Cell strainer cap

Manufactured by Thermo Fisher Scientific

Sourced in United States

The Cell Strainer Cap is a laboratory equipment designed to filter and separate cells from a cell suspension. It is a component that can be attached to various conical tubes to facilitate the efficient filtration of cells during experimental procedures.

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Lab products found in correlation

Facsmax cell dissociation solution, by Genlantis (2 mentions) Falcon tube, by Thermo Fisher Scientific (2 mentions) Diva 8, by BD (1 mentions) Lsrii flow cytometer, by BD (1 mentions) Cytoflex flow cytometer, by Beckman Coulter (1 mentions) 96 u bottom plates, by Thermo Fisher Scientific (1 mentions) Anti cd16 32 mab, by BD (1 mentions) Collagenase 4, by Merck Group (1 mentions) Foxp3 transcription factor staining buffer set, by Thermo Fisher Scientific (1 mentions) Dnase 1, by Merck Group (1 mentions) Lsrfortessa, by BD (1 mentions)

6 protocols using cell strainer cap

Dissociation and FACS Sorting of Zebrafish Embryos

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Embryos at 20 hpf were briefly washed in calcium-free Ringer’s solution and deyolked by up and down pipetting. Deyolked embryos were pelleted by 500 ×g centrifugation for 5 min. They were briefly washed with FACSmax cell dissociation solution (Genlantis) and transferred in a 60 mm petri dish with fresh FACSmax solution, then incubated at 28.5°C. Single-cell dissociation was carefully monitored every 5 min and was generally achieved within 30 min of incubation. Efficient dissociation was helped by firmly tapping the petri dish and by gentle pipetting. Single cells were exhaustively washed twice in cold PBS, pelleted and resuspended in 500 μL of cold PBS. Single cells were filtered in a Falcon tube with a cell strainer cap (Fisher Scientific) and placed on ice until cell sorting. Sorting was performed using a BDARIA IIIu FACS with DIVA 8 sofware (BD Biosciences San Jose, CA, USA). GFP expressing cells were identified using a 488 nm laser and a 530/30 BP filter. Cells were collected in cold PBS on ice.

Samarut E., Bekri A, & Drapeau P. (2016). Transcriptomic Analysis of Purified Embryonic Neural Stem Cells from Zebrafish Embryos Reveals Signaling Pathways Involved in Glycine-Dependent Neurogenesis. Frontiers in Molecular Neuroscience, 9, 22.

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Tissue Fixation and Dissection Protocol

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Tissue fixation followed modification of previously published methods (Matthews et al., 2019 (link)) as follows. Heads were carefully removed from the body by pinching at the neck with sharp forceps. Heads were placed in a 1.5 mL tube for fixation with 4% paraformaldehyde, 0.1 M Millonig’s Phosphate Buffer (pH 7.4), 0.25% Triton X-100, and nutated for 3 hour at 4°C. Samples were dissected and samples of the same tissue were grouped into a cell strainer cap (Fisher Scientific #08-771-23) that was cut to fit into 1 well of a 24-well plate containing PBS with 0.25% Triton X-100 (PBT). All subsequent steps were performed on a low-speed orbital shaker at room temperature. Samples were washed at least 5 times 20 min and transferred to PBT. All dissections were performed using this protocol unless otherwise noted.

Jové V., Gong Z., Hol F.J., Zhao Z., Sorrells T.R., Carroll T.S., Prakash M., McBride C.S, & Vosshall L.B. (2020). Sensory Discrimination of Blood and Floral Nectar by Aedes aegypti Mosquitoes. Neuron, 108(6), 1163-1180.e12.

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Single-cell Isolation and Flow Cytometry

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A thousand 123count ebeads (Biosciences #01-1234-42) were added to embryos at 24 hours post-fertilization (hpf). They were briefly washed in calcium-free Ringer’s solution and de-yolked by up and down pipetting. De-yolked embryos were pelleted by 500 × g centrifugation for 5 min. They were briefly washed with FACSmax cell dissociation solution (Genlantis) and transferred in a 60 mm petri dish with FACSmax solution, then incubated at 28.5 °C. Single-cell dissociation was carefully monitored every 5 min and was generally achieved within 30 min of incubation. Efficient dissociation was helped by firmly tapping the petri dish and by gentle pipetting. Single cells were exhaustively washed twice in cold PBS, pelleted and resuspended in 500 μL of cold PBS. Single cells were filtered in a Falcon tube with a cell strainer cap (Fisher Scientific) and placed on ice until counting. Samples were analyzed using a LSRII flow cytometer with DIVA 8 software (BD Biosciences San Jose, CA, USA). GFP expressing cells were identified using a 488 nm laser and a 530/30 BP filter and 123count ebeads with a 561 laser and a 610/20 filter and a 405 laser with a 525/50 filter. 123count ebeads are completely separated from cells on a FSC/SSC dot plot.

Dzieciolowska S., Larroque A.L., Kranjec E.A., Drapeau P, & Samarut E. (2017). The larvicide pyriproxyfen blamed during the Zika virus outbreak does not cause microcephaly in zebrafish embryos. Scientific Reports, 7, 40067.

