Please use this identifier to cite or link to this item:
|Title:||Re-evaluating acridine orange for rapid flow cytometric enumeration of parasitemia in malaria-infected rodents|
|Authors:||Sebastian Chakrit Bhakdi|
Hartmut P H Neumann
Universitat Freiburg im Breisgau
Institut Pasteur, Paris
Faculty of Medicine, Siriraj Hospital, Mahidol University
|Keywords:||Biochemistry, Genetics and Molecular Biology;Medicine|
|Citation:||Cytometry Part A. Vol.71, No.9 (2007), 662-667|
|Abstract:||Methods facilitating research in malaria are of pivotal relevance. Flow cytometry offers the possibility of rapid enumeration of parasitemia. It relies on staining the parasite DNA to distinguish between infected and non-infected red blood cell (RBC) populations. Unfortunately, in rodents abundant reticulocyte RNA interferes with the application of the method. This results in time-consuming sample preparation protocols that offer no clear advantage over microscopic counting. We re-evaluated the use of the DNA/RNA discriminating vital fluorochrome acridine orange (AO) for rapid flow cytometric enumeration of parasitemia in rodents. Whole blood from rodents infected with Plasmodium berghei and Plasmodium yoelii was stained with AO and analyzed by flow cytometer. A newly developed two-channel (FL1/FL3) detection method was compared with conventional one-channel (FL1) detection and microscopic counting. The new AO two-channel detection method clearly discriminated between infected and non-infected RBC populations. It showed to be linear above parasitemias of 0.3%. Sample processing time amounted to approximately 5 min. It is shown that AO can be used for rapid, precise, and accurate enumeration of parasitemia in rodents. Due to its ease of handling the method might find widespread application in malaria research. © 2007 International Society for Analytical Cytology.|
|Appears in Collections:||Scopus 2006-2010|
Files in This Item:
There are no files associated with this item.
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.