Sample preparation
This protocol is extracted from research article:
Rapid and inefficient kinetics of sickle hemoglobin fiber growth
Sci Adv, Mar 13, 2019; DOI: 10.1126/sciadv.aau1086

Sickle hemoglobin (H0392; Sigma-Aldrich, St. Louis, MO) samples were suspended in 0.1 M potassium phosphate buffer (pH 7.0). Before imaging, HbS samples were deoxygenated on ice under a vacuum for 1 hour, after which samples were then brought to the final imaging concentration by further dilution in potassium phosphate buffer with sodium metabisulfite (final concentration of 50 mM) as an oxygen scavenger. Samples were placed on glass slides, covered with an acid-cleaned coverslip (6), and then sealed with CoverGrip sealant (Biotium, Fremont, CA). All samples were prepared at 4°C. Polymerization was initialized by temperature jump upon placing the slide on the microscope stage, where temperature was maintained at 25° or 37°C by use of an airstream incubator (Nevtek, Burnsville, VA) in combination with an objective heater (OkoLab SRL, Pozzuoli, Italy). Three independent sample preparations were performed for each temperature.

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