{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T20:04:48Z","timestamp":1792008288656,"version":"4.4.0"},"reference-count":33,"publisher":"Cambridge University Press (CUP)","issue":"1","license":[{"start":{"date-parts":[[2021,9,7]],"date-time":"2021-09-07T00:00:00Z","timestamp":1630972800000},"content-version":"unspecified","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["cambridge.org"],"crossmark-restriction":true},"short-container-title":["Parasitology"],"published-print":{"date-parts":[[2022,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Flow cytometry analysis emerges as an alternative methodology to microscopy for determination of the\n                    <jats:italic>Leishmania<\/jats:italic>\n                    -infection rates of macrophages. Various flow cytometric approaches have been established for the quantification of\n                    <jats:italic>Leishmania<\/jats:italic>\n                    parasites within host cells, labelled either directly fluorescent dyes or indirectly with fluorescently conjugated antibodies. Although these techniques allow accurate quantification of infection, they fail at detection of non-infected macrophages specifically. This study introduces a new flow cytometric approach for the determination of infection rates of macrophages infected by\n                    <jats:italic>Leishmania infantum<\/jats:italic>\n                    parasites. Prior to infection, J774A.1 macrophages and\n                    <jats:italic>L. infantum<\/jats:italic>\n                    promastigotes were stained separately with PKH26 and PKH67 dyes, respectively. Dual staining enabled detection of each cell type, where non-infected macrophages were also recorded for the quantification. Dual-PKH staining achieved high success in selective staining of promastigotes (99.71%) and macrophages (99.57%). The percentages of parasite-infected macrophages were determined for initial 1:2.5 and 1:10 infection ratios as 15.68 and 61.70%, respectively; indicating significant increase in infection rate parallel to the initial treatment ratio. These results demonstrated that the introduced dual-fluorescence flow cytometric approach can be successfully used as an accurate and rapid quantification method for\n                    <jats:italic>L. infantum<\/jats:italic>\n                    -infected macrophages and strengthens the hypothesis that flow cytometric approaches could replace conventional microscopic methodologies.\n                  <\/jats:p>","DOI":"10.1017\/s0031182021001530","type":"journal-article","created":{"date-parts":[[2021,9,7]],"date-time":"2021-09-07T02:57:46Z","timestamp":1630983466000},"page":"44-50","update-policy":"https:\/\/doi.org\/10.1017\/policypage","source":"Crossref","is-referenced-by-count":7,"title":["Novel dual-fluorescent flow cytometric approach for quantification of macrophages infected with\n                    <i>Leishmania infantum<\/i>\n                    parasites"],"prefix":"10.1017","volume":"149","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4789-9095","authenticated-orcid":false,"given":"Zeynep","family":"Islek","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9434-3861","authenticated-orcid":false,"given":"Mehmet 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\u00a9 The Author(s), 2021. Published by Cambridge University Press","name":"copyright","label":"Copyright","group":{"name":"copyright_and_licensing","label":"Copyright and Licensing"}},{"value":"This is an Open Access article, distributed under the terms of the Creative Commons Attribution licence (http:\/\/creativecommons.org\/licenses\/by\/4.0\/), which permits unrestricted re-use, distribution and reproduction, provided the original article is properly cited.","name":"license","label":"License","group":{"name":"copyright_and_licensing","label":"Copyright and Licensing"}},{"value":"This content has been made available to all.","name":"free","label":"Free to read"}]}}