Repetitive cell 'jumps' during hypotonic lysis of erythrocytes observed with a simple flow chamber

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Abstract

Summary

This is the first report describing cell movements of a repetitive character during hypotonic lysis of erythrocytes (haemolysis). A new, simply constructed chamber is described for microscopy of freely suspended cells, for example, blood cells, during the inflow of a new medium. Hypotonic haemolysis of individual red blood cells was studied. During the first phase of haemolysis discontinuities were found: the cells made between zero and seven sudden movements or 'jumps', interpreted as caused by an ejection of cytoplasm due to excess intracellular hydraulic pressure and the formation of a hole. After pressure equilibration the hole resealed spontaneously. When, after one or two jumps, the inflow of hypotonic medium was stopped, the haemolytic process was interrupted but continued after restarting the flow. Inhibition of haemoglobin (Hb) release by 80% by external Ficoll® did not affect the number of 'jumps'. Since the optical contrast was reduced owing to Hb release after the last jump, less than 20% of the Hb loss can be associated with the jumps. Ejections of faint clouds of Hb were observed mainly in the presence of Ficoll®, but only after the last jump.

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