Monitoring correlates of SARS-CoV-2 infection in cell culture using a two-photon-active calcium-sensitive dye.
Domokos MáthéGergely SzalayLevente CseriZoltán KisBernadett PályiGábor FöldesNoémi KovácsAnna FülöpÁron SzepesiPolett HajdrikAttila CsomosÁkos ZsemberyKristóf KádárGergely KatonaZoltán MucsiBalázs József RózsaErvin KovácsPublished in: Cellular & molecular biology letters (2024)
We propose a new two-photon microscopy-based method combined with a cell-internalized sensor to quantify the level of SARS-CoV-2 infection. We optimized the applied dye concentrations to not interfere with viral fusion and viral replication events. The presented method ensured the proper monitoring of viral infection, replication, and cell fate. It also enabled distinguishing intracellular details of cell damage, such as vacuole and apoptotic body formation. Using clustering analysis, 2P microscopy calcium fluorescence images were suitable to distinguish two different viral variants in cell cultures. Cellular harm levels read out by calcium imaging were quantitatively related to the initial viral multiplicity of infection numbers. Thus, 2P quantitative calcium imaging might be used as a correlate of infection or a correlate of activity in cellular antiviral studies.
Keyphrases
- high resolution
- single cell
- sars cov
- single molecule
- cell therapy
- cell fate
- cell death
- optical coherence tomography
- respiratory syndrome coronavirus
- oxidative stress
- mass spectrometry
- gene expression
- machine learning
- mesenchymal stem cells
- reactive oxygen species
- dna methylation
- coronavirus disease
- label free
- data analysis
- monte carlo