Lymphocytes at a concentration of 2 × 106 cells/mL were incubated with 50 µL of stimuli at 100 ng/mL and with 5µL of Fluo-3 AM solution at 1 mM (Sigma, St. Louis, MO, USA) for 2 h at 37 °C [6 (link),7 (link),62 (link)]. Cells were washed and resuspended in Hank’s Balanced Salt Solution (HBSS) with bovine serum albumin (BSA). Fluoroskan Ascent FL® was used to measure fluorescence intensity with 485 nm excitation and 538 nm emission filters, and the results were described according to fluorescence intensity.
Pereira G.D., Morais T.C., França E.L., Daboin B.E., Bezerra I.M., Pessoa R.S., de Quental O.B., Honório-França A.C, & de Abreu L.C. (2023). Leptin, Adiponectin, and Melatonin Modulate Colostrum Lymphocytes in Mothers with Obesity. International Journal of Molecular Sciences, 24(3), 2662.
Corresponding Organization : Universidade Federal de Mato Grosso
Other organizations :
Universidade de São Paulo, Escola Superior de Ciências da Santa Casa de Misericórdia de Vitória, Universidade Federal do Espírito Santo, University of Limerick, Faculdade de Medicina do ABC
Incubation of lymphocytes with 50 µL of stimuli at 100 ng/mL
dependent variables
Fluorescence intensity measured using Fluoroskan Ascent FL® with 485 nm excitation and 538 nm emission filters
control variables
Lymphocyte concentration of 2 × 10^6 cells/mL
Incubation with 5 µL of Fluo-3 AM solution at 1 mM for 2 h at 37 °C
Washing and resuspension of cells in Hank's Balanced Salt Solution (HBSS) with bovine serum albumin (BSA)
controls
No positive or negative controls were explicitly mentioned in the input text.
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