70 μm cell strainer

Manufactured by Celltreat

Sourced in United States

The 70 μm cell strainer is a laboratory tool used for filtration and separation purposes. It features a mesh with pore size of 70 micrometers, which allows for the effective removal of larger cell clumps or debris from cell suspensions while retaining the desired cell population.

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Lab products found in correlation

Hanks balanced salt solution (hbss), by Thermo Fisher Scientific (1 mentions) Nexcelom cellometer auto t4 cell counter, by Revvity (1 mentions) Lsrfortessa x 20, by FlowJo (1 mentions) Countbright absolute counting beads, by Thermo Fisher Scientific (1 mentions) Lsrfortessa x 20, by BD (1 mentions)

4 protocols using 70 μm cell strainer

Oropharyngeal Swab and Nasal Lavage Processing

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Swabs were collected from the distal oropharynx and immediately placed in 2 mL of complete tissue culture medium in 15 mL polypropylene tubes on ice. The samples were lightly vortexed to free cells from the swabs. Fluid was transferred to a new tube, and 2 mL of PBS was added to the empty tube containing the swab, vortexed again, and pooled with the first sample. The sample then was filtered using a 70 μm cell strainer (Celltreat, Pepperell, Massachusetts) to remove debris and mucus. Samples were centrifuged, the cell pellets were resuspended in 0.5 mL of PBS, and the cells were counted.
Nasal lavage samples were filtered, washed, centrifuged, resuspended, and counted in the same fashion as for oropharyngeal swab samples. Cells were stained in the same manner as described for PBMC, and equal numbers were analyzed by flow cytometry to determine the cell type and activation.

Wheat W., Chow L., Coy J., Contreras E., Lappin M, & Dow S. (2019). Activation of upper respiratory tract mucosal innate immune responses in cats by liposomal toll‐like receptor ligand complexes delivered topically. Journal of Veterinary Internal Medicine, 33(2), 838-845.

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Comprehensive Organ Necropsies and CFU Quantification

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As previously described, the mice were euthanized by CO₂ and cardiac puncture [18 (link)]. Bladder, uterus, spleen, kidney, cecum, small intestine, stomach, liver, heart, lung, and brain necropsies were collected; weighed; and used for CFU counts and histology. For CFU counts, the organs were homogenized in Hanks Sodium Balanced Salt (HBSS) (Thermo Fisher, Waltham, MA, USA) through a 70 μm cell strainer (Cell Treat, Pepperell, MA, USA). A 1:100 dilution of organ homogenate was plated onto a Sabouraud dextrose agar plate with antibiotics (chloramphenicol (20 μg/mL), penicillin (20 U), gentamicin (40 μg/mL), and streptomycin (40 μg/mL)) [18 (link)].

Pichowicz A.M., Torres S.R., Torres-Velez F.J., Longyear A.D., Singh N., Lasek-Nesselquist E, & De Jesus M. (2022). Depletion of the Microbiota Has a Modest but Important Impact on the Fungal Burden of the Heart and Lungs during Early Systemic Candida auris Infection in Neutropenic Mice. Microorganisms, 10(2), 330.

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Nasal and oropharyngeal sampling for cell analysis

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Nasal lavage fluid was diluted with 3 ml of complete DMEM and oropharyngeal swabs were collected from the distal oropharynx and immediately placed in 2 ml of complete tissue culture medium in 15 ml polypropylene tubes on ice. To dislodge cells from the swabs, the samples were lightly vortexed. Fluid was subsequently transferred to a new tube and the swabs were again rinsed with 2 ml of PBS and pooled with the first wash. Pooled samples were filtered through a 70 μm cell strainer (CellTreat, Pepperell, MA) to remove large debris and mucus. Samples were then centrifuged and the pellets were resuspended in 0.5 ml of PBS and 10 μl aliquots were stained with 0.4% trypan blue and blue-excluding cells were counted using a Nexcelom Cellometer™ Auto T4 cell counter (Nexcelom; Lawrence, MA). Nasal lavage samples were processed similarly. To determine cell types and activation states, equivalent numbers of cells were processed and stained for flow cytometric analyses as described above.

Wheat W., Chow L., Kuzmik A., Soontararak S., Kurihara J., Lappin M, & Dow S. (2019). Local immune and microbiological responses to mucosal administration of a Liposome-TLR agonist immunotherapeutic in dogs. BMC Veterinary Research, 15, 330.

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Flow Cytometry Analysis of Immune Cells in Tissue Gels

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Gels were
surgically removed
from the animals after euthanasia. The gels were processed using mechanical
disruption to create a cell suspension that was then filtered through
a 70 μm cell strainer (Celltreat). To count total cells in the
gels, the single-cell suspensions were stained with anti-mouse CD45
and DAPI to count live leukocytes. The cells were mixed with CountBright
Absolute Counting beads (ThermoFisher) and analyzed on a LSRFortessa
X-20 flow cytometer (BD Biosciences). To analyze neutrophil and other
myeloid cell counts in the gels, the single-cell suspensions were
incubated with anti-CD16/CD32 (produced in house from 2.4G2 hybridoma)
to block Fc receptor binding and then stained with anti-mouse CD45,
CD3, CD19, CD11c, Ly6G, CD11b, and MHCII in FACS buffer (2 mM EDTA,
2% FBS in PBS) for 30 min on ice. Dead cells were excluded by DAPI
staining. Cells were acquired on a LSRFortessa X-20, and fcs files
were analyzed using FlowJo 10 (FlowJo LLC). See SI Figure 13 for the gating strategy and SI Table 3 for the antibody panel.

Roth G.A., Gale E.C., Alcántara-Hernández M., Luo W., Axpe E., Verma R., Yin Q., Yu A.C., Lopez Hernandez H., Maikawa C.L., Smith A.A., Davis M.M., Pulendran B., Idoyaga J, & Appel E.A. (2020). Injectable Hydrogels for Sustained Codelivery of Subunit Vaccines Enhance Humoral Immunity. ACS Central Science, 6(10), 1800-1812.

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