combine single-cell RNA-seq, TCR-seq, and ATAC-seq to investigate immune cell dynamics in the tumor microenvironment and peripheral blood of patients with TNBC treated with paclitaxel or paclitaxel plus atezolizumab, revealing immune features of responders and nonresponders, the mechanisms and intertwined effects of paclitaxel and atezolizumab in …

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Bone Marrow-Derived Macrophages (BMM): Isolation and ...

Bone Marrow-Derived Macrophages (BMM): Isolation and ...

Pipet the bone marrow cells up and down to bring the cells into single-cell suspension. 10. Pass the cells through a cell strainer. Wash the strainer with another 5 mL of lymphocyte medium. 11. Count the bone marrow cells using a hemacytometer. Adjust the concentration to 2 10 6 cells/mL in BMM medium. 12.

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Guidelines for the use of flow cytometry and cell sorting ...

Guidelines for the use of flow cytometry and cell sorting ...

These guidelines are a consensus work of a considerable number of members of the immunology and flow cytometry community. They provide the theory and key practical aspects of flow cytometry enabling immunologists to avoid the common errors that often ...

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