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100 μm strainer

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The 100-μm strainer is a laboratory filtration device designed to separate particles or substances larger than 100 micrometers from a liquid or suspension. It is used to remove coarse impurities or unwanted materials from the sample.

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Lab products found in correlation

Percoll, by GE Healthcare (3 mentions) Red cell lysis, by Thermo Fisher Scientific (2 mentions) Pe labeled antimouse podoplanin, by Thermo Fisher Scientific (2 mentions) Apc labeled antimouse lyve 1, by R&D Systems (2 mentions) Dnase 1, by Merck Group (2 mentions) Collagenase d, by Roche (2 mentions) Ack buffer, by Lonza (2 mentions) Mouse cell depletion, by Miltenyi Biotec (2 mentions) Dnase, by Roche (2 mentions) Dnase 1, by Roche (2 mentions) Matrigel, by Thermo Fisher Scientific (2 mentions) Neutral protease, by Worthington (1 mentions) Collagenase 5, by Worthington (1 mentions) Penicillin streptomycin, by Worthington (1 mentions) Roto therm, by Benchmark Scientific (1 mentions) Ethylene diamine tetraacetic acid (edta), by Thermo Fisher Scientific (1 mentions) Rpmi medium, by Thermo Fisher Scientific (1 mentions) Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions) Collagenase 4, by Merck Group (1 mentions) Fetal bovine serum (fbs), by Merck Group (1 mentions)

10 protocols using 100 μm strainer

1

Quantification of Podoplanin and LYVE-1 Expressing Cells

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Lung digests were filtered through 100-μm strainers (Fisher), subjected to red-cell lysis (eBioscience), and stained with PE-labeled antimouse podoplanin (eBioscience) and APC-labeled antimouse LYVE-1 (R&D), with Aqua (Biolegend) viability marker for dead-cell exclusion. Dual PE/APC+ live cells were analyzed by an LSRII (BD) cytometer. The quantity of dual-positive cells as a percentage of total cells was analyzed/plotted and used in statistical analyses comparing lungs from mutant versus control mice.
Johns S.C., Yin X., Jeltsch M., Bishop J.R., Schuksz M., El Ghazal R., Wilcox-Adelman S.A., Alitalo K, & Fuster M.M. (2016). Functional Importance of a Proteoglycan Coreceptor in Pathologic Lymphangiogenesis. Circulation Research, 119(2), 210-221.
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2

Quantitative Flow Cytometry Analysis of Lung Cells

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Lung digests were filtered through 100-μm strainers (Fisher), subjected to red-cell lysis (eBioscience), and stained with PE-labeled anti-mouse podoplanin (eBioscience) and APC-labeled anti-mouse LYVE-1 (R&D), with Aqua (Biolegend) viability marker for dead-cell exclusion. Dual PE/APC+ live cells were analyzed by a LSRII (BD) cytometer. The quantity of dual-positive cells as a percentage of total cells was analyzed/plotted, and used in statistical analyses comparing lungs from mutant versus control mice.
Johns S.C., Yin X., Jeltsch M., Bishop J.R., Schuksz M., El Ghazal R., Wilcox-Adelman S.A., Alitalo K, & Fuster M.M. (2016). Functional Importance of a Proteoglycan Coreceptor in Pathologic Lymphangiogenesis. Circulation Research, 119(2), 210-221.
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3

