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Flow cytometric assessment of Leishmania spp metacyclic differentiation: validation by morphological features and specific markers.
- Source :
-
Experimental parasitology [Exp Parasitol] 2005 May; Vol. 110 (1), pp. 39-47. - Publication Year :
- 2005
-
Abstract
- Characterization of infective metacyclic promastigotes of Leishmania spp can be an essential step in several experimental protocols. Metacyclic forms of all Leishmania species display a typical morphology with short, narrow cell body, and an elongated flagellum. This feature suggests that metacyclics can be distinguished from procyclic forms by non-fluorimetric flow cytometric parameters thus enabling the follow-up of their appearance and acquisition of specific properties, during metacyclogenesis in in vitro cultures. Here we describe the flow cytometric parameters of stage-specific promastigotes of Leishmania major, Leishmania donovani, Leishmania amazonensis, and Leishmania braziliensis. Our findings were validated by optical microscopy morphology and specific procyclic labeling with FITC-peanut agglutinin. Furthermore, we show that parasite's distribution in the plot during differentiation in culture is not species specific and that the parasites displaying low forward-angle light scatter (FSC(low)) are three times more infective than the FSC(high) ones. The method here described can be applied to the identification of metacyclics of different Leishmania spp within the whole stationary population.
- Subjects :
- Animals
Cells, Cultured
Flow Cytometry
Humans
Leishmania isolation & purification
Leishmania ultrastructure
Leishmania braziliensis growth & development
Leishmania braziliensis isolation & purification
Leishmania braziliensis ultrastructure
Leishmania donovani growth & development
Leishmania donovani isolation & purification
Leishmania donovani ultrastructure
Leishmania major growth & development
Leishmania major isolation & purification
Leishmania major ultrastructure
Leishmania mexicana growth & development
Leishmania mexicana isolation & purification
Leishmania mexicana ultrastructure
Life Cycle Stages
Macrophages, Peritoneal parasitology
Mice
Mice, Inbred BALB C
Reproducibility of Results
Species Specificity
Leishmania growth & development
Subjects
Details
- Language :
- English
- ISSN :
- 0014-4894
- Volume :
- 110
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Experimental parasitology
- Publication Type :
- Academic Journal
- Accession number :
- 15804377
- Full Text :
- https://doi.org/10.1016/j.exppara.2005.01.004