Cell Strainers

Content
1. What are cell strainers?
    1.1 Advantage of cell strainers
    1.2 Application of cell strainers
    1.3 Specification of cell strainers
    1.4 Specification of cell strainers
    1.5 Selection of cell strainers
2. Type of cell strainers
    2.1 Stainless-steel cell strainers
    2.2 Nylon cell strainers
3. Use of cell strainers
    3.1 Cleaning of cell strainers
4. Cell strainer for flow cytometry
5. How to buy cell strainers?

Cell strainers are generally used in laboratory cell culture, impurity filtration, cell dispersion, sample splitting, etc. The cell filter is usually made of a polypropylene frame, a special nylon screen combination. Common specifications are cell strainer 40μm, cell strainer 70μm and cell strainer 100μm screen apertures.

In cell biology research, whether it is for cell physiological characteristics, cell invasion, apoptosis, phagocytosis, and other physiological functions, the effect of environmental factors on cell marker expression, drug screening, and other applications, the prerequisite is to obtain the ideal homogenized single cell county fluid, so that subsequent experiments such as flow-through cell sorting (FACS) can be carried out smoothly and more accurate experimental results can be obtained. Cell strainers are the most common type of consumables used in cell experiments. It is the most common type of consumable used in cellular experiments and is used to filter out impurities in cellular experiments.

a. Three sizes available to match different types of cell samples: 40 µm, 70 µm, and 100 µm.

b. Three colors to distinguish different specifications: blue, white, and yellow.

c. Even distribution of mesh holes, providing consistent and reliable experimental results.

d. Top extended edge allows aseptic manipulation with surgical forceps.

e. Molded polypropylene frame can be color-coded for easy handling and differentiation.

f. For use with 50ml JET conical centrifuge tubes.

g. Gamma-irradiation sterilization.

h. No DNase/RNase, no heat source, no endotoxin.

The purpose of the cell strainers is to break up the cell mass during primary culture to obtain a single-cell suspension.

a. Single cell suspension obtained by filtration of blood cells isolated from bone marrow, pancreas, thymus, tonsils, and lymph nodes.

b. Preparation of samples for primary cell culture and immunization.

c. Filtration of gliadin from inactivated serum.

d. Preparation of frozen primary seeds.

The holes of cell strainers are punched by a special processing cell filter laser punching machine. The working principle is that the laser beam over the laser is highly concentrated in space and time, which can reduce the spot diameter to a micron level thus obtaining high power density, and can laser punch almost any material.

The advantage of laser perforation of cell filter is no burr, no sealing, no residue, and flat cut. In laser perforation of cell filter hole, perforation size is consistent, if there is a burr or uneven surface, the next process will be very troublesome, and laser processing makes the later processing faster and better quality.

Number of mesh of cell filter sieve × pore size = 15000 (choose the cell sieve with the corresponding mesh size according to the size of the required filter pore size of the experimental material).

Cell strainers are generally divided into two main materials, one is plastic products such as nylon, and the other is made of stainless steel.

Stainless-steel cell filter sieve is made of stainless steel, with a smooth surface, beautiful appearance, and durable features. Stainless-steel cell strainers have various specifications from 40 mesh to 800 mesh, which are generally used in laboratory cell culture, impurity filtration, cell dispersion, sample separation, etc. Stainless-steel cell strainers can be reused, generally using ultrasonic cleaning and then sterilization. The cell sieve is divided into two kinds of stainless-steel cell strainer with handle and stainless-steel cell strainer without handle, which is simple and convenient to operate for scientific research experiments.

Common flow test specimens can be used with 300-350 mesh nylon cell strainers, folded into a funnel shape, and used with hemostatic forceps.

a. Cell strainers cannot be soaked in acid after washing, they can be directly wet sterilized by wrapping them in tin foil. Do not dry grind when using. When filtering larger tissues, you need to add buffer while grinding.

b. The difference between 200 mesh and 300 mesh cell strainers is the size of the holes. This difference is not very big when doing flow type, but when using 300 mesh, be careful not to grind too much to avoid excessive connective tissue.

c. Rat lung tissue can be obtained after cell sieve grinding is a mixture of cells, including lung parenchymal cells, stromal cells, erythrocytes, and a small number of leukocytes, NK cells, and macrophages.

a. Disposable cell strainers are not recommended to be cleaned and disinfected, which may cause unevenness of the filter and affect the results of the next experiment.

b. Dilute alkali cleaning to remove the residual cells and tissues.

c. Soak and clean with descaling agent solution.

d. Tap water flushing, dH2O distilled water 3 times, ddH2O re-distilled water 3 times, packaging sterilization.

