Ficoll, solution preparation

Ficoll dissolves slowly and the solutions should be prepared in advance. It is possible to freeze aliquots. 

Ficoll-Conray gradient is prepared as follows mix Conray 420 (33% [w/v] solution in water on the basis of sodium iothalamate content) to a 9% solution of Ficoll 400 in normal saline until the density of the mixture is 1.08 g/mL... 

During the dextran sedimentation, prepare the following in 50-mL conical tubes (1) 35 mL of 0.09% NaCl, (2) 20 mL of 1.7% NaCl, (3) 12 mL of Ficoll-Paque , and (4) 20 mL of injection- or irrigation-grade water. These solutions are normally stored at 4 °C, but to prevent priming and aggregation of PMNs, it is optimal to warm the solutions to room temperature. 

PMN and mononuclear leukocytes (MNL) were prepared from Ficoll-Hypaque density difference centrifugation as previously described by English (1976). After isolation, the cells were enumerated and diluted to desired concentrations in Hank s balanced salt solution (BBSS), Grand Island Biological Company, Grand Island, New York. 

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