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UNIT 11.20 Cytometry in Malaria—A Practical Replacement for Microscopy?

  1. Howard M. Shapiro1,
  2. Simon H. Apte2,
  3. Grace M. Chojnowski2,
  4. Thomas Hänscheid3,
  5. Maria Rebelo3,
  6. Brian T. Grimberg4

Published Online: 1 JUL 2013

DOI: 10.1002/0471142956.cy1120s65

Current Protocols in Cytometry

Current Protocols in Cytometry

How to Cite

Shapiro, H. M., Apte, S. H., Chojnowski, G. M., Hänscheid, T., Rebelo, M. and Grimberg, B. T. 2013. Cytometry in Malaria—A Practical Replacement for Microscopy?. Current Protocols in Cytometry. 65:11.20.1–11.20.23.

Author Information

  1. 1

    The Center for Microbial Cytometry, West Newton, Massachusetts

  2. 2

    Queensland Institute of Medical Research, Brisbane, Queensland, Australia

  3. 3

    Instituto de Medicina Molecular, Faculdade de Medicina da Universidade de Lisboa, Lisbon, Portugal

  4. 4

    Center for Global Health and Diseases, Case Western Reserve University School of Medicine, Cleveland, Ohio

Publication History

  1. Published Online: 1 JUL 2013
  2. Published Print: JUL 2013

Literature Cited

  1. Literature Cited
  • Apte, S.H., Groves, P.L., Roddick, J.S., P. da Hora, V., and Doolan, D.L. 2011. High-throughput multi-parameter flow-cytometric analysis from micro-quantities of plasmodium-infected blood. Int. J. Parasitol. 41:1285-1294.
  • Barber, B.E., William, T., Grigg, M.J., Yeo, T.W., and Anstey, N.M. 2013. Limitations of microscopy to differentiate Plasmodium species in a region co-endemic for Plasmodium falciparum, Plasmodium vivax and Plasmodium knowlesi. Malar. J. 12:8.
  • Basco, L.K. 2007. Field Application of In Vitro Assays for the Sensitivity of Human Malaria Parasites to Antimalarial Drugs. World Health Organization, Geneva.
  • Boissière, A., Arnathau, C., Duperray, C., Berry, L., Lachaud, L., Renaud, F., Durand, P., and Prugnolle, F. 2012. Isolation of Plasmodium falciparum by flow-cytometry: Implications for single-trophozoite genotyping and parasite DNA purification for whole-genome high-throughput sequencing of archival samples. Malar. J. 11:163.
  • Bosch, F. and Rosich, L. 2008. The contributions of Paul Ehrlich to pharmacology: A tribute on the occasion of the centenary of his Nobel Prize. Pharmacology 82:171-179.
  • Campo, J.J., Aponte, J.J., Nhabomba, A.J., Sacarlal, J., Angulo-Barturen, I., Jiménez-Díaz, M.B., Alonso, P.L., and Dobaño, C. 2011. Feasibility of flow cytometry for measurements of Plasmodium falciparum parasite burden in studies in areas of malaria endemicity by use of bidimensional assessment of YOYO-1 and autofluorescence. J. Clin. Microbiol. 49:968-974.
  • Ch'ng, J.-H., Kotturi, S.R., Chong, A. G-L., Lear, M.J., and Tan, K.S.-W. 2010. A programmed cell death pathway in the malaria parasite Plasmodium falciparum has general features of mammalian apoptosis but is mediated by clan CA cysteine proteases. Cell Death Dis. 1:e26.
  • Clark, G., and Kasten, F.H. 1983. History of Staining, 3rd ed. Williams & Wilkins, Baltimore, Maryland.
  • Clark, M., Fisher, N.C., Kasthuri, R., and Cerami Hand, C. 2013. Parasite maturation and host serum iron influence the labile iron pool of erythrocyte stage Plasmodium falciparum. Br. J. Haematol. 161:262-269.
  • Cox-Singh, J. 2012. Zoonotic malaria: Plasmodium knowlesi, an emerging pathogen. Curr. Opin. Infect. Dis. 25:530-536.
  • Dame, J.B. and McCutchan, T.F. 1987. Plasmodium falciparum: Hoechst dye 33258-CsCl ultracentrifugation for separating parasite and host DNAs. Exp. Parasitol. 64:264-266.
  • De Kruif, P. 1926. Microbe Hunters (republished 1996). Harcourt, San Diego.
  • Desowitz, R.S. 1991. The Malaria Capers: Tales of Parasites and People. Norton, New York.
  • Frita, R., Rebelo, M., Pamplona, A., Vigario, A.M., Mota, M.M., Grobusch, M.P., and Hänscheid, T. 2011. Simple flow cytometric detection of haemozoin containing leukocytes and erythrocytes for research on diagnosis, immunology and drug sensitivity testing. Malar. J. 10:74.
