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Multiparameter Flow Cytometry Best Practices

Multiparameter flow cytometry is an important tool for biomedical scientists, especially for preclinical drug and vaccine research. It is an invaluable tool for detailed analysis of receptors, signaling, and effector molecules as well as nucleic acids in individual cells and cell populations. Technological advancements have made it possible to design and execute flow cytometry studies that simultaneously evaluate multiple parameters. However, evaluating multiple parameters at the cellular level is not an easy task.  Below are some sample preparation, panel design, and data analysis aspects for new and inexperienced flow cytometry users to consider.

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Flow Cytometry: A Vital Tool for Evaluating Vaccine Effectiveness

Flow cytometry uses light scattering, light excitation and emission of fluorochrome molecules to gather detailed information about specific cells or particles of interest. Using antibody-fluorophore conjugates flow cytometry allows researchers to draw phenotype-dependent conclusions about cellular phenotype and function.  The fluorophores present on bound antibodies excited by lasers yield detailed…

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Fig. 1 Survival after challenge with INFV H1N1 A/Pert/261/2009 (Tamiflu-resistant strain). Inoculum 1xLD90=1.0E+05 PFU/mouse
Survival after challenge with INFV H1N1 A/Pert/261/2009 (Tamiflu-resistant strain) 1.0E+05 PFU/mouse
Survival and weight change in BALB/c mice challenged with INFV A/ Texas/36/91 (H1N1) and treated with antiviral Osletamivir Phosphate (Tamiflu)
Lung viral load and Survival (30 % weight loss cut-off) in BALB/c mice challenged with INFV H3N2 A/HK/1/68.

Alpha (UK) – B. 1.1.7 / 501Y.V1

amino acid mutations: del69–70 HV, del144 Y, N501Y, A570D, D614G, P681H, T761I, S982A, D1118H

Beta (South Africa) – B.1.351

amino acid mutations: K417N, E484K, N501Y, D614G, A701V

Gamma (Brazil) – P.1

amino acid mutations: L18F, T20N, P26S, D138Y, R190S, K417T, E484K, N501Y, D614G, H655Y, T1027I

Epsilon (Ca, USA) B.1.427

amino acid mutations: L452R, D614G

SARS-CoV-2 Parental Strain Wild Type (Wuhan)
SARS-CoV-2 D614G Variant

amino acid mutations: D614G

Epsilon (Ca, USA) B.1.429

amino acid mutations: S13I, W152C, L452R, D614G

SARS-CoV-2 Delta Variant

amino acid mutations: L452R, E484Q