Crypt cells were incubated with 10 µM Zinpyr-1 (Santa Cruz Biotechnology) for 10 min at 37°C to stain secretory granules in Paneth cells and filtered with Cell Strainer Snap Cap with 35-µm nylon mesh (BD Falcon). Zinpyr-1+SSClow cells were sorted as Paneth cell–rich fraction using a cell sorter (JSAN; Bay Bioscience), and then Paneth cells identified as Zynpyr-1+ granular cells using a confocal microscope (A1; Nikon) were aspirated one by one using a 50-µm glass micropipette (1-GT50S-5; Nepa Gene) with micromanipulators (MN-4 and MMO-202ND; Narishige) and an electronic pipette (PicoPipet; Nepa Gene).
Cell strainer snap cap
Manufactured by BD
Sourced in United States
The BD Cell Strainer Snap Cap is a laboratory equipment designed to filter and separate cells. It features a durable, reusable mesh filter that can be easily snapped onto various cell culture containers for efficient cell isolation and processing.
Automatically generated - may contain errors
Lab products found in correlation
Zinpyr 1, by Santa Cruz Biotechnology (1 mentions)
Picopipet, by Nepa Gene (1 mentions)
Mmo 202nd, by Narishige (1 mentions)
Propidium iodide solution, by Merck Group (1 mentions)
Round bottom polystyrene test tube, by BD (1 mentions)
Facsaria flow sorter, by BD (1 mentions)
Propidium iodide flow cytometry kit for cell cycle analysis, by Abcam (1 mentions)
Hepes buffer, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Facsaria fusion cell sorter, by BD (1 mentions)
M digestion buffer, by Zymo Research (1 mentions)
Kimble dounce tissue grinder, by Merck Group (1 mentions)
Nuclei ez lysis buffer, by Merck Group (1 mentions)
Facsdiva software, by BD (1 mentions)
8 protocols using cell strainer snap cap
1
Isolation of Paneth Cells from Intestine
2
Cell Cycle Analysis of LN-340 Cells
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LN-340 cells were seeded (0.25 M/10 cm petri dish per condition). Following 5 days pretreatment with JQ1 or DMSO, cells were treated with TMZ and O6BG. After 48 h cells were washed with PBS and fixed with ice-cold 70% ethanol overnight at 4 °C. Subsequently, cell pellets were washed with ice-cold PBS and treated with 1 ml Propidium Iodide solution (20 µg/ml final concentration) (Sigma-Aldrich, P4864-10ml) and RNAse A. Following at least 4 h incubation at 4 °C (protected from light), cells were filtered through 5 ml Round Bottom Polystyrene Test Tubes, with Cell Strainer Snap Cap (Falcon®). Stained and filtered cells were immediately processed with the Gallios II Beckman Coulter (Flow Cytometry Facility—University of Lausanne). Cell cycle distribution was analyzed with the FlowJo software.
3
Nuclei Isolation and FACS Sorting
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Transfer the solution to Falcon™ Round-Bottom Polystyrene Test Tubes through Cell Strainer Snap Cap to filter the nuclei (35-μm cell strainer)
Perform the fluorescence-activated sorting of fixed nuclei ( Representative gating strategy Nuclei were immunostained with an antibody specific to NeuN, a nuclear membrane protein, filtered through a 35-μm cell strainer and sorted on a BD Biosciences FacsAria flow sorter. The right panels illustrate NeuN negative and positive populations. Positive NeuN-PE populations were used for downstream applications.
Collect the NeuN - positive population of nuclei (
After FACS, the solution with nuclei was centrifuged for 10 min at 3200 g and 4°C.
The pellet of fixed nuclei was resuspended in 50 ul of ultra-pure water. The pellet contained 1,3–1,5 m of neuron nuclei. The cell pellet was kept on ice.
