{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,1]],"date-time":"2025-11-01T13:38:29Z","timestamp":1762004309376},"reference-count":34,"publisher":"Springer Science and Business Media LLC","issue":"1","content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Malar J"],"published-print":{"date-parts":[[2011,12]]},"abstract":"<jats:title>Abstract<\/jats:title>\n          <jats:sec>\n            <jats:title>Background<\/jats:title>\n            <jats:p>Malaria pigment (haemozoin, Hz) has been the focus of diverse research efforts. However, identification of Hz-containing leukocytes or parasitized erythrocytes is usually based on microscopy, with inherent limitations. Flow cytometric detection of depolarized Side-Scatter is more accurate and its adaptation to common bench top flow cytometers might allow several applications. These can range from the <jats:italic>ex-vivo<\/jats:italic> and <jats:italic>in-vitro<\/jats:italic> detection and functional analysis of Hz-containing leukocytes to the detection of parasitized Red-Blood-Cells (pRBCs) to assess antimalarial activity.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Methods<\/jats:title>\n            <jats:p>A standard benchtop flow cytometer was adapted to detect depolarized Side-Scatter. Synthetic and <jats:italic>Plasmodium falciparum<\/jats:italic> Hz were incubated with whole blood and PBMCs to detect Hz-containing leukocytes and CD16 expression on monocytes. C5BL\/6 mice were infected with <jats:italic>Plasmodium berghei<\/jats:italic> ANKA or <jats:italic>P. berghei<\/jats:italic> NK65 and Hz-containing leukocytes were analysed using CD11b and Gr1 expression. Parasitized RBC from infected mice were identified using anti-Ter119 and SYBR green I and were analysed for depolarized Side Scatter. A highly depolarizing RBC population was monitored in an <jats:italic>in-vitro<\/jats:italic> culture incubated with chloroquine or quinine.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Results<\/jats:title>\n            <jats:p>A flow cytometer can be easily adapted to detect depolarized Side-Scatter and thus, intracellular Hz. The detection and counting of Hz containing leukocytes in fresh human or mouse blood, as well as in leukocytes from <jats:italic>in-vitro<\/jats:italic> experiments was rapid and easy. Analysis of CD14\/CD16 and CD11b\/Gr1 monocyte expression in human or mouse blood, in a mixed populations of Hz-containing and non-containing monocytes, appears to show distinct patterns in both types of cells. Hz-containing pRBC and different maturation stages could be detected in blood from infected mice. The analysis of a highly depolarizing population that contained mature pRBC allowed to assess the effect of chloroquine and quinine after only 2 and 4 hours, respectively.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Conclusions<\/jats:title>\n            <jats:p>A simple modification of a flow cytometer allows for rapid and reliable detection and quantification of Hz-containing leukocytes and the analysis of differential surface marker expression in the same sample of Hz-containing <jats:italic>versus<\/jats:italic> non-Hz-containing leukocytes. Importantly, it distinguishes different maturation stages of parasitized RBC and may be the basis of a rapid no-added-reagent drug sensitivity assay.