X50 symphony flow cytometer

Manufactured by BD

The X50 Symphony flow cytometer is a high-performance, multi-parameter cell analysis instrument. It is designed to accurately measure and analyze the physical and fluorescent characteristics of cells or other particles in a fluid sample.

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Lab products found in correlation

Facsaria, by BD (1 mentions) Gentlemacs, by Miltenyi Biotec (1 mentions) Lung cell isolation buffer, by Miltenyi Biotec (1 mentions)

Spelling variants of this product

Symphony flow cytometer (68 citations) Symphony cytometer (17 citations)

2 protocols using x50 symphony flow cytometer

Isolation and Purification of Lung Immune Cells

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Lungs were digested and dissociated via GentleMACS and Lung Cell Isolation Buffer (Miltenyi Biotec, CA). Digested lung was passged through a 100 μm cell strainer and an aliquot removed for CFU. Cells were washed and purified with 37% Percoll. Cells for sorting, were washed, counted and subsequently surface stained in a BSL3 containment area under sterile conditions. The following cell populations were sorted to 90–97% purity, CD45.1⁺ (WT) or CD45.1^– (Il1r1^−/−) CD11b⁺ cells, CD11b⁺ Gr1^high (neutrophils), CD11b⁺ Gr1^low (myeloid) and plated for CFU counts. Antibodies against I-A^b (clone M5/114.15.2), Ly6G (1A8), CD11c (HL3 and N418), CD45.1 (A20), CD45.2 (104), TCRβ (H57–597), NK1.1 (PK136), CD11b (M1/70), CD45 (30-F11), Gr-1 (RB6, 8C5) and Fixable live/dead Cell Stain were obtained from eBioscience/Thermofisher, Biolegend or BD Pharmingen. Samples were acquired on a X50 Symphony flow cytometer or sorted on a FACS Aria (BD Biosciences, San Jose, CA) and analyzed using FlowJo software.

Bohrer A.C., Tocheny C., Assmann M., Ganusov V.V, & Mayer-Barber K.D. (2018). IL-1R1 mediates host resistance to Mycobacterium tuberculosis by trans-protection of infected cells. Journal of immunology (Baltimore, Md. : 1950), 201(6), 1645-1650.

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Tracking Mycobacterium tuberculosis in Lungs

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For in vivo infections, WT B6.SJL (CD45.1/1) mice were intrapharyngeally inoculated with 50 CFU of Mtb HN878 (WT, L5 attB:P_left^*mWasabi, L5 attB:P_left^*mCherry, or L5 attB:P_left^*mScarlet). Lungs were harvested 37 days post-infection and dissociated via GentleMACS and Lung Cell Isolation Buffer (Miltenyi Biotec). Digested lungs were passed through a 100 μm cell strainer and cells washed and purified with 37% Percoll/RPMI. Cells for flow cytometry were washed, counted and subsequently fixed overnight using an Intracellular Fixation & Permeabilization Buffer (eBioscience/Thermo Fisher Scientific). Samples were acquired on a X50 Symphony flow cytometer (BD Biosciences) and analyzed using FlowJo software (BD Biosciences).

Kolbe K., Bell A.C., Prosser G.A., Assmann M., Yang H.J., Forbes H.E., Gallucci S., Mayer-Barber K.D., Boshoff H.I, & Barry III C.E. (2020). Development and Optimization of Chromosomally-Integrated Fluorescent Mycobacterium tuberculosis Reporter Constructs. Frontiers in Microbiology, 11, 591866.

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