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Quantitative Analysis of iPSC-Derived Neurons

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Day 30 iPSC-derived neurons were dissociated from plate after 5min Trypsin LE treatments and collected in N2/B27/Rocki medium. After centrifugation at 800 rpm for 5min, supernatant was removed, and the cell pellet was washed once with N2/B27 medium. Neurons were resuspended with 2ml N2/B27/Rocki medium and let cells go through the 5ml round bottom polystyrene tube with cell strainer cap (Fisher scientific) to make single cell suspension. Cells were counted with hemocytometer and 2500 neurons were added to each well on PO/LA coated 96-well plate (transparent bottom black plate). 48hrs later, neurons were fixed with 4% PFA for 10min and stained with TUJ1, Neurofilament antibodies (NF) and Hoechst. The whole picture of each well was taken by TROPHOS plate runner. Images were further analyzed by MetaMorph software to quantify neurite length, neuron body size, number of processes, and count the number of TUJ1-stained cells for each channel.

Wang L., Liu Y., Stratigopoulos G., Panigrahi S.K., Sui L., LeDuc C.A., Glover H.J., Rosa M.C.D., Burnett L.C., Williams D.J., Shang L., Goland R., Tsang S.H., Wardlaw S.L., Egli D., Zheng D., Doege C.A., & Leibel R.L. (2020). Regulation of intracellular signaling and neuron function by Bardet-Biedl Syndrome proteins in patient-specific iPSC-derived neurons.

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Lipid Droplet Quantification Assay

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ZMEL Dark (no fluorescence), ZMEL-GFP, and ZMEL-LD cells were plated on fibronectin-coated six-well plates at a density of 500,000 cells in 1 ml of media per well. At 24 hr after plating, cells were given either 150 µM of BSA or oleic acid with 1 µl of DMSO. At 48 and 72 hr after plating, lipid droplet low and high controls were switched to fresh media with 150 µM of BSA or oleic acid with 1 µl of DMSO. Cells pulsed with oleic acid received fresh media with 150 µM of BSA with either 40 µM Atglistatin, 0.5 µM Auranofin, or 0.5 µM JS-K. At 96 hr after plating, cells were trypsinized, washed with DPBS, and resuspended in DMEM supplemented with 2% FBS, 1x penicillin/streptomycin/glutamine, and 1x GlutaMAX. Cells were stained for viability with 1:1000 DAPI and strained through the Falcon FACS Tube with Cell Strainer Cap (Thermo Fisher, Waltham, USA; catalog #08-771-23). For Lipidtox comparison, cells were given either BSA or indicated concentrations of oleic acid for 24 hr. Cells were trypsinized, washed with DPBS, stained with 1:250 Lipid Deep Red and 1:1000 DAPI for 10 min, and strained through the Falcon FACS Tube with Cell Strainer Cap. Data was acquired via the Beckman Coulter CytoFLEX Flow Cytometer (Beckman Coulter, Miami, USA) and analyzed using FlowJo software (BD Biosciences, San Jose, USA).

Lumaquin D., Johns E., Montal E., Weiss J.M., Ola D., Abuhashem A, & White R.M. (2021). An in vivo reporter for tracking lipid droplet dynamics in transparent zebrafish. eLife, 10, e64744.

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Tumor Dissociation and Flow Cytometry

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Tumors were excised, cut with scissors, and were subsequently incubated with a solution composed of 1 mg/mL collagenase IV (Sigma) and 0.02 mg/mL DNAse I (Sigma) in serum-free RPMI medium for 45 min at 37°C on a heating rotating plate (190 rpm). Cell suspensions were filtered using a 40 μm cell strainer and washed with 20 mL cold FACS buffer (PBS 2% FBS, 5mM EDTA). Cells were centrifuged at 300g for 5 min, and resuspend in cold FACS Buffer, filter through 5 mL round-bottom polystyrene test tubes with cell strainer cap (ThermoFisher) and kept on ice until transfer into 96 U-bottom plates (ThermoFisher) for staining. Following Fc block (anti-CD16/32 mAb (BD Bioscience) for 10 minutes at 4°C), antibodies against cell surface antigens were added (30 minutes incubation at 4°C). For staining of intracellular antigens cells were fixed, permeabilized (eBioscience FoxP3/Transcription Factor Staining Buffer Set) and stained with antibodies following manufacturer’s provided protocol (Supplementary Table S1, “FACS antibodies”). FMO controls were always included as negative controls. To detect OVA-specific CD8⁺ T cells, we used OVA257–264 (SIINFEKL) peptide bound to H-2Kb monoclonal antibody (eBio25-D1.16 (25-D1.16))-APC (from eBioscience, cat #17–5743-82). Flow cytometry was performed on BD LSR Fortessa (BD Bioscience), and FlowJo software was used for analysis.

Jacoberger-Foissac C., Cousineau I., Bareche Y., Allard D., Chrobak P., Allard B., Pommey S., Messaoudi N., McNicoll Y., Soucy G., Koseoglu S., Masia R., Lake A.C., Seo H., Eeles C.B., Rohatgi N., Robson S.C., Turcotte S., Haibe-Kains B, & Stagg J. (2023). CD73 inhibits cGAS-STING and cooperates with CD39 to promote pancreatic cancer. Cancer immunology research, 11(1), 56-71.

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