Isolation of Lymphoid and Intestinal Cells

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Single-cell suspensions of lymphoid organs were prepared by mechanic disruption through 100-μm strainers (Thermo Fisher Scientific). Red blood cells were lysed using Ammonium-Chloride-Potassium (ACK) buffer (Lonza, Walkersville, MD). Small intestinal and colonic lamina propria cells were prepared as previously described (Yuan et al., 2017 (link)). Briefly, the intestine was physically emptied, opened longitudinally and sliced into 0.2–0.5 cm pieces, followed by incubating in 20 mL PBS containing 5 mM EDTA at 37°C on a shaker (200 x rpm) for 15 min. The intestine fragments were further digested in 20 mL RPMI 1640 containing 2% FBS, Collagenase D (1 mg/ml) (Roche) and DNase I (0.1 mg/ml) (Sigma-Aldrich) at 37°C on a shaker (200 x rpm) for 45 min. Samples were collected by passing through 70 μm cell strainers. The cells were then purified using a 40/80% Percoll (GE Healthcare Life Science) gradient by centrifugation (900 x g, 20 min, no break at room temperature). The interphase was harvested and washed. The cells were resuspended in PBS containing 0.2% bovine serum albumin or medium containing fetal calf serum before extra/intracellular staining.
Yang B.H., Wang K., Wan S., Liang Y., Yuan X., Dong Y., Cho S., Xu W., Jepsen K., Feng G.S., Lu L.F., Xue H.H, & Fu W. (2019). TCF1 and LEF1 Control Treg Competitive Survival and Tfr Development to Prevent Autoimmune Diseases. Cell reports, 27(12), 3629-3645.e6.
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4

Isolation of Lymphoid and Intestinal Cells

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Single-cell suspensions of lymphoid organs were prepared by mechanic disruption through 100-μm strainers (Thermo Fisher Scientific). Red blood cells were lysed using Ammonium-Chloride-Potassium (ACK) buffer (Lonza, Walkersville, MD). Small intestinal and colonic lamina propria cells were prepared as previously described (Yuan et al., 2017 (link)). Briefly, the intestine was physically emptied, opened longitudinally and sliced into 0.2–0.5 cm pieces, followed by incubating in 20 mL PBS containing 5 mM EDTA at 37°C on a shaker (200 x rpm) for 15 min. The intestine fragments were further digested in 20 mL RPMI 1640 containing 2% FBS, Collagenase D (1 mg/ml) (Roche) and DNase I (0.1 mg/ml) (Sigma-Aldrich) at 37°C on a shaker (200 x rpm) for 45 min. Samples were collected by passing through 70 μm cell strainers. The cells were then purified using a 40/80% Percoll (GE Healthcare Life Science) gradient by centrifugation (900 x g, 20 min, no break at room temperature). The interphase was harvested and washed. The cells were resuspended in PBS containing 0.2% bovine serum albumin or medium containing fetal calf serum before extra/intracellular staining.
Yang B.H., Wang K., Wan S., Liang Y., Yuan X., Dong Y., Cho S., Xu W., Jepsen K., Feng G.S., Lu L.F., Xue H.H, & Fu W. (2019). TCF1 and LEF1 Control Treg Competitive Survival and Tfr Development to Prevent Autoimmune Diseases. Cell reports, 27(12), 3629-3645.e6.
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5

Isolation and Culture of PDX Tumors

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Tumor bearing mice were euthanized in accordance with institutional Animal Care and Use Committee approval (#20027-H). PDX were harvested and cut into 2-mm pieces, connective tissue, blood vessels and necrotic tissue was discarded and then transferred into 50-mL Falcon tubes with RPMI, 2% Penicillin/Streptomycin, collagenase 5 (Worthington 1 mg/mL), neutral protease (Worthington 0.5U/ml) and DNAse (Roche, 1 μg/mL), and digested while rotating at 37°C until digestion was complete (Benchmark scientific Roto-therm). All following steps were done on ice or at 4°C. The digested tissue was passed through a 200-μm (Pluriselect) strainer using a syringe plunger for remaining pieces, and then through a 100-μm strainer (Fisher). The cells were spun down at 321×g for 5 minutes at 4°C and the pellet depleted of red blood cells by a 10-second exposure to 1 mL of water followed by the addition of 49 mL of PBS. The cells were counted and plated on collagen coated plates for in vitro experiments. For in vitro experiments, the cells from PDX tumors were subjected to mouse cell depletion according to the manufacturer’s instructions (Miltenyi Biotec).
Neumayer C., Ng D., Jiang C.S., Qureshi A., Lalazar G., Vaughan R, & Simon S.M. (2022). Oncogenic Addiction of Fibrolamellar Hepatocellular Carcinoma to the Fusion Kinase DNAJB1-PRKACA. Clinical Cancer Research, 29(1), 271-278.
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6

