Phagocytosis assays with different pH-sensitive fluorescent particles and various readouts.
Benjamin LindnerTobias BurkardMichael SchulerPublished in: BioTechniques (2020)
Phagocytosis is a fundamental mechanism of innate immunity and its impairment is associated with severe chronic diseases, for example, chronic obstructive pulmonary disease. Investigating phagocytosis requires flexible tools and assay conditions, such as different fluorescent particle types, detection colors and readouts. We comprehensively evaluated and optimized phagocytosis assays using particles labeled with fluorescent pH-sensitive pHrodo® dyes, facilitating the specific detection of phagocytosed particles. Beads, bacterial and yeast particles labeled with pHrodo red and green were tested for their uptake by THP-1 cells and primary human macrophages by flow cytometry and high-content imaging. Whereas the latter allowed kinetic phagocytosis measurement, the former demonstrated the feasibility of using cell sorting for periods of up to 6 h, enabling downstream applications such as pooled genetic screens.
Keyphrases
- label free
- high throughput
- flow cytometry
- chronic obstructive pulmonary disease
- quantum dots
- living cells
- genome wide
- endothelial cells
- induced apoptosis
- pet imaging
- high resolution
- loop mediated isothermal amplification
- randomized controlled trial
- gene expression
- cell cycle arrest
- dna methylation
- copy number
- computed tomography
- drug induced
- cell wall
- pet ct