Flow Cytometry-Based Methods for Measurement of Cytosolic Calcium and Surface Protein Expression in Plasmodium falciparum Merozoites

Author(s):  
Shailja Singh ◽  
Chetan E. Chitnis
2022 ◽  
Author(s):  
Viola Introini ◽  
Alejandro Marin-Menendez ◽  
Guilherme Nettesheim ◽  
Yen-Chun Lin ◽  
Silvia N Kariuki ◽  
...  

Malaria parasites such as Plasmodium falciparum have exerted formidable selective pressures on the human genome. Of the human genetic variants associated with malaria protection, beta thalassaemia (a haemoglobinopathy) was the earliest to be associated with malaria prevalence. However, the malaria protective properties of beta thalassaemic erythrocytes remain unclear. Here we studied the mechanics and surface protein expression of beta thalassaemia heterozygous erythrocytes, measured their susceptibility to P. falciparum invasion, and calculated the energy required for merozoites to invade them. We found invasion-relevant differences in beta thalassaemic cells versus matched controls, specifically: elevated membrane tension, reduced bending modulus, and higher levels of expression of the major invasion receptor basigin. However, these differences acted in opposition to each other with respect to their likely impact on invasion, and overall we did not observe beta thalassaemic cells to have lower P. falciparum invasion efficiency for any of the strains tested.


Lab on a Chip ◽  
2017 ◽  
Vol 17 (7) ◽  
pp. 1349-1358 ◽  
Author(s):  
Rhonda Jack ◽  
Khadijah Hussain ◽  
Danika Rodrigues ◽  
Mina Zeinali ◽  
Ebrahim Azizi ◽  
...  

In light of the significance of tumor cell heterogeneity, we describe a facile workflow to isolate distinct groups of tumor cells immunomagnetically, according to their surface-protein expression levels.


Luminescence ◽  
2007 ◽  
Vol 22 (3) ◽  
pp. 163-170 ◽  
Author(s):  
Roope J. Huttunen ◽  
Tomás C. O'Riordan ◽  
Pirkko L. Härkönen ◽  
Juhani T. Soini ◽  
Niko J. Meltola ◽  
...  

2006 ◽  
Vol 52 (6) ◽  
pp. 1080-1088 ◽  
Author(s):  
Ethan Fitzpatrick ◽  
Sterling McBride ◽  
Jonathan Yavelow ◽  
Saltanat Najmi ◽  
Peter Zanzucchi ◽  
...  

Abstract Background: The analysis of single cells obtained from needle aspirates of tumors is constrained by the need for processing. To this end, we investigated two microfluidic approaches to measure the expression of surface proteins in single cancer cells or in small populations (<50 cells). Methods: One approach involved indirect fluorescence labeling of cell-surface proteins and channeling of cells in a microfluidic device past a fluorescence detector for signal quantification and analysis. A second approach channeled cells in a microfluidic device over detection zones coated with ligands to surface proteins and measured rates of passage and of retardation based on transient interactions between surface proteins and ligands. Results: The fluorescence device detected expression of integrin α5 induced by basic fibroblast growth factor (FGF-2) treatment in MCF-7 cells and that of Her-2/neu in SK-BR-3 cells compared with controls. Experiments measuring passage retardation showed significant differences in passage rates between FGF-2–treated and untreated MCF-7 cells over reaction regions coated with fibronectin and antibody to integrin α5β1 compared with control regions. Blocking peptides reversed the retardation, demonstrating specificity. Conclusions: Immunofluorescence detection in a microfluidic channel demonstrates the potential for assaying surface protein expression in a few individual cells and will permit the development of future iterations not requiring cell handling. The flow retardation device represents the first application of this technology for assessing cell-surface protein expression in cancer cells and may provide a way for analyzing expression profiles of single cells without preanalytical manipulation.


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