Part I. Collecting, growing, monitoring, and manipulating Plasmodium falciparum parasites 1. Identification and enumeration of Plasmodium falciparum parasites by light microscopy Augustina Frimpong, Frederica D Partey and Michael F Ofori 2. Collection and cryopreservation of Plasmodium falciparum clinical isolates in the field Frederica D Partey, Augustina Frimpong and Michael F Ofori 3. Preservation of parasite RNA in the field Bernard Tornyigah and Nicaise Tuikue Ndam 4. Preservation and extraction of malaria parasite DNA from dried blood spots Helle Hansson, Queen Saidi and Michael Alifrangis 5. Establishing and maintaining in vitro cultures of asexual blood stages of Plasmodium falciparum Mary Lopez-Perez and Zakaria Seidu 6. Genotyping of Plasmodium falciparum isolates to assess clone composition in parasite cultures Ioanna Broumou, David F Plaza and Anna Färnert 7. Immunomagnetic selection of Plasmodium falciparum-infected erythrocytes expressing particular PfEMP1 variants Mary Lopez-Perez and Rebecca W Olsen 8. Single-cell sorting of Plasmodium falciparum-infected erythrocytes expressing particular PfEMP1 variants Mary Lopez-Perez 9. Selecting Plasmodium falciparum infected erythrocytes for adhesion to cell lines Yvonne Azasi and J Alexandra Rowe 10. Synchronisation of Plasmodium falciparum and P. knowlesi in vitro cultures using a highly specific protein kinase inhibitor Margarida Ressurreição, Robert W Moon, David A Baker, Christiaan van Ooij 11. Generation of Plasmodium falciparum gametocytes in vitro with specific considerations for field isolates Bismarck Dinko, Richmond Tackie, Gisela Henriques, Lindsay Stewart and Colin Sutherland 12. Imaging of extracellular vesicles derived from Plasmodium falciparum-infected red blood cells using atomic force microscopy Irit Rosenhek-Goldian, Paula Abou Karam, Neta Regev-Rudzki and Alicia Rojas Part II. Plasmodium falciparum gene transcription and manipulation of parasite genomes 13. Analysis of var gene transcript patterns by quantitative real-time PCR Anna Bachmann and Thomas Lavstsen 14. Analysis of var gene transcription pattern using DBLa-tags Kivisi C Andisi and Abdirahman I Abdi 15. RNAseq of infected erythrocyte surface antigen-encoding genes Hanh Hong Thi Nguyen, Suffian Azizan, Lee Ming Yeoh, Jingyi Tang and Michael F Duffy 16. Resetting var gene transcription in Plasmodium falciparum Vera Mitesser and Ron Dzikowski 17. CRISPR/Cas9 editing of the Plasmodium falciparum genome Sophie Adjalley and Marcus CS Lee 18. CRISPR-Cas9 editing of the Plasmodium falciparum genome - Special applications Xu Zhang, Kirk W Deitsch and Ron Dzikowski Part III. Plasmodium falciparum-infected erythrocyte surface antigens 19. Extraction and immunoprecipitation of VAR2CSA, the PfEMP1 associated with placental malaria Benoit Gamain and Dominique Dorin-Semblat 20. Expression of single-domain soluble and disulfide-folded PfEMP1 antigens in the Escherichia coli SHuffle expression system Maria del Pilar Quintana 21. Expression of large full-length PfEMP1 proteins in HEK293 cells Jonathan P Renn, Justin YA Doritchamou and Patrick E Duffy 22. Receptor affinity-based purification of PfEMP1 proteins Rebecca W Olsen, Jennifer Suurbaar and Anja R Jensen 23. Analysis of antibody reactivity to malaria antigens by microsphere-based multiplex immunoassay Isobel S Walker, Amy WT Chung, T Damelang and Stephen J Rogerson 24. High-throughput BioPlex assay for the study of functionally active Plasmodium falciparum antigens that are expressed on the surface of infected erythrocytes Andrew