Ghost formation in post-lytic phase of colloid-osmotic lysis of human erythrocytes

Lysis of human red blood cells induced by hypotonic shock, non-ionic detergent (triton X-100), and pore-formers (a polyene antibiotics, amphotericin B, a bee venom toxin, melittin, and alpha-hemolysin from Staphylococcus aures) was studied using photometry, electrical detection of vesicular structures, and light microscopy. Hypotonic shock and in non-ionic detergent triton X-100 in high concentrations were found to produce unsealed, electrically transparent membrane fragments. Return to normal tonicity after hypotonic shock allowed membrane fragments to reseal as it was detected by electric conductivity. Meanwhile, when the cells were lysed by pore-forming agents, the total number of sealed membrane formations detected electronically after complete release of hemoglobin was equal to that of intact cells before treatment. Therefore, colloid-osmotic lysis by channel-forming substances leaded to the formation of electrically detectable "ghosts" in the whole concentration range studied. These formations were visualized and characterized by light microscopy. Following lysis, the "ghosts" displayed remarkable shrinkage. The average diameter of the 'ghosts' was 11.6 +/- 2.9 mum in early post-lytic phase, and decreased down to 7.9 +/- 3.3 mum within 60 min after complete release of hemoglobin.