Isolation of Intestinal Lymphocytes and Dendritic Cells

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MLN lymphocytes were obtained by mechanical disruption and filtered through a 70μm cell strainer in staining buffer [2% FBS (Sigma), 1mM EDTA (Invitrogen), 500ml PBS (GIBCO)]. DC isolation from the small intestinal draining MLN was performed by digestion with collagenase IV (0.5 mg/mL, Sigma–Aldrich) and DNase I (12.5 μg/mL, Roche) diluted in R10 media (RPMI 1640 + 10% FCS) for 40 min at room temperature, followed by mechanical disruption and filtering through a 70μm cell strainer. For the isolation of small intestinal lamina propria (SILP) lymphocytes, the intestine was opened longitudinally after removal of Peyer’s Patches and fat and cut into small pieces. Tissue pieces were incubated on a shaker in HBSS containing 2% FBS and 2mM EDTA at 37°C for three rounds (first round 10 mins, followed by two rounds of 15 mins each). Buffer exchange was performed by discarding the supernatant through a nylon mesh, retaining the tissue pieces. Subsequently, the remaining tissue was digested in RPMI medium (Gibco) containing 10% FBS, 58μg/ml Liberase TM (Roche) (amounting to 0.3 WünschU/ml) and 30μg/ml DNase I (Roche) on a magnetic stirrer for 20 mins at 37°C. Upon digestion, the cell suspension was filtered through a 100μm strainer (Fisher Scientific) and lymphocytes were enriched by density gradient centrifugation with 40%/70% Percoll (GE Healthcare).
Muleta K.G., Ulmert I., Hamza K.H., van Dijl S., Nakawesi J, & Lahl K. (2022). Rotavirus-Induced Expansion of Antigen-Specific CD8 T Cells Does Not Require Signaling via TLR3, MyD88 or the Type I Interferon Receptor. Frontiers in Immunology, 13, 814491.
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7

Immunofluorescence Assay of hiPSC-derived NSCs

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An additional immunofluorescence assay with hiPSC-derived NSCs was performed on two samples each of Matrigel-coated PEDOT: PSS ink and plastic, as control. The ink was printed in a square shape of 8 × 8 mm, with the combination of parameters: Ønozzle = 300 μm, s = 25 mm/min, A = 40 sccm, S = 80 sccm, T = 40°C, and for 20 layers. Post-printing curing was performed in a thermal oven (Heraeus) at T = 150°C for 8 min. Later, Matrigel (Thermo-Fisher Scientific) was drop casted on each sample for 1 h at 37°C. NSCs were filtered with a 100 μm strainer (FisherScientific) in order to collect a single cell suspension. Cells were then plated at a concentration of 5 × 104 cells cm−2 and later deposited on the substrates. The samples were incubated for 30 min and eventually filled with an adequate volume of Neural Expansion Medium (Thermo-Fisher Scientific). The well-plate was finally incubated at 37°C in an atmosphere at 5% CO2, over a period of 4 days. The same procedure for the NTE substrates was then applied to these specimens.
Seiti M., Ginestra P.S., Ferraro R.M., Giliani S., Vetrano R.M., Ceretti E, & Ferraris E. (2022). Aerosol Jet® Printing of Poly(3,4-Ethylenedioxythiophene): Poly(Styrenesulfonate) onto Micropatterned Substrates for Neural Cells In Vitro Stimulation. International Journal of Bioprinting, 8(1), 504.
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8

Cell Compatibility Evaluation of PAN+CNT Fibres

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Human-iPSC-derived NSCs [65, (link)66] (link) were used to perform cell compatibility tests on three samples of PAN+CNT carbonized fibres. The substrates were positioned in a 24-well plate then washed with PBS and sterilized in an autoclave. A Matrigel (Thermo-Fisher Scientific) coating was applied for 1 h at 37 °C. NSCs were passed through a 100 μm strainer (Fisher Scientific) to obtain a single cell suspension and plated at 5×10 4 cells cm -2 . A concentrated cell suspension was deposited on the samples and incubated for 20 min before filling the 24-well plate with the appropriate volume of StemPro NSC SFM (Thermo-Fisher Scientific). Cells were maintained at 37 °C in an atmosphere containing 5% CO 2 . After 5 days in culture, the cells were fixed and permeabilized using Fix&Perm-Reagent kit (SIC), blocked for 45 min with iBindTM Buffer solution (Invitrogen) and stained with Phalloidin, specific for the cytoskeletal component of the cells. Cell nuclei were counterstained with Hoechst 33342 to show the living cells. Samples were observed with an inverted fluorescence microscope (Olympus IX70).
Seiti M., Ginestra P., Ferraro R.M., Ceretti E, & Ferraris E. (2020). Nebulized jet-based printing of bio-electrical scaffolds for neural tissue engineering: a feasibility study. Biofabrication, 12(2).
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9