V. Oleinikov 25. Protein microarrays as a tool to analyze antibody responses to variant surface antigens expressed on the surface of Plasmodium falciparum-infected erythrocytes Albert E Zhou, Aarti Jain, Rie Nakajima, Biraj Shrestha, Emily M Stucke, Sudhaunshu Joshi, Kathy A Strauss, Per N Hedde, Andrea A-Y Berry, PL Felgner and Mark A Travassos 26. Protocol for differential biopanning of Plasmodium falciparum phage display cDNA library to identify parasite targets of protective antibodies Jenna Zuromski, Jonathan Kurtis and Dipak Kumar Raj Part IV. Antibodies to Plasmodium falciparum-infected erythrocyte surface antigens 27. Affinity purification of PfEMP1-specific antibodies from human blood Rebecca W Olsen, Jennifer Suurbaar and Anja R Jensen 28. Production of anti-PfEMP1 polyclonal antisera in rats and mice Rebecca W Olsen, Jennifer Suurbaar and Anja R Jensen 29. Production of PfEMP1-specific mouse monoclonal antibodie Nanna Dalgaard and Lea Barfod 30. Production of PfEMP1-specific human monoclonal antibodies from naturally immune individuals Melanie Walker and Lea Barfod Part V. Analysis of antigens on Plasmodium falciparum-infected erythrocytes 31. Visualization of infected red blood cell surface antigens by fluorescence microscop Markus Ganter, Julien Guizetti and Nicole Kilian 32. Analysis by flow cytometry of a2-macroglobulin and non-immune IgM-binding to Plasmodium falciparum-infected erythrocytes Lars Hviid and Mary Lopez-Perez 33. An improved method for assessing antigen presentation on the surface of Plasmodium falciparum-infected erythrocytes by immuno-electron microscopy Marek Cyrklaff, Cecilia P Sanchez, Lukas Hanebutte, Julia Jäger, Ulrich S. Schwartz and Michael Lanzer 34. Assessing antigen presentation on the surface of Plasmodium falciparum-infected erythrocytes by photoactivated localization microscopy (PALM) Christos Karathanasis, Cecilia P Sanchez, Mike Heilemann and Michael Lanzer 35. Surface plasmon resonance analysis of PfEMP1 interaction with receptors Frank Lennartz and Matthew K. Higgins Part VI. Analysis of P. falciparum-infected erythrocyte adhesion 36. Analysis of antibody neutralization of PfEMP1 binding by competition ELISA Rebecca W Olsen, Jennifer Suurbaar and Anja R Jensen 37. Measuring rosetting inhibition in Plasmodium falciparum parasites using a flow cytometry-based assay Maria del Pilar Quintana and Jun-Hong Ch'ng 38. Bridging and clumping: investigating platelet interactions with Plasmodium falciparum-infected red blood cells and endothelial cells in cerebral malaria Samuel C Wassmer, George E Grau and Ian J MacCormick 39. Assay of static adhesion of Plasmodium falciparum-infected erythrocytes to cells, including inhibition of the adhesion Yvonne Azasi 40. Static adhesion of Plasmodium falciparum-infected erythrocytes to purified and recombinant receptors Janet Storm and Alister G Craig 41. Selecting Plasmodium falciparum-infected erythrocytes for adhesion to recombinant receptors under flow conditions Yvonne Adams 42. Chip-based assay of adhesion of Plasmodium falciparum-infected erythrocytes to cells under flow Yvonne Adams and Anja R Jensen 43. Binding of Plasmodium falciparum-infected red blood cells to engineered 3D microvessels Livia Piatti, Caitlin Howard, Ying Zheng and Maria Bernabeu 44. 3D organoid assay of the impact of Plasmodium falciparum-infected erythrocyte adhesion on the blood-brain barrier Yvonne Adams and Anja R Jensen 45. Receptor-functionalized lipid membranes as biomimetic surfaces for adhesion of Plasmodium falciparum-infected erythrocytes Motom
