Measurement of human red blood cell deformability using a single micropore on a thin Si3N4 film

The filtration method for the evaluation of the RBC deformability has been further refined to simulate the deformations encountered in the recticuloendothelial system (in particular the spleen), a recognized site of aged and sickled cells removal. The core of the developed measuring system is a very thin (0.4 micron thick) filter that consists of single micropore (diameters down to 1 micron) on a Si3N4 film which has been constructed using silicon microfabrication techniques. Individual RBC's deformability is quantified measuring the cell pore passage time. From one blood sample 200 passage times are analyzed by a computer, displaying mean and median values as deformability indexes, and class and cumulative histograms for studying the passage times distribution. In this paper the effectiveness of the developed system as a routine clinical evaluation tool is demonstrated by studying several factors that are known to affect the RBC deformability, such as temperature, addition of diamide and glutaraldehyde, and blood storage conditions. In addition, it is experimentally demonstrated that the human RBC can traverse a pore with a diameter as small as 1 micron when the pore length is very short, thus broadening the experimental conditions under which the RBC deformability (fluidity) can be studied.