Isolation and Characterization of Immune Cells from Tumor Samples

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Resected tumors or the contralateral hemisphere were digested with 1 mg ml–1 collagenase IV (StemCell Technologies) and 1 mg ml–1 DNase I (Roche) for 15 min at 37 °C. Samples were strained through a 100-μm strainer (Fisherbrand) and washed with PBS. Cells were stained with a LIVE/DEAD Fixable Blue Dead Cell Stain kit (Thermo Fisher Scientific) for 10 min on ice, treated with FcR blocking reagent (Miltenyi Biotec) diluted 1:50 for 15 min on ice and stained with 1:100 APC-conjugated anti-CD11b (BioLegend, clone M1/70) for 20 min to exclude phagocytic cells. Samples were fixed overnight with a eBioscience FoxP3 transcription factor fixation kit (Thermo Fisher Scientific) and analyzed with a BD LSRII Fortessa (BD Biosciences) in PBS.
Watson D.C., Bayik D., Storevik S., Moreino S.S., Sprowls S.A., Han J., Augustsson M.T., Lauko A., Sravya P., Røsland G.V., Troike K., Tronstad K.J., Wang S., Sarnow K., Kay K., Lunavat T.R., Silver D.J., Dayal S., Joseph J.V., Mulkearns-Hubert E., Ystaas L.A., Deshpande G., Guyon J., Zhou Y., Magaut C.R., Seder J., Neises L., Williford S.E., Meiser J., Scott A.J., Sajjakulnukit P., Mears J.A., Bjerkvig R., Chakraborty A., Daubon T., Cheng F., Lyssiotis C.A., Wahl D.R., Hjelmeland A.B., Hossain J.A., Miletic H, & Lathia J.D. (2023). GAP43-dependent mitochondria transfer from astrocytes enhances glioblastoma tumorigenicity. Nature Cancer, 4(5), 648-664.
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10

Isolation and Purification of PDX Tumor Cells

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The 2mm pieces of tissue were placed into 50mL Falcon© tubes with RPMI, collagenase 4 (Worthington 1 mg/ml) and DNAse (Roche, 1 μg/ml), and digested while rotating at 37C until digestion was complete (Benchmark scientific Roto-therm). All following steps were done on ice or at 4C. The digested tissue was passed through a 200μm (Pluriselect) strainer using a syringe plunger for remaining pieces, and then through a 100μm strainer (Fisher). The cells were spun down at 300xg for 5 minutes at 4C and the pellet depleted of red blood cells by a 10 sec exposure to 1ml of water followed by the addition of 49mL of PBS. The cells were counted and either implanted into mice or used for in vitro experiments. For in vitro experiments the cells from PDX tumors were subjected to mouse cell depletion according to the manufacturer’s instructions (Miltenyi Biotec).
Lalazar G., Requena D., Ramos-Espiritu L., Ng D., Bhola P.D., de Jong Y.P., Wang R., Narayan N.J., Shebl B., Levin S., Michalidis E., Kabbani M., Vercauteren K.O., Hurley A.M., Farber B.A., Hammond W.J., Saltsman JA I.I.I., Weinberg E.M., Glickman J.F., Lyons B.A., Ellison J., Schadde E., Hertl M., Leiting J.L., Truty M.J., Smoot R.L., Tierney F., Kato T., Wendel H.G., LaQuaglia M.P., Rice C.M., Letai A., Coffino P., Torbenson M.S., Ortiz M.V, & Simon S.M. (2021). Identification of Novel Therapeutic Targets for Fibrolamellar Carcinoma Using Patient Derived Xenografts and Direct from Patient Screening. Cancer discovery, 11(10), 2544-2563.
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