Automated high-content imaging for cellular uptake, from the Schmuck cation to the latest cyclic oligochalcogenides.
Rémi MartinentJavier López-AndariasDimitri MoreauYangyang ChengNaomi SakaiStefan MatilePublished in: Beilstein journal of organic chemistry (2020)
Recent progress with chemistry tools to deliver into living cells has seen a shift of attention from counterion-mediated uptake of cell-penetrating peptides (CPPs) and their mimics, particularly the Schmuck cation, toward thiol-mediated uptake with cell-penetrating poly(disulfide)s (CPDs) and cyclic oligochalcogenides (COCs), here exemplified by asparagusic acid. A persistent challenge in this evolution is the simultaneous and quantitative detection of cytosolic delivery and cytotoxicity in a high-throughput format. Here, we show that the combination of the HaloTag-based chloroalkane penetration assay (CAPA) with automated high-content (HC) microscopy can satisfy this need. The automated imaging of thousands of cells per condition in multiwell plates allows us to obtain quantitative data on not only the fluorescence intensity but also on the localization in a very short time. Quantitative and statistically relevant results can be obtained from dose-response curves of the targeted delivery to selected cells and the cytotoxicity in the same experiment, even with poorly optimized cellular systems.
Keyphrases
- high throughput
- high resolution
- single cell
- living cells
- induced apoptosis
- single molecule
- cell cycle arrest
- deep learning
- machine learning
- cell therapy
- ionic liquid
- fluorescent probe
- bone marrow
- mass spectrometry
- label free
- signaling pathway
- oxidative stress
- working memory
- big data
- cell death
- loop mediated isothermal amplification
- energy transfer
- real time pcr
- quantum dots