title: In vivo photoacoustic flow cytometry for early malaria diagnosis.
abstract: In vivo photoacoustic (PA) flow cytometry (PAFC) has already demonstrated a great potential for the diagnosis of deadly diseases through ultrasensitive detection of rare disease-associated circulating markers in whole blood volume. Here, we demonstrate the first application of this powerful technique for early diagnosis of malaria through label-free detection of malaria parasite-produced hemozoin in infected red blood cells (iRBCs) as high-contrast PA agent. The existing malaria tests using blood smears can detect the disease at 0.001-0.1% of parasitemia. On the contrary, linear PAFC showed a potential for noninvasive malaria diagnosis at an extremely low level of parasitemia of 0.0000001%, which is ∼10(3) times better than the existing tests. Multicolor time-of-flight PAFC with high-pulse repetition rate lasers at wavelengths of 532, 671, and 820 nm demonstrated rapid spectral and spatial identification and quantitative enumeration of individual iRBCs. Integration of PAFC with fluorescence flow cytometry (FFC) provided real-time simultaneous detection of single iRBCs and parasites expressing green fluorescence proteins, respectively. A combination of linear and nonlinear nanobubble-based multicolor PAFC showed capability to real-time control therapy efficiency by counting of iRBCs before, during, and after treatment. Our results suggest that high-sensitivity, high-resolution ultrafast PAFC-FFC platform represents a powerful research tool to provide the insight on malaria progression through dynamic study of parasite-cell interactions directly in bloodstream, whereas portable hand-worn PAFC device could be broadly used in humans for early malaria diagnosis. © 2016 International Society for Advancement of Cytometry.
authors.affiliation: Arkansas Nanomedicine Center, University of Arkansas for Medical Sciences, Little Rock, Arkansas, 72205.|Division of Biochemical Toxicology, National Center for Toxicological Research, U.S. Food and Drug Administration, Jefferson, Arkansas, 72079.
authors.forename: Chengzhong
authors.initials: C
authors.lastname: Cai
authors.affiliation: Arkansas Nanomedicine Center, University of Arkansas for Medical Sciences, Little Rock, Arkansas, 72205.
authors.forename: Kai A
authors.initials: KA
authors.lastname: Carey
authors.affiliation: Arkansas Nanomedicine Center, University of Arkansas for Medical Sciences, Little Rock, Arkansas, 72205.
authors.forename: Dmitry A
authors.initials: DA
authors.lastname: Nedosekin
authors.affiliation: Arkansas Nanomedicine Center, University of Arkansas for Medical Sciences, Little Rock, Arkansas, 72205.
authors.forename: Yulian A
authors.initials: YA
authors.lastname: Menyaev
authors.affiliation: Arkansas Nanomedicine Center, University of Arkansas for Medical Sciences, Little Rock, Arkansas, 72205.
authors.forename: Mustafa
authors.initials: M
authors.lastname: Sarimollaoglu
authors.affiliation: Arkansas Nanomedicine Center, University of Arkansas for Medical Sciences, Little Rock, Arkansas, 72205.
authors.forename: Ekaterina I
authors.initials: EI
authors.lastname: Galanzha
authors.affiliation: Department of Microbiology and Immunology, University of Arkansas for Medical Sciences, Little Rock, Arkansas, 72205.
authors.forename: Jason S
authors.initials: JS
authors.lastname: Stumhofer
authors.affiliation: Arkansas Nanomedicine Center, University of Arkansas for Medical Sciences, Little Rock, Arkansas, 72205.
authors.forename: Vladimir P
authors.initials: VP
authors.lastname: Zharov
mesh_terms: D000818:Animals; D034201:Computers, Handheld; D004423:Ear / Q000098:blood supply / Q000469:parasitology; D042241:Early Diagnosis; D004912:Erythrocytes / Q000469:parasitology*; D005434:Flow Cytometry / Q000295:instrumentation / Q000379:methods*; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006420:Hemeproteins / Q000032:analysis* / Q000096:biosynthesis / Q000737:chemistry; D006790:Host-Parasite Interactions; D007834:Lasers; D008288:Malaria / Q000175:diagnosis* / Q000469:parasitology; D051379:Mice; D008810:Mice, Inbred C57BL; D018512:Parasitemia / Q000175:diagnosis* / Q000469:parasitology; D061088:Photoacoustic Techniques / Q000295:instrumentation* / Q000379:methods; D010967:Plasmodium yoelii / Q000254:growth & development* / Q000472:pathogenicity; D052939:Schizonts / Q000737:chemistry / Q000502:physiology
chemical_list: D006420:Hemeproteins; D049452:Green Fluorescent Proteins; C020753:hemozoin
keywords: early diagnosis; fluorescence; hemozoin; in vivo flow cytometry; label-free detection; malaria; nanobubbles; photoacoustic spectroscopy