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
- optical coherence tomography
- cell fate
- cell death
- high speed
- high throughput
- oxidative stress
- respiratory syndrome coronavirus
- stem cells
- mass spectrometry
- deep learning
- machine learning
- coronavirus disease
- highly efficient
- dna methylation
- anti inflammatory
- fluorescence imaging
- label free
- aqueous solution