  • Frita, R., Carapau, D., Mota, M.M., and Hänscheid, T. 2012. In vivo hemozoin kinetics after clearance of Plasmodium berghei infection in mice. Malar. Res. Treat. 2012:373086.
  • Fuehrer, H.-P., Stadler, M.-T., Buczolich, K., Blöeschl, I., and Noedl, H. 2012. Two techniques for simultaneous identification of Plasmodium ovale curtisi and Plasmodium ovale wallikeri by use of the small-subunit rRNA gene. J. Clin. Microbiol. 50:4100-4102.
  • Garcia, L.S. 2007. Diagnostic Medical Parasitology, 5th ed. ASM Press, Washington, D.C.
  • Garfield, S. 2001. Mauve. Norton, New York.
  • Gaynes, R.P. 2011. Germ Theory: Medical Pioneers in Infectious Diseases. ASM Press, Washington, D.C.
  • Gerena, Y., Gonzalez-Pons, M., and Serrano, A.E. 2011. Cytofluorometric detection of rodent malaria parasites using red-excited fluorescent dyes. Cytometry A 79:965-972.
  • Grimberg, B.T. 2011. Methodology and application of flow cytometry for investigation of human malaria parasites. J. Immunol. Methods 367:1-16.
  • Grimberg, B.T., Erickson, J.J., Sramkoski, R.M., Jacobberger, J.W., and Zimmerman, P.A. 2008. Monitoring Plasmodium falciparum growth and development by UV flow cytometry using an optimized Hoechst-thiazole orange staining strategy. Cytometry A 73:546-554.
  • Grimberg, B.T., Jaworska, M.M., Hough, L.B., Zimmerman, P.A., and Phillips, J.G. 2009. Addressing the malaria drug resistance challenge using flow cytometry to discover new antimalarials. Bioorg. Med. Chem. Lett. 19:5452-5457.
  • Grobusch, M.P., Hänscheid, T., Krämer, B., Neukammer, J., May, J., Seybold, J., Kun, J.F.J., and Suttorp, N. 2003. Sensitivity of hemozoin detection by automated flow cytometry in non- and semi-immune malaria patients. Cytometry B 55:46-51.
  • Gross, H.-J., Verwer, B., Houck, D., Hoffman, R.A., and Recktenwald, D. 1995. Model study detecting breast cancer cells in peripheral blood mononuclear cells at frequencies as low as 10-7. Proc. Natl. Acad. Sci. U.S.A. 92:537-541.
  • Hänscheid, T., Egan, T.J., and Grobusch, M.P. 2007. Haemozoin: From melatonin pigment to drug target, diagnostic tool, and immune modulator. Lancet Infect. Dis. 7:675-685.
  • Hänscheid, T., Romão, R., Grobusch, M.P., Amaral, T., and Melo-Cristino, J. 2011. Limitation of malaria diagnosis with the Cell-Dyn® analyser: Not all haemozoin-containing monocytes are detected or shown. Int. J. Lab. Hematol. 33:e14-16.
  • Hare, J.D. 1986. Two-color flow-cytometric analysis of the growth cycle of Plasmodium falciparum in vitro: Identification of cell cycle compartments. J. Histochem. Cytochem. 34:1651-1658.
  • Hare, J.D. and Bahler, D.W. 1986. Analysis of Plasmodium falciparum growth in culture using acridine orange and flow cytometry. J. Histochem. Cytochem. 34:215-220.
  • Hawass, Z., Gad, Y.Z., Ismail, S., Khairat, R., Fathalla, D., Hasan, N., Ahmed, A., Elleithy, H., Ball, M., Gaballah, F., Wasef, S., Fateen, M., Amer, H., Gostner, P., Selim, A., Zink, A., and Pusch, C.M. 2010. Ancestry and pathology in King Tutankhamun's family. JAMA 303:638-647.
  • Izumiyama, S., Omura, M., Takasaki, T., Ohmae, H., and Asahi, H. 2009. Plasmodium falciparum: Development and validation of a measure of intraerythrocytic growth using SYBR Green I in a flow cytometer. Exp. Parasitol. 121:144-150.
  • Janse, C.J., van Vianen, P.H., Tanke, H.J., Mons, B., Ponnudurai, T., and Overdulve, J.P. 1987. Plasmodium species: Flow cytometry and microfluorometry assessments of DNA content and synthesis. Exp. Parasitol. 64:88-94.
  • Janse, C.J., Ponnudurai, T., Lensen, A.H.W., Meuwissen, J.H.E.Th., Ramesar, J., Van der Ploeg, M., and Overdulve, J.P. 1988. DNA synthesis in gametocytes of Plasmodium falciparum. Parasitology 96:1-7.
