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. 2019 Sep 17;9:13436. doi: 10.1038/s41598-019-49895-3

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Schematic of experimental setup and SMOOT_{human skin} analysis. (A) Schematic of an Anopheles mosquito as the host of Plasmodium sporozoites within its salivary glands. Isolated sporozoites, Pf ^WT or Pf ^RA, were injected into human skin (1). The skin samples containing sporozoites were filmed using a confocal microscope (2) (Images of needle and microscope were adapted from image copyright https://smart.servier.com, Creative Commons Attribution 3.0 Unported License, https://creativecommons.org/licenses/by/3.0/). (B) Raw confocal video images were uploaded into SMOOT_{human skin}. Per video frame individual sporozoites were semi-automatically segmented (1). Segmented sporozoites in consecutive frames were stitched to generate tracks. Generated tracks have a unique sporozoite ID (i) in order to extract measured parameters (for example movement pattern (ii)) per sporozoite over time (2). (C) Sporozoite tracks are divided into segments based on the underlying movement pattern.