Flow cytometry is a technique for quantitative analysis and sorting of single cells or biological particles in suspension, which has the advantages of high speed, high precision, and accuracy, and is widely used in cell counting, cell sorting, molecular phenotype analysis, protein engineering and other research.

a. Remove the mouse spleen into a 15 ml centrifuge tube (without liquid) and place the tube on ice.

b. Add 5 ml of RBC lysis buffer to 15 ml of the centrifuge tube containing the spleen.

c. Pour RBC lysis buffer and spleen into a 10 cm cell culture dish.

d. Place the spleen on the rough side of a slide double-ended grinder (moistened with RBC lysis buffer), thoroughly crush the spleen in the slide double-ended grinder, and grind between the two-slide double-ended grinders until the spleen is dissociated.

e. Transfer the RBC lysate containing splenocytes to a 15 ml centrifuge tube.

f. Add 10 ml of medium to the centrifuge tube and invert 2-3 times.

g. Centrifuge at 1300 rpm at 4 °C for 00:05:00

h. Discard the supernatant and resuspend the precipitate in 3 ml of medium.

i. Filter the 70 μM cell strainer through a 50 ml centrifuge tube.

j. Dilute cells at 1:100 and count.

k. Dilute the single cell suspension to 1 x 107 cells/ml in RPMI medium (for flow assays, no cells need to be counted and counted).

l. Place splenocytes on ice until they enter the 96-well plate.

a. Prepare one RPMI medium containing Brefeldin A to a concentration of 10 μg/ml.

b. Vortex and spin all LPS stock solutions, then sonicate each one 00:10:00 . After sonication, vortex and spin again.

c. Prepare each stimulus (EC LPS, PA, YP, Lipid IVa) to twice its desired final concentration (2x) using medium containing Brefeldin A.

d. 100 μl of each stimulant is added to the appropriate wells of a 96-well plate 2.2 mL, including the unstimulated wells as controls.

e. Plate 100 μl of splenocytes into the appropriate wells (plate another set of splenocytes to be used as a single staining control).

f. Then incubate at 37 °C for 04:00:00.

a. After 04:00:00, treat the well plate with 20 μl of 20 mM EDTA.

b. Mix and incubate at 37 °C for 00:10:00.

c. Mix again after incubation, then centrifuge at 1300 rpm for 00:05:00.

d. Aspirate or flick the media.

e. Close all wells with 10 μl of diluted FcBlock (1:100 FcBloc in PBSA/az).

f. Incubate on ice at 00:15:00.

g. Add 100 μl PBSA/az and centrifuge at 1300 rpm for 00:05:00.

h. Aspirate or flick the media.

i. Resuspend/stain with 10 μl of antibody mixture: CD11b PerCP-Cy5.5, CD11c PE-Cy7, CD19 APC-Cy7 and CD3 APC (1:100Ab in PBSA/az) to the stimulated splenocyte sample.

j. Add 100 μl PBSA/az to the single staining control, followed by 1 μl of these single staining Ab: CD8 FITC, CD8 PE, CD4 PerCP-Cy5.5, CD8 PE-Cy7, CD19 APC-Cy7 and CD8 APC.

k. Incubate on ice protected from light (or wrapped in foil) at 00:20:00.

l. Add 100 μl PBSA/az, then centrifuge at 1300 rpm for 00:05:00.

m. Aspirate or flick the medium.

n. Resuspend in all wells with 100 μl BDcytofix/cytoperm solution.

o. Incubate on ice protected from light 00:20:00.

p. Add 100 μl of 1x BD Perm Wash to all wells.

q. Centrifuge at 1300 rpm for 00:05:00, then aspirate or flick the medium.

r. Resuspend/wash once with 200 μl 1x BD Perm Wash by pipetting up and down several times into all wells.

s. Centrifuge at 1300 rpm for 00:05:00, then aspirate or flick the medium.

t. Resuspend/stain the splenocyte samples with 50 μl of antibody mixture: TNF FITC and IL-12/IL-23p40 PE to the stimulated splenocytes.

u. Incubate on ice at 00:30:00 protected from light.

v. Add 150 μl 1xBD Perm wash to all wells and centrifuge at 1300 rpm for 00:05:00.

w. Aspirate or flick the media.

x. Resuspend/wash once with 200 μl 1x BD Perm Wash by pipetting up and down several times into all wells.

y. Centrifuge at 1300 rpm for 00:05:00, then aspirate or flick the medium.

z. Resuspend in a final volume of 200 μl of 1% paraformaldehyde solution (dilute 10% paraformaldehyde with 1xPBS).

aa. Store at 4 °C away from light (or wrap in foil) until ready for flow cytometry assay - transfer samples from 96-well plate to burette immediately prior to FACS.

If you are interested in our cell strainers or have any questions, please write an e-mail to info@antiteck.com, we will reply to you as soon as possible.