  • Jogdand, P.S., Singh, S.K., Christiansen, M., Dziegiel, M.H., Singh, S., and Theisen, M. 2012. Flow cytometric readout based on Mitotracker Red CMXRos staining of live asexual blood stage malarial parasites reliably assesses antibody dependent cellular inhibition. Malar. J. 11:235.
  • Jouin, H., Goguet de la Salmonière, Y.O., Behr, C., Huyin Qan Dat, M., Michel, J.C., Sarthou, J.L., Pereira da Silva, L., and Dubois, P. 1995. Flow cytometry detection of surface antigens on fresh, unfixed red blood cells infected by Plasmodium falciparum. J. Immunol. Methods 179:1-12.
  • Jouin, H., Daher, W., Khalife, J., Ricard, I., Puijalon, O.M., Capron, M., and Dive, D. 2004. Double staining of Plasmodium falciparum nucleic acids with hydroethidine and thiazole orange for cell cycle stage analysis by flow cytometry. Cytometry A 57:34-38.
  • Jun, G., Lee, J.-S., Jung, Y.-J., and Park, J.-W. 2012. Quantitative determination of Plasmodium parasitemia by flow cytometry and microscopy. J. Korean Med. Sci. 27:1137-1142.
  • Karl, S., David, M., Moore, L., Grimberg, B.T., Michon, P., Mueller, I., Zborowski, M., and Zimmerman, P.A. 2008. Enhanced detection of gametocytes by magnetic deposition microscopy predicts higher potential for Plasmodium falciparum transmission. Malar. J. 7:66.
  • Karl, S., Davis, T.M.E., and St-Pierre, T.G. 2009. A comparison of the sensitivities of detection of Plasmodium falciparum gametocytes by magnetic fractionation, thick blood film microscopy, and RT-PCR. Malar. J. 8:98.
  • Kaushansky, A., Rezakhani, N., Mann, H., and Kappe, S.H.I. 2012. Development of a quantitative flow cytometry-based assay to assess infection by Plasmodium falciparum sporozoites. Mol. Biochem. Parasitol. 183:100-103.
  • Kim, C.C., Wilson, E.B., and DeRisi, J.L. 2010. Improved methods for magnetic purification of malaria parasites and haemozoin. Malar. J. 9:17.
  • Laveran, A. 1907. Protozoa as causes of diseases. Nobel lecture (http://www.nobelprize.org/).
  • Lawrence, C. and Olson, J.A. 1986. Birefringent hemozoin identifies malaria. Am. J. Clin. Pathol. 86:360-363.
  • Lewert, R.M. 1952. Nucleic acids in Plasmodia and the phosphorus partition of cells infected with Plasmodium gallinaceum. J. Infect. Dis. 91:125-144.
  • Malleret, B., Claser, C., Ong, A.S.M., Suwanarusk, R., Sriprawat, K., Howland, S.W., Russell, B., Nosten, F., and Rénia, L. 2011. A rapid and robust tri-color flow cytometry assay for monitoring malaria parasite development. Sci. Rep. 1:118.
  • Mandy, F., Nicholson, J., Autran, B., and Janossy, G. 2002. T-cell subset counting and the fight against AIDS: Reflections over a 20-year struggle. Cytometry 50:39-45.
  • Maude, R.J., Buapetch, W., and Silamut, K. 2009. A simplified, low-cost method for polarized light microscopy. Am. J. Trop. Med. Hyg. 81:782-783.
  • McMorrow, M.L., Aidoo, M., and Kachur, S.P. 2011. Malaria rapid diagnostic tests in elimination settings can they find the last parasite? Clin. Microbiol. Infect. 17:1624-1631.
  • Mendelow, B.V., Lyons, C., Nhlangothi, P., Tana, M., Munster, M., Wypkema, E., Liebowitz, L., Marshall, L., Scott, S., and Coetzer, T.L. 1999. Automated malaria detection by depolarization of laser light. Br. J. Haematol. 104:499-503.
  • Mühlpfordt, H. and Berger, J. 1989. Characterization and grouping of Trypanosoma brucei brucei, T.b. gambiense and T.b. rhodesiense by quantitative DNA-cytofluorometry and discriminant analysis. Trop. Med. Parasitol. 40:1-8.
  • Mühlpfordt, H. and Berger, J. 1990. Characterization and grouping of Trypanosoma cruzi stocks by DNA base-specific fluorochromes and discriminant analysis. Parasitol. Res. 76:319-325.
  • Mühlpfordt, H., Berger, J., and Glaser, N. 1985. Cytofluorometry as a method for the differentiation of trypanosomes. Trop. Med. Parasitol. 36:135-139.
  • Murray, C.J.L., Rosenfeld, L.C., Lim, S.S., Andrews, K.G., Foreman, K.J., Haring, D., Fullman, N., Naghavi, M., Lozano, R., and Lopez, A.D. 2012. Global malaria mortality between 1980 and 2010: A systematic analysis. Lancet 379:413-431.