<\/jats:p>\n          <\/jats:sec>","DOI":"10.1186\/1475-2875-10-74","type":"journal-article","created":{"date-parts":[[2011,4,6]],"date-time":"2011-04-06T18:45:46Z","timestamp":1302115546000},"update-policy":"http:\/\/dx.doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":26,"title":["Simple flow cytometric detection of haemozoin containing leukocytes and erythrocytes for research on diagnosis, immunology and drug sensitivity testing"],"prefix":"10.1186","volume":"10","author":[{"given":"Rosangela","family":"Frita","sequence":"first","affiliation":[]},{"given":"Maria","family":"Rebelo","sequence":"additional","affiliation":[]},{"given":"Ana","family":"Pamplona","sequence":"additional","affiliation":[]},{"given":"Ana M","family":"Vigario","sequence":"additional","affiliation":[]},{"given":"Maria M","family":"Mota","sequence":"additional","affiliation":[]},{"given":"Martin P","family":"Grobusch","sequence":"additional","affiliation":[]},{"given":"Thomas","family":"H\u00e4nscheid","sequence":"additional","affiliation":[]}],"member":"297","published-online":{"date-parts":[[2011,3,31]]},"reference":[{"key":"1664_CR1","doi-asserted-by":"publisher","first-page":"675","DOI":"10.1016\/S1473-3099(07)70238-4","volume":"7","author":"T H\u00e4nscheid","year":"2007","unstructured":"H\u00e4nscheid T, Egan TJ, Grobusch MP: Hemozoin: from melatonin pigment to drug target, diagnostic tool, and immune modulator. Lancet Infect Dis. 2007, 7: 675-685.","journal-title":"Lancet Infect Dis"},{"key":"1664_CR2","doi-asserted-by":"publisher","first-page":"889","DOI":"10.1016\/j.micinf.2010.07.001","volume":"12","author":"MT Shio","year":"2010","unstructured":"Shio MT, Kassa FA, Bellemare MJ, Olivier M: Innate inflammatory response to the malarial pigment hemozoin. Microbes Infect. 2010, 12: 889-899. 10.1016\/j.micinf.2010.07.001.","journal-title":"Microbes Infect"},{"key":"1664_CR3","doi-asserted-by":"publisher","first-page":"142","DOI":"10.1086\/595295","volume":"199","author":"PG Kremsner","year":"2009","unstructured":"Kremsner PG, Valim C, Missinou MA, Olola C, Krishna S, Issifou S, Kombila M, Bwanaisa L, Mithwani S, Newton CR, Agbenyega T, Pinder M, Bojang K, Wypij D, Taylor T: Prognostic value of circulating pigmented cells in african children with malaria. J Infect Dis. 2009, 199: 142-150. 10.1086\/595295.","journal-title":"J Infect Dis"},{"key":"1664_CR4","doi-asserted-by":"crossref","first-page":"360","DOI":"10.1093\/ajcp\/86.3.360","volume":"86","author":"C Lawrence","year":"1986","unstructured":"Lawrence C, Olson JA: Birefringent hemozoin identifies malaria. Am J Clin Pathol. 1986, 86: 360-363.","journal-title":"Am J Clin Pathol"},{"key":"1664_CR5","doi-asserted-by":"publisher","first-page":"255","DOI":"10.1186\/1475-2875-8-255","volume":"8","author":"T Hanscheid","year":"2009","unstructured":"Hanscheid T, Frita R, Langin M, Kremsner P, Grobusch M: Is flow cytometry better in counting malaria pigment-containing leukocytes compared to microscopy?. Malar J. 2009, 8: 255-10.1186\/1475-2875-8-255.","journal-title":"Malar J"},{"key":"1664_CR6","doi-asserted-by":"publisher","first-page":"539","DOI":"10.1002\/cyto.990080602","volume":"8","author":"BG de Grooth","year":"1987","unstructured":"de Grooth BG, Terstappen LW, Puppels GJ, Greve J: Light-scattering polarization measurements as a new parameter in flow cytometry. Cytometry. 1987, 8: 539-544. 10.1002\/cyto.990080602.","journal-title":"Cytometry"},{"key":"1664_CR7","doi-asserted-by":"publisher","first-page":"499","DOI":"10.1046\/j.1365-2141.1999.01199.x","volume":"104","author":"BV Mendelow","year":"1999","unstructured":"Mendelow BV, Lyons C, Nhlangothi P, Tana M, Munster M, Wypkema E, Liebowitz L, Marshall L, Scott S, Coetzer TL: Automated malaria detection by depolarization of laser light. Br J Haematol. 1999, 104: 499-503. 10.1046\/j.1365-2141.1999.01199.x.","journal-title":"Br J Haematol"},{"key":"1664_CR8","doi-asserted-by":"publisher","first-page":"1074","DOI":"10.1046\/j.1360-2276.2003.01135.x","volume":"8","author":"I Suh","year":"2003","unstructured":"Suh I, Kim H, Kim J, Lee S, An S, Kim W, Lim C: Evaluation of the Abbott Cell-Dyn 4000 hematology analyzer for detection and therapeutic monitoring of Plasmodium vivax in the Republic of Korea. Trop Med Int Health. 2003, 8: 1074-1081. 