Neuronal cell suspensions were passed through a 5μm syringe filter (Millipore), while intestinal cell suspensions (in PBS/2% FBS) were passed through a 35 μm syringe filter and a 40 μm nylon mesh filter (Falcon). Filtered cells were then diluted in the same media as above and sorted using a Sony iCyt Synergy Dual Channel high speed cell sorter (488 nm excitation). Gates were used to eliminate cells with tdTomato and autofluorescence. GFP positive fluorescent events were collected in 1.5 mL Eppendorf tubes containing 10-20 μL PBS/2% FBS, and cells were kept on ice for RNA extraction. Collected positive fluorescent events varied between 20,000 and 60,000 events for intestinal samples, and between 100,000 and 300,000 events for neuronal samples.
40 μm nylon mesh filter
Manufactured by BD
The 40 μm nylon mesh filter is a laboratory equipment designed to filter particles or substances based on size. It has a pore size of 40 micrometers, allowing it to capture and retain materials larger than this size while allowing smaller particles to pass through.
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Lab products found in correlation
2 protocols using 40 μm nylon mesh filter
1
High-Throughput Cell Sorting Protocol
2
Isolation and Quantification of iPSC-Derived Cells
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Non-epithelial cells and epithelial cells were isolated from differentiated iPSCs through manual pipetting. The cells were dissociated using Accutase (Life Technologies) or Accumax and filtered through a 40-μm nylon mesh filter (BD Falcon). The cells (50,000 cells/well) were seeded onto LNE8-coated 96-well plates (Coastar). After incubation for 10–20 min at 37°C, non-adherent cells were washed out with DMEM/F-12 twice. CE maintenance medium was then supplemented. The adherent cells were fixed with 4% paraformaldehyde, washed thrice with TBS, and stained with Hoechst 33342. Stained nuclei were imaged and quantified using a BZ-X810 microscope (Keyence, Osaka, Japan).
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