  • Murray, C.K., Gasser, R.A. Jr., Magill, A.J., and Miller, R.S. 2008. Update on rapid diagnostic testing for malaria. Clin. Microbiol. Rev. 21:97-110.
  • O'Meara, W.P., Barcus, M., Wongsrichanalai, C., Muth, S., Maguire, J.D., Jordan, R.G., Prescott, W.R., and McKenzie, F.E. 2006. Reader technique as a source of variability in determining malaria parasite density by microscopy. Malar. J. 5:118.
  • O'Meara, W.P., Hall, B.F., and McKenzie, F.E. 2007. Malaria vaccine efficacy: The difficulty of detecting and diagnosing malaria. Malar. J. 6:36.
  • Ohrt, C., O'Meara, W.P., Remich, S., McEvoy, P., Ogutu, B., Mtalib, R., and Odera, J.S. 2008. Pilot assessment of the sensitivity of the malaria thin film. Malar. J. 7:22.
  • Pattanapanyasat, K., Udomsangpetch, R., and Webster, H.K. 1993. Two-color flow cytometric analysis of intraerythrocytic malaria parasite DNA and surface membrane-associated antigen in erythrocytes infected with Plasmodium falciparum. Cytometry 14:449-454.
  • Philipp, S., Oberg, H.-H., Janssen, O., Leippe, M., and Gelhaus, C. 2012. Isolation of erythrocytes infected with viable early stages of Plasmodium falciparum by flow cytometry. Cytometry A 81:1048-1054.
  • Rebelo, M., Shapiro, H.M., Amaral, T., Melo-Cristino, J., and Hänscheid, T. 2012. Haemozoin detection in infected erythrocytes for Plasmodium falciparum malaria diagnosis—prospects and limitations. Acta Trop. 123:58-61.
  • Rebelo, M., Sousa, C., Shapiro, H.M., Mota, M.M., Grobusch, M.P., and Hänscheid, T. 2013. A novel flow cytometric hemozoin detection assay for real-time sensitivity testing of Plasmodium falciparum. PLoS One 8:e61606.
  • Ross, R. 1903. An improved method for the microscopical diagnosis of intermittent fever. Lancet 161:86.
  • Ross, R. and Thomson, D. 1910. Some enumerative studies on malarial fever. Proc. Royal Soc. London B 83:159-173.
  • Shapiro, H.M. 1981. Flow cytometric estimation of DNA and RNA content in intact cells stained with Hoechst 33342 and pyronin Y. Cytometry 2:143-150.
  • Shapiro, H.M. 2003. Practical Flow Cytometry, 4th ed. Wiley-Liss, Hoboken, N.J.
  • Shapiro, H.M., Natale, P.J., and Kamentsky, L.A. 1979. Estimation of membrane potentials of individual lymphocytes by flow cytometry. Proc. Natl. Acad. Sci. U.S.A. 76:5728-5730.
  • Sinnis, P., De La Vega, P., Coppi, A., Krzych, U., and Mota, M.M. 2013. Quantification of sporozoite invasion, migration, and development by microscopy and flow cytometry. Methods Mol. Biol. 923:385-400.
  • Tilley, L., McFadden, G., Cowman, A., and Klonis, N. 2007. Illuminating Plasmodium falciparum-infected red blood cells. Trends Parasitol. 23:268-277.
  • Vorobjev, I.A., Buchholz, K., Prabhat, P., Ketman, K., Egan, E.S., Marti, M., Duraisingh, M.T., and Barteneva, N.S. 2012. Optimization of flow cytometric detection and cell sorting of transgenic Plasmodium parasites using interchangeable optical filters. Malar. J. 11:312.
  • Wongchotigul, V., Suwanna, N., Krudsood, S., Chindanond, D., Kano, S., Hanaoka, N., Akai, Y., Maekawa, Y., Nakayama, S., Kojima, S., and Looareesuwan, S. 2004. The use of flow cytometry as a diagnostic test for malaria parasites. Southeast Asian J. Trop. Med. Public Health 35:552-559.
  • World Health Organization. 2001. In vitro micro-test (mark III) for the assessment of the response of Plasmodium falciparum to chloroquine, mefloquine, quinine, amodiaquine, sufadoxine/pyrimethamine and artemisinin. World Health Organization, Geneva.
  • World Health Organization. 2008. Malaria Microscopy Quality Assurance Manual, Version 1. World Health Organization, Geneva.
  • World Health Organization. 2010. Parasitological confirmation of malaria diagnosis: Report of a WHO technical consultation Geneva, 6-8 October 2009. World Health Organization, Geneva.
  • World Health Organization. 2011. World Malaria Report 2011. World Health Organization, Geneva.