Three-dimensional correlative single-cell imaging utilizing fluorescence and refractive index tomography.
Mirjam SchürmannGheorghe CojocSalvatore GirardoElke UlbrichtJochen GuckPaul MüllerPublished in: Journal of biophotonics (2017)
Cells alter the path of light, a fact that leads to well-known aberrations in single cell or tissue imaging. Optical diffraction tomography (ODT) measures the biophysical property that causes these aberrations, the refractive index (RI). ODT is complementary to fluorescence imaging and does not require any markers. The present study introduces RI and fluorescence tomography with optofluidic rotation (RAFTOR) of suspended cells, facilitating the segmentation of the 3D-correlated RI and fluorescence data for a quantitative interpretation of the nuclear RI. The technique is validated with cell phantoms and used to confirm a lower nuclear RI for HL60 cells. