Publication: Flow cytometric two‐color staining technique for simultaneous determination of human erythrocyte membrane antigen and intracellular malarial DNA
Issued Date
1992-01-01
Resource Type
ISSN
10970320
01964763
01964763
Other identifier(s)
2-s2.0-0026529059
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Mahidol University
Rights Holder(s)
SCOPUS
Bibliographic Citation
Cytometry. Vol.13, No.2 (1992), 182-187
Suggested Citation
Kovit Pattanapanyasat, H. Kyle Webster, Rachanee Udomsangpetch, Wanchai Wanachiwanawin, Kosol Yongvanitchit Flow cytometric two‐color staining technique for simultaneous determination of human erythrocyte membrane antigen and intracellular malarial DNA. Cytometry. Vol.13, No.2 (1992), 182-187. doi:10.1002/cyto.990130212 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/22274
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Title
Flow cytometric two‐color staining technique for simultaneous determination of human erythrocyte membrane antigen and intracellular malarial DNA
Abstract
A novel fixative and permeabilization method is described which allows simultaneous flow cytometric detection of red blood cell membrane antigen and intracellular malaria parasites. To illustrate the method, red blood cells from patients with paroxysmal nocturnal bemoglobinuria were infected with Plasmodium falciparum and maintained in synchronous red blood cell culture. The infected red blood cells were immunolabeled with antibodies directed to the complement regulatory protein decay‐accelerating factor (DAF) followed by subsequent fixations in paraformaldehyde and then glutaraldehyde in phosphate‐buffered saline. Finally, DNA of the intraerythrocytic parasites was stained with propidium iodide. Using this technique, cellular morphology was well preserved, no cell aggregation was observed, and high‐quality indirect immunofluorescence and parasite DNA staining were obtained with negligible nonspecific labelling. Simultaneous measurement of parasite DNA and red blood cell membrane determinants makes possible the investigation of alterations of red cell membrane proteins in association with development of intracellular malaria parasites. Copyright © 1992 Wiley‐Liss, Inc.