A Simple Technique for Simultaneously Quantifying Cell Death Index and Infection Index on in vitro Cell- Based Susceptibility Assays
Student Research in Translational Medicine,
Vol. 6 (2024),
24 January 2024
Abstract
Background and Aim: It is a common practice for biomedical researchers to evaluate Leishmania infection indexes on in vitro cell cultures to address biological questions or test the efficacy of novel anti- parasitic compounds. In Leishmania infections that parasitize macrophages, infection indexes are usually determined by visual inspection of the cells directly using Giemsa staining. However, the mechanism of action of treatment on Leishmania parasites in this method is not evaluated, therefore motivating the finding of analysis approaches that allow simultaneously quantifying Leishmania infection index and cell death on in vitro cell cultures.
Methods: We proposed a method for simultaneously evaluating parasite infection indexes and cell death indexes using fluorescent microscopy. To perform such an analysis, we used a fluorescent dye mix of ethidium bromide and acridine orange (EB/AO), which binds nucleic acids.
Results: EB/AO achieved simultaneous quantification of infection index and death index under normal growth and treatment conditions that were comparable to the conventional Giemsa staining method while overcoming the drawbacks of the current method.
Conclusion: The cell death index and Leishmania infection index can be simultaneously quantified using our method, which is less user- skill- dependent and faster than the general test.
- Cell Death
- Infection Index
- Leishmania
- Giemsa staining
- AO/EB staining
How to Cite
References
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