The automated counting programs are an accurate and practical way of quantifying static parasite binding assays to purified proteins. They are less accurate when applied to cell based systems, but can still provide a reasonable level of accuracy to give a semi-quantitative readout

Barrett, Steve D.

Craig, Alister

Faragher, Brian

Mustaffa, Khairul

Paton, Douglas

Szestak, Tadge

Malaria Journal

PubMed

The automated counting programs are an accurate and practical way of quantifying static parasite binding assays to purified proteins. They are less accurate when applied to cell based systems, but can still provide a reasonable level of accuracy to give a semi-quantitative readout.</p

Mustaffa, Khairul M.F.

LSTM Research Portal

English

Automated counting for Plasmodium falciparum cytoadherence experiments.

Abstract
          
            Background
            The ability of mature forms of Plasmodium falciparum infected erythrocytes to bind to a range of host receptors including those displayed on endothelial cells has been associated with the pathology of this infection. Investigations into this adhesive phenomenon have used protein and cell-based adhesion assays to quantify the ability of infected red blood cells to bind. These adhesion assays tend to have relatively high inherent variability and so require multiple experiments in order to provide good quantitation. This means that investigators doing these experiments must count many fields of adherent parasites, a task that is time-consuming and laborious. To address this issue and to facilitate cytoadherence research, developed automated protocols were developed for counting parasite adhesion.
          
          
            Methods
            Parasite adhesion assays were mainly carried out under static conditions using purified receptors, which is the simplest form of these assays and is translatable to the field. Two different software platforms were used, one commercial (Image Pro-Plus (Media Cybernetics)) and one available in the public domain (ImageSXM) based on the freely available NIH Image software. The adhesion assays were performed and parasite binding quantified using standard manual techniques. Images were also captured using video microscopy and analysed using the two automated systems. The results generated by each system were compared using the Bland and Altman method for assessing the agreement between two methods.
          
          
            Results
            Both automated counting programs showed concordance compared to the 'gold standard' manual counting within the normal range of adhesion seen with these assays, although the ImageSXM technique had some systematic bias. There was some fall-off in accuracy at very high parasite densities, but this can be resolved through good design of the experiments. Cell based assays were also used as inputs to one of the automated systems (ImageSXM) and produced variable, but encouraging, results.
          
          
            Conclusions
            The automated counting programs are an accurate and practical way of quantifying static parasite binding assays to purified proteins. They are less accurate when applied to cell based systems, but can still provide a reasonable level of accuracy to give a semi-quantitative readout.
          </jats:sec

Douglas Paton

Brian Faragher

Khairul MF Mustaffa

Tadge Szestak

Steve D Barrett

Alister G Craig

Crossref

Automated counting for Plasmodium falciparum cytoadherence experiments

Springer - Publisher Connector

BackgroundThe ability of mature forms of Plasmodium falciparum infected erythrocytes to bind to a range of host receptors including those displayed on endothelial cells has been associated with the pathology of this infection. Investigations into this adhesive phenomenon have used protein and cell-based adhesion assays to quantify the ability of infected red blood cells to bind. These adhesion assays tend to have relatively high inherent variability and so require multiple experiments in order to provide good quantitation. This means that investigators doing these experiments must count many fields of adherent parasites, a task that is time-consuming and laborious. To address this issue and to facilitate cytoadherence research, developed automated protocols were developed for counting parasite adhesion.MethodsParasite adhesion assays were mainly carried out under static conditions using purified receptors, which is the simplest form of these assays and is translatable to the field. Two different software platforms were used, one commercial (Image Pro-Plus (Media Cybernetics)) and one available in the public domain (ImageSXM) based on the freely available NIH Image software. The adhesion assays were performed and parasite binding quantified using standard manual techniques. Images were also captured using video microscopy and analysed using the two automated systems. The results generated by each system were compared using the Bland and Altman method for assessing the agreement between two methods.ResultsBoth automated counting programs showed concordance compared to the 'gold standard' manual counting within the normal range of adhesion seen with these assays, although the ImageSXM technique had some systematic bias. There was some fall-off in accuracy at very high parasite densities, but this can be resolved through good design of the experiments. Cell based assays were also used as inputs to one of the automated systems (ImageSXM) and produced variable, but encouraging, results.ConclusionsThe automated counting programs are an accurate and practical way of quantifying static parasite binding assays to purified proteins. They are less accurate when applied to cell based systems, but can still provide a reasonable level of accuracy to give a semi-quantitative readout

Mustaffa, Khairul MF

Barrett, Steve D

Craig, Alister G

University of Liverpool Repository

Automated counting for <i>Plasmodium falciparum</i> cytoadherence experiments

LSTM Online Archive

Adhesion of Plasmodium falciparum-infected erythrocytes to human cells: molecular mechanisms and therapeutic implications. Expert Rev Mol Med

Characterization of Plasmodium falciparum-infected erythrocyte and Pselectin interaction under flow conditions. Blood

Promiscuity of clinical Plasmodium falciparum isolates for multiple adhesion molecules under flow conditions.

Receptor-specific adhesion and clinical disease in Plasmodium falciparum. Am J Trop Med Hyg

Snow RW: Estimating the global clinical burden of Plasmodium falciparum malaria in 2007. PLoS Med

http://archive.lstmed.ac.uk/2016/1/Craig-Malaria-J-11.pdf

Automated counting for Plasmodium falciparum cytoadherence experiments.

Abstract

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LSTM Research Portal

Crossref

Springer - Publisher Connector

LSTM Research Portal

University of Liverpool Repository

Springer - Publisher Connector

LSTM Online Archive