Background: Microscopic analysis of blood smears is currently the most frequently used method to measure parasitemias in experiments of drug efficacy in murine models of malaria. However, it is subjective and labour intensive, which preclude its utilization in large-scale evaluation programs. Flow cytometry is an alternative method, but due to the limited specificity achieved with the currently available techniques, it has not been widely used in murine models of malaria during preclinical evaluation. We describe a new flow cytometric method based on the differences of autofluorescence and DNA content measured after staining with YOYO-1 that are observed in infected erythrocytes compared with noninfected erythrocytes. Methods: Samples of blood from Plasmodium yoeliiinfected animals were fixed with glutaraldehyde, incubated with RNAase, and stained with YOYO-1 in 96-well plate format. After acquisition, erythrocytes gated in loga-

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Many articles dealing with individual cell lag phase determination assume that growth, when observed, comes from one cell. This assumption is not in agreement with the Poisson distribution, which uses the probability of growth in a sample to predict how many samples contain one, two, or some other number of cells. This article analyses and compares different approaches to improve the accuracy of lag phase estimation of individual cells and micropopulations. It argues that if the highest initial load, as predicted by the Poisson distribution, is assigned to the sample with the shortest lag phase, the second highest to the sample with the second shortest lag phase and so on, the resulting lag phase distributions would be more accurate. This study also proposes the use of a robust test, permutation test, to compare lag phase distributions obtained in different situations.

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