10.1046\/j.1360-2276.2003.01135.x.","journal-title":"Trop Med Int Health"},{"key":"1664_CR9","doi-asserted-by":"publisher","first-page":"109","DOI":"10.1186\/1475-2875-7-109","volume":"7","author":"T Hanscheid","year":"2008","unstructured":"Hanscheid T, Langin M, Lell B, Potschke M, Oyakhirome S, Kremsner P, Grobusch M: Full blood count and haemozoin-containing leukocytes in children with malaria: diagnostic value and association with disease severity. Malar J. 2008, 7: 109-10.1186\/1475-2875-7-109.","journal-title":"Malar J"},{"key":"1664_CR10","doi-asserted-by":"publisher","first-page":"225","DOI":"10.1002\/ajh.21690","volume":"85","author":"AH Shankar","year":"2010","unstructured":"Shankar AH, Fawzi WW: Moving toward hematological predictors of disease severity in malaria: going with the flow. Am J Hematol. 2010, 85: 225-226. 10.1002\/ajh.21690.","journal-title":"Am J Hematol"},{"key":"1664_CR11","doi-asserted-by":"publisher","first-page":"125","DOI":"10.1017\/S0031182001008216","volume":"123","author":"E Schwarzer","year":"2001","unstructured":"Schwarzer E, Bellomo G, Giribaldi G, Ulliers D, Arese P: Phagocytosis of malarial pigment hemozoin by human monocytes: a confocal microscopy study. Parasitology. 2001, 123: 125-131. 10.1017\/S0031182001008216.","journal-title":"Parasitology"},{"key":"1664_CR12","doi-asserted-by":"publisher","first-page":"244","DOI":"10.1189\/jlb.0307191","volume":"82","author":"D Strauss-Ayali","year":"2007","unstructured":"Strauss-Ayali D, Conrad SM, Mosser DM: Monocyte subpopulations and their differentiation patterns during infection. J Leukoc Biol. 2007, 82: 244-252. 10.1189\/jlb.0307191.","journal-title":"J Leukoc Biol"},{"key":"1664_CR13","doi-asserted-by":"publisher","first-page":"609","DOI":"10.1016\/j.imbio.2006.05.025","volume":"211","author":"F Tacke","year":"2006","unstructured":"Tacke F, Randolph GJ: Migratory fate and differentiation of blood monocyte subsets. Immunobiology. 2006, 211: 609-618. 10.1016\/j.imbio.2006.05.025.","journal-title":"Immunobiology"},{"key":"1664_CR14","doi-asserted-by":"publisher","first-page":"152","DOI":"10.1111\/j.1365-3083.2007.02051.x","volume":"67","author":"J Skrzeczy\u0144ska-Moncznik","year":"2008","unstructured":"Skrzeczy\u0144ska-Moncznik J, Bzowska M, Loseke S, Grage-Griebenow E, Zembala M, Pryjma J: Peripheral blood CD14high CD16+ monocytes are main producers of IL-10. Scand J Immunol. 2008, 67: 152-159.","journal-title":"Scand J Immunol"},{"key":"1664_CR15","doi-asserted-by":"publisher","first-page":"2173","DOI":"10.1128\/IAI.01078-09","volume":"78","author":"LA Ogonda","year":"2010","unstructured":"Ogonda LA, Orago ASS, Otieno MF, Adhiambo C, Otieno W, Stoute JA: The levels of CD16\/Fc{gamma} receptor IIIA on CD14+ CD16+ monocytes are higher in children with severe Plasmodium falciparum anemia than in children with cerebral or uncomplicated Malaria. Infect Immun. 2010, 78: 2173-2181. 10.1128\/IAI.01078-09.","journal-title":"Infect Immun"},{"key":"1664_CR16","doi-asserted-by":"publisher","first-page":"e1000631","DOI":"10.1371\/journal.ppat.1000631","volume":"5","author":"P Chimma","year":"2009","unstructured":"Chimma P, Roussilhon C, Sratongno P, Ruangveerayuth R, Pattanapanyasat K, P\u00e9rignon JL, Roberts DJ, Druilhe P: A distinct peripheral blood monocyte phenotype is associated with parasite inhibitory activity in acute uncomplicated Plasmodium falciparum malaria. PLoS Pathog. 2009, 5: e1000631-10.1371\/journal.ppat.1000631.","journal-title":"PLoS Pathog"},{"key":"1664_CR17","doi-asserted-by":"publisher","first-page":"1333","DOI":"10.1016\/S0140-6736(82)92403-5","volume":"1","author":"KH Rieckmann","year":"1982","unstructured":"Rieckmann KH: Visual in-vitro test for determining the drug sensitivity of Plasmodium falciparum. Lancet. 1982, 1: 1333-1335. 