4
Cell Cycle Analysis of HepG2 Cells
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HepG2 cells were seeded in six-well plates at a density of 4 × 105 cells/well. After incubation for 24 h, TiO2 and TiO2-PEG NPs were added to all wells except the control wells and incubated for 24 h. The cells were collected by trypsin treatment and then passed through a nylon mesh (Cell Strainer Snap Cap, Falcon, NY, USA) to remove cell clumps. Subsequently, the cells were washed twice with PBS and fixed with 66% ethanol at 4 °C for 2 h, after which they were washed twice with PBS and stained with 200 μl of 1 × propidium iodide (PI) and RNase staining solution (propidium iodide flow cytometry kit for cell cycle analysis, Abcam, Japan) and then incubated at 37 °C for 20 min. Finally, DNA content was assessed using a SP6800 spectral analyzer with 488 nm laser illumination to determine the cell phase.
5
Single Cell FACS Isolation
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For single cell suspension, cells were washed with PBS and incubated with Accutase for 5 min at 37°C, three volumes of 10% FBS (Life Technologies) in PBS was added, and then centrifuged for 3 min at 200 g . Cells were resuspended in FACS buffer [10% FBS, 2 µM EDTA, 0.625 mM HEPES buffer (Sigma-Aldrich) and 10 µM ROCK inhibitor in HBSS (Life Technologies)] at ∼1.5-2 million cells/ml, and transferred to 5 ml Falcon tube with a cell strainer snap cap (Falcon). FACS sorting was performed using and SH800 cell sorter (Sony Biotech), and TdTomato-expressing cells were collected into Eppendorf tubes. At least 2000 cells were collected per sample for RNA isolation.
6
DAPI-based Cell Sorting for Single-cell Analysis
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5 × 106 cell pellets of the NIH3T3 and the B16-F0 were resuspended in 50 μl of 0.1 μg/1 ml of 4,6-diamidino-2-phenylindole (DAPI) (Sigma-Aldrich, D9542-5MG) in 1 ml of phosphate-buffered saline (PBS) (Life Technology, 10010023). Cells were filtered by a Falcon Cell Strainer Snap Cap (Falcon, 352235). DAPI-negative cells (1, 2, 5, 10 and 100 cells) from NIH3T3 and B16-F0 were sorted and collected into 96-well plates pre-loaded with 10 μl of 1× M-Digestion Buffer (Zymo Research, D5020-9) using a BD FACSAria™ Fusion cell sorter (BD Biosciences) with a 100 μm nozzle.
7
Nuclear Isolation and FACS Profiling
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Neuronal nuclei were isolated from dissected brain as described86 (link). Briefly, tissue from the MC and HPC were dissected and homogenized in 2 mL of Nuclei EZ Lysis Buffer (Sigma-Aldrich) using a KIMBLE Dounce Tissue Grinder (Sigma-Aldrich). After the addition of an additional 2 mL of Nuclei EZ Lysis Buffer, samples were incubated at RT for 5 min. Homogenized tissues were then centrifuged at 500g for 5 min. After removing the supernatant, nuclei were resuspended in 4 mL of Nuclei Suspension Buffer (PBS with 100 μg/mL of BSA) and centrifuged at 500g for an additional 5 min. The resulting pellet was then resuspended in 1 mL of Nuclei Suspension Buffer for FACS. Nuclei were strained through Round-Bottom Polystyrene Test Tubes with a 35 μm Cell Strainer Snap Cap (Falcon) and subjected to FACS using a BD FACSAria II Cell Sorter (Roy J. Carver Biotechnology Center Flow Cytometry Facility, University of Illinois Urbana-Champaign, Urbana, IL). Nuclei were collected in 350 μL of RNeasy Plus Kit Lysis Buffer (Qiagen) and at least 15,000 nuclei were collected for each sample.
8
Multiparametric Profiling of hPSCs and BVOs
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Cells were dissociated in enzyme-free dissociation buffer and filtered in a 5 mL round-bottom polystyrene test tube with a cell strainer snap cap (Falcon). hPSCs were stained with FITC-conjugated anti-OCT-4, anti-NANOG, anti-SSEA-4, and anti-TRA-1–60 antibodies, and ECs were stained with FITC-conjugated anti-CD31 and anti-CD144 antibodies. BVOs were detected using APC-conjugated anti-α-SMA and PDGFR-β antibodies. To detect arteries and veins, anti-SOX17 and anti-NR2F2 antibodies, respectively, were used. Staining using antibodies against surface and nuclear factors was performed using DPBS and 1 × perm/wash, respectively. Cell populations were analyzed using an FACSAria instrument with FACS Diva software (BD Biosciences).
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