flow cytometry tube with strainer cap  (Sony)

Journal: Cell Communication and Signaling : CCS

Article Title: F0F1 ATP synthase regulates extracellular calcium influx in human neutrophils by interacting with Ca v 2.3 and modulates neutrophil accumulation in the lipopolysaccharide-challenged lung

doi: 10.1186/s12964-020-0515-3

Figure Lengend Snippet: F-ATPase/Ca v 2.3 functional complexes regulate ERK 1/2 phosphorylation and ROS production after fMLP activation of neutrophils. a. A representative western blot is presented to show the effect of F-ATPase/Ca v 2.3 functional complexes on ERK 1/2 phosphorylation. Neutrophils were pretreated with 400 nM SNX482 and 50 μg/ml oligomycin A in Ca 2+ -containing HBSS for 30 min, followed by 1 min of 100 nM fMLP activation. Cells in Ca 2+ -containing HBSS or Ca 2+ -free HBSS incubated with or without fMLP were used as positive or negative controls, respectively. b. The effect of F-ATPase/Ca v 2.3 functional complexes on ROS production was assessed by flow cytometry. DHR123-loaded neutrophils were pretreated with 400 nM SNX482 and 50 μg/ml oligomycin A in Ca 2+ -containing HBSS for 30 min, followed by 30 min of 100 nM fMLP incubation. Cells in Ca 2+ -containing HBSS or Ca 2+ -free HBSS incubated with or without fMLP were used as positive or negative controls, respectively

Article Snippet: The aggregated cells were removed by filtration through a 35-μm cell strainer snap cap on a flow cytometry tube (Falcon, Corning, Cambridge, MA).

Techniques: Functional Assay, Activation Assay, Western Blot, Incubation, Flow Cytometry

Journal: Andrology

Article Title: A detailed protocol for a rapid analysis of testicular cell populations using flow cytometry

doi: 10.1111/andr.12066

Figure Lengend Snippet: Distribution of cells according to DNA amount in the ontogenical series of rat testis flow cytometry. The population dynamics of cells treated according to the present protocol and analyzed with propidium iodide ( PI ) recapitulate rat testicular ontogenesis. Diploid (2C), tetraploid (4C) and haploid (1C) were gated using the FSC / PI density plot and PI histogram. N = 5/group.

Article Snippet: A flow cytometry tube cell strainer cap (cat. # 352235; BD Falcon, BD Biosciences, Franklin Lakes, NJ, USA) attached to an Eppendorf tube functions well for the filtration.

Techniques: Flow Cytometry

Journal: Andrology

Article Title: A detailed protocol for a rapid analysis of testicular cell populations using flow cytometry

doi: 10.1111/andr.12066

Figure Lengend Snippet: Flow cytometry analysis using vimentin antibody recapitulates somatic cell dynamics during testis development in the rat. (A) Immunofluorescence on paraformaldehyde‐fixed paraffin‐embedded adult rat testis using anti‐vimentin‐Alexa488 (green) and DAPI as a nuclear counter stain. Solid arrowheads = Sertoli cells, open arrowheads = interstitial cells. Scale bar represents 50 μm. (B) Vimentin‐positive cells (black) clustering within the 2C cell population (white) in 5, 10, 16, 24 days and adult rat testis. N = 5/group. C. Vimentin‐positive cells (black) clustering within the proliferating cell population (white) in 5, 10, 16, 24 days and adult rat testis. (D) Representative histograms of propidium iodide and vimentin staining from 5‐, 10‐, 16‐, 24‐day‐old and adult. The proportion of proliferating vimentin‐positive cells (black histogram) was high in 5‐day‐old testis (C and D), but their proportion gradually decreased so that they are absent from adult testes. The majority of the observed 2C cells in the 5‐day‐old testis are vimentin‐ positive(B and D). As spermatogenesis progresses and the number of spermatogonia increases, the relative amount of vimentin‐positive cells in the 2C population decreases (B and D).

Article Snippet: A flow cytometry tube cell strainer cap (cat. # 352235; BD Falcon, BD Biosciences, Franklin Lakes, NJ, USA) attached to an Eppendorf tube functions well for the filtration.

Techniques: Flow Cytometry, Immunofluorescence, Staining

Journal: Andrology

Article Title: A detailed protocol for a rapid analysis of testicular cell populations using flow cytometry

doi: 10.1111/andr.12066

Figure Lengend Snippet: γH2AX‐S139 can be used as a marker for specific germ cell subpopulations in the flow cytometry assay. (A) Immunofluorescence on paraformaldehyde‐fixed paraffin‐embedded 16dpp and adult rat testis using anti‐γH2AX‐S139 (red) and DAPI as a nuclear counter stain. Double arrowhead = 4C cells with γH2AX‐positive sex body. Open arrowhead = γH2AX ‐positive spermatogonia. Scale bar represents 50 μm. (B–D) In the adult testis, γH2AX‐S139‐positive cell are detected in the subpopulations expected based on literature: Fraction of 1C and 2C cells and the majority of 4C cells. No γH2AX‐S139‐positive cells are observed at 16 dpp in the 1C cells population. (B) Representative histogram of 16 dpp rat testis stained with γH2AX‐S139‐antibody (black) and DNA stain (FxCycle, white). (C) Representative histogram of adult rat testis stained with γH2AX‐S139‐antibody (black) and DNA stain (FxCycle, white). (D) γH2AX‐S139‐positive cells (black) clustering within the 2C cell population (white) in 16‐day‐old and adult rat testis. N = 5/group. (E) γH2AX‐S139‐positive cells (black) clustering within the 4C cell population (white) in 16‐day‐old and adult rat testis. N = 5/group. (F) γH2AX‐S139‐positive cells (black) clustering within the 1C cell population (white) in 16‐day‐old and adult rat testis. N = 5/group.

Article Snippet: A flow cytometry tube cell strainer cap (cat. # 352235; BD Falcon, BD Biosciences, Franklin Lakes, NJ, USA) attached to an Eppendorf tube functions well for the filtration.

Techniques: Marker, Flow Cytometry, Immunofluorescence, Staining