1. Development of an automated image analysis protocol for quantification of intracellular forms of Leishmania spp
- Author
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Helena Castro, Ana Georgina Gomes-Alves, Ana M. Tomás, Tânia Cruz, André F. Maia, Universidade do Minho, and Instituto de Investigação e Inovação em Saúde
- Subjects
0301 basic medicine ,Life Cycles ,Cytoplasm ,Phagosomes / parasitology ,Vacuoles / drug effects ,Drug Evaluation, Preclinical ,lcsh:Medicine ,Protozoology ,Pattern Recognition, Automated ,White Blood Cells ,Macrophages / pathology ,Leishmania / drug effects ,Animal Cells ,Phagosomes ,Medicine and Health Sciences ,Macrophages / parasitology ,Femur ,Leishmaniasis / drug therapy ,lcsh:Science ,Leishmaniasis ,Cells, Cultured ,Protozoans ,Leishmania ,Staining ,Microscopy ,Mice, Inbred BALB C ,Vacuoles / pathology ,Multidisciplinary ,biology ,Eukaryota ,3. Good health ,Drug development ,Neglected tropical diseases ,Protozoan Life Cycles ,Leishmaniasis / diagnostic imaging ,Cellular Types ,Cellular Structures and Organelles ,Leishmania infantum ,Microscopy/methods ,Intracellular ,Research Article ,Amastigotes ,Imaging Techniques ,Immune Cells ,Leishmania Infantum ,Immunology ,Antiprotozoal Agents / pharmacology ,030106 microbiology ,Antiprotozoal Agents ,Macrophages / drug effects ,Vacuoles / parasitology ,Phagosomes / drug effects ,Image Analysis ,Computational biology ,Research and Analysis Methods ,Microbiology ,03 medical and health sciences ,Amphotericin B ,parasitic diseases ,Parasitic Diseases ,Animals ,Amastigote ,Cytoplasmic Staining ,Protocol (science) ,Blood Cells ,Science & Technology ,Tibia ,Macrophages ,lcsh:R ,Organisms ,Biology and Life Sciences ,Macrophage cell ,Cell Biology ,biology.organism_classification ,Parasitic Protozoans ,Pattern Recognition, Automated / methods ,Amphotericin B / pharmacology ,Phagosomes / pathology ,Specimen Preparation and Treatment ,Leishmania / cytology ,Drug Evaluation, Preclinical / methods ,Vacuoles ,lcsh:Q ,Software ,Developmental Biology - Abstract
Leishmania parasites cause a set of neglected tropical diseases with considerable public health impact, the leishmaniases, which are often fatal if left untreated. Since current treatments for the leishmaniases exhibit high toxicity, low efficacy and prohibitive prices, many laboratories throughout the world are engaged in research for the discovery of novel chemotherapeutics. This entails the necessity of screening large numbers of compounds against the clinically relevant form of the parasite, the obligatory intracellular amastigote, a procedure that in many laboratories is still carried out by manual inspection. To overcome this well-known bottleneck in Leishmania drug development, several studies have recently attempted to automate this process. Here we implemented an image-based high content triage assay for Leishmania which has the added advantages of using primary macrophages instead of macrophage cell lines and of enabling identification of active compounds against parasite species developing both in small individual phagolysosomes (such as L. infantum) and in large communal vacuoles (such as L. amazonensis). The automated image analysis protocol is made available for IN Cell Analyzer systems, and, importantly, also for the open-source CellProfiler software, in this way extending its implementation to any laboratory involved in drug development as well as in other aspects of Leishmania research requiring analysis of in vitro infected macrophages., Project Norte-01-0145-FEDER-000012Structured program on bioengineered therapies for infectiousdiseases and tissue regeneration, supportedby Norte Portugal Regional 17 Operational Programme (NORTE 2020), under the PORTUGAL 2020 PartnershipAgreement,through the European Regional Development Fund (FEDER). AGGA and TC were supported by contracts SFRH/BD/93766/2013,financed by POPH—QREN, and subsidized by Fundo Social Europeu and Ministe ´rio da Ciência,Tecnologia e Ensino Superior, and PTDC/QEQ-MED/7097/2014, funded by National Funds throughFCT, respectively. HC is fundedby FCT under the “Investigador FCT” Programme., info:eu-repo/semantics/publishedVersion
- Published
- 2018