2-COLOR FLOW CYTOMETRIC ANALYSIS OF INTRAERYTHROCYTIC MALARIA PARASITE DNA AND SURFACE MEMBRANE-ASSOCIATED ANTIGEN IN ERYTHROCYTES INFECTEDWITH PLASMODIUM-FALCIPARUM
K. Pattanapanyasat et al., 2-COLOR FLOW CYTOMETRIC ANALYSIS OF INTRAERYTHROCYTIC MALARIA PARASITE DNA AND SURFACE MEMBRANE-ASSOCIATED ANTIGEN IN ERYTHROCYTES INFECTEDWITH PLASMODIUM-FALCIPARUM, Cytometry, 14(4), 1993, pp. 449-454
Citations number
26
Categorie Soggetti
Cytology & Histology","Biochemical Research Methods
A method for fixation of Plasmodium falciparum infected erythrocytes a
nd solubilization of the erythrocyte membrane with detergent was devel
oped. This method was applied to two-color flow cytometric analysis of
both intraerythrocytic (IE) malaria DNA and parasite-derived antigen
on the erythrocyte surface membrane. Infected erythrocytes were fixed
with 0.025% glutaraldehyde followed by treatment with 1% saponin to ga
in access to intramembranous components and allow antibody to interact
with antigen. DNA of IE parasite was subsequently stained with propid
ium iodide. Using this procedure cell morphology was well preserved wi
th excellent parasite DNA staining. Using anti-malaria antibodies whic
h recognize ring-infected erythrocyte surface antigen (Pf155/RESA), we
observed that glutaraldehyde-fixed saponin treated infected erythrocy
tes exhibited a variable immunofluorescence intensity as assessed by b
oth flow cytometry and fluorescence microscopy. Ring-infected cells di
splayed strong immunofluorescence staining, whereas a weak signal was
detected on cells containing schizonts. Simultaneous measurement of pa
rasite DNA and antigen in the infected erythrocyte membrane can facili
tate the study of antigen expression in the cell membrane in associati
on with development of IE parasites.