Image-Based Quantification of Cell Debris as a Measure of Apoptosis.
Magnus ÖlanderNiklas HandinPer ArturssonPublished in: Analytical chemistry (2019)
Apoptosis is a controlled form of cell death that can be induced by various diseases and exogenous toxicants. Common apoptosis-detection methods rely on fluorescent markers, which necessitate the use of costly reagents and time-consuming labeling procedures. Label-free methods avoid these problems, but often require specialized instruments instead. Here, we utilize apoptotic-cell disintegration to develop a novel label-free detection method based on the quantification of subcellular debris particles in bright-field-microscopy images. Debris counts show strong correlations with fluorescence-based annexin V staining and can be used to study concentration-dependent and temporal apoptosis activation. The method is rapid, low-cost, and easy to apply, as the only experimental step comprises bright-field imaging of culture-media samples followed by automated image processing. The late-stage nature of the debris measurement means that the method can complement other, established apoptosis assays, and its accessibility will allow a wider community of researchers to study apoptotic cell death.
Keyphrases
- cell death
- label free
- cell cycle arrest
- endoplasmic reticulum stress
- deep learning
- oxidative stress
- low cost
- single cell
- healthcare
- cell therapy
- high resolution
- single molecule
- pi k akt
- high throughput
- loop mediated isothermal amplification
- quantum dots
- anti inflammatory
- palliative care
- signaling pathway
- convolutional neural network