10.1016\/S0140-6736(82)92403-5.","journal-title":"Lancet"},{"key":"1664_CR18","doi-asserted-by":"publisher","first-page":"4410","DOI":"10.4049\/jimmunol.172.7.4410","volume":"172","author":"C Sunderk\u00f6tter","year":"2004","unstructured":"Sunderk\u00f6tter C, Nikolic T, Dillon MJ, van Rooijen N, Stehling M, Drevets DA, Leenen PJM: Subpopulations of mouse blood monocytes differ in maturation stage and inflammatory response. J Immunol. 2004, 172: 4410-4417.","journal-title":"J Immunol"},{"key":"1664_CR19","unstructured":"Aging study: Peripheral blood leukocytes (PBL profiles) in 32 inbred strains of mice. [http:\/\/phenome.jax.org\/db\/q?rtn=projects\/docstatic&doc=Petkova1\/Petkova1_Protocol]"},{"key":"1664_CR20","doi-asserted-by":"publisher","first-page":"133","DOI":"10.1002\/1097-0320(20011001)45:2<133::AID-CYTO1155>3.0.CO;2-K","volume":"45","author":"B Kramer","year":"2001","unstructured":"Kramer B, Grobusch M, Suttorp N, Neukammer J, Rinneberg H: Relative frequency of malaria pigment-carrying monocytes of nonimmune and semi-immune patients from flow cytometric depolarized side scatter. Cytometry. 2001, 45: 133-140. 10.1002\/1097-0320(20011001)45:2<133::AID-CYTO1155>3.0.CO;2-K.","journal-title":"Cytometry"},{"key":"1664_CR21","unstructured":"Particle discriminator and method, United States Patent 5017497. [http:\/\/www.freepatentsonline.com\/5017497.html]"},{"key":"1664_CR22","unstructured":"How to Calculate Standard Patent Expiry Dates and Data Exclusivity in Key Territories. [http:\/\/www.genericsweb.com\/How_to_Calculate_Standard_Patent_Expiry_Dates_and_Data_Exclusivity.pdf]"},{"key":"1664_CR23","doi-asserted-by":"publisher","first-page":"4912","DOI":"10.1128\/IAI.00455-10","volume":"78","author":"G Giribaldi","year":"2010","unstructured":"Giribaldi G, Prato M, Ulliers D, Gallo V, Schwarzer E, Akide-Ndunge OB, Valente E, Saviozzi S, Calogero RA, Arese P: Involvement of inflammatory chemokines in survival of human monocytes fed with malarial pigment. Infect Immun. 2010, 78: 4912-4921. 10.1128\/IAI.00455-10.","journal-title":"Infect Immun"},{"key":"1664_CR24","doi-asserted-by":"publisher","first-page":"2859","DOI":"10.1182\/blood.V98.9.2859","volume":"98","author":"BC Urban","year":"2001","unstructured":"Urban BC, Mwangi T, Ross A, Kinyanjui S, Mosobo M, Kai O, Lowe B, Marsh K, Roberts DJ: Peripheral blood dendritic cells in children with acute Plasmodium falciparum malaria. Blood. 2001, 98: 2859-2861. 10.1182\/blood.V98.9.2859.","journal-title":"Blood"},{"key":"1664_CR25","doi-asserted-by":"crossref","first-page":"1601","DOI":"10.1128\/IAI.66.4.1601-1606.1998","volume":"66","author":"E Schwarzer","year":"1998","unstructured":"Schwarzer E, Alessio M, Ulliers D, Arese P: Phagocytosis of the malarial pigment, hemozoin, impairs expression of major histocompatibility complex class II antigen, CD54, and CD11c in human monocytes. Infect Immun. 1998, 66: 1601-1606.","journal-title":"Infect Immun"},{"key":"1664_CR26","doi-asserted-by":"publisher","first-page":"e10","DOI":"10.1182\/blood-2009-07-235028","volume":"115","author":"MA Ingersoll","year":"2010","unstructured":"Ingersoll MA, Spanbroek R, Lottaz C, Gautier EL, Frankenberger M, Hoffmann R, Lang R, Haniffa M, Collin M, Tacke F, Habenicht AJ, Ziegler-Heitbrock L, Randolph GJ: Comparison of gene expression profiles between human and mouse monocyte subsets. Blood. 2010, 115: e10-19. 10.1182\/blood-2009-07-235028.","journal-title":"Blood"},{"key":"1664_CR27","doi-asserted-by":"publisher","first-page":"227","DOI":"10.1002\/ajh.21653","volume":"85","author":"GC Davenport","year":"2010","unstructured":"Davenport GC, Ouma C, Hittner JB, Were T, Ouma Y, Ong'echa JM, Perkins DJ: Hematological predictors of increased severe anemia in Kenyan children coinfected with Plasmodium falciparum and HIV-1. Am J Hematol. 2010, 85: 227-233. 