A non-woven polymeric matrix for separating leukocytes from a blood sample includes a non-woven three dimensional matrix formed from polymeric fibers having a predetermined pore volume fraction including a defined channel configuration, a predetermined pore size in the range of from about 10 .mu.m to about 250 .mu.m, and a plurality of connections between the plurality of fibers. The matrix is configured so as to remove at least about 98% of leukocytes from at least one unit of packed red blood cells. Methods of making and using the matrix are also provided.