10.1002\/ajh.21653.","journal-title":"Am J Hematol"},{"key":"1664_CR28","doi-asserted-by":"publisher","first-page":"e8446","DOI":"10.1371\/journal.pone.0008446","volume":"4","author":"AA Lamikanra","year":"2009","unstructured":"Lamikanra AA, Theron M, Kooij TWA, Roberts DJ: Hemozoin (malarial pigment) directly promotes apoptosis of erythroid precursors. PLoS ONE. 2009, 4: e8446-10.1371\/journal.pone.0008446.","journal-title":"PLoS ONE"},{"key":"1664_CR29","volume-title":"Field application of in vitro assays for the sensitivity of human malaria parasites to antimalarial drugs","author":"L Basco","year":"2007","unstructured":"Basco L: Field application of in vitro assays for the sensitivity of human malaria parasites to antimalarial drugs. 2007, Geneva: World Health Organization"},{"key":"1664_CR30","doi-asserted-by":"publisher","first-page":"175","DOI":"10.1016\/S1471-4922(03)00028-X","volume":"19","author":"H Noedl","year":"2003","unstructured":"Noedl H, Wongsrichanalai C, Wernsdorfer WH: Malaria drug-sensitivity testing: new assays, new perspectives. Trends Parasitol. 2003, 19: 175-181. 10.1016\/S1471-4922(03)00028-X.","journal-title":"Trends Parasitol"},{"key":"1664_CR31","doi-asserted-by":"publisher","first-page":"1651","DOI":"10.1128\/JCM.02250-09","volume":"48","author":"S Wein","year":"2010","unstructured":"Wein S, Maynadier M, Tran Van Ba C, Cerdan R, Peyrottes S, Fraisse L, Vial H: Reliability of antimalarial sensitivity tests depends on drug mechanisms of action. J Clin Microbiol. 2010, 48: 1651-1660. 10.1128\/JCM.02250-09.","journal-title":"J Clin Microbiol"},{"key":"1664_CR32","doi-asserted-by":"publisher","first-page":"5452","DOI":"10.1016\/j.bmcl.2009.07.095","volume":"19","author":"BT Grimberg","year":"2009","unstructured":"Grimberg BT, Jaworska MM, Hough LB, Zimmerman PA, Phillips JG: Addressing the malaria drug resistance challenge using flow cytometry to discover new antimalarials. Bioorg Med Chem Lett. 2009, 19: 5452-5457. 10.1016\/j.bmcl.2009.07.095.","journal-title":"Bioorg Med Chem Lett"},{"key":"1664_CR33","doi-asserted-by":"publisher","first-page":"144","DOI":"10.1016\/j.exppara.2008.10.008","volume":"121","author":"S Izumiyama","year":"2009","unstructured":"Izumiyama S, Omura M, Takasaki T, Ohmae H, Asahi H: Plasmodium falciparum: Development and validation of a measure of intraerythrocytic growth using SYBR Green I in a flow cytometer. Exp Parasitol. 2009, 121: 144-150. 10.1016\/j.exppara.2008.10.008.","journal-title":"Exp Parasitol"},{"key":"1664_CR34","doi-asserted-by":"publisher","first-page":"138","DOI":"10.1016\/0169-4758(95)80133-2","volume":"11","author":"CJ Janse","year":"1995","unstructured":"Janse CJ, Waters AP: Plasmodium berghei: The application of cultivation and purification techniques to molecular studies of malaria parasites. Parasitol Today. 1995, 11: 138-143. 10.1016\/0169-4758(95)80133-2.","journal-title":"Parasitol Today"}],"container-title":["Malaria Journal"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/1475-2875-10-74.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,9,1]],"date-time":"2021-09-01T13:41:08Z","timestamp":1630503668000},"score":1,"resource":{"primary":{"URL":"https:\/\/malariajournal.biomedcentral.com\/articles\/10.1186\/1475-2875-10-74"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2011,3,31]]},"references-count":34,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2011,12]]}},"alternative-id":["1664"],"URL":"https:\/\/doi.org\/10.1186\/1475-2875-10-74","relation":{},"ISSN":["1475-2875"],"issn-type":[{"value":"1475-2875","type":"electronic"}],"subject":[],"published":{"date-parts":[[2011,3,31]]},"assertion":[{"value":"1 February 2011","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"31 March 2011","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"31 March 2011","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}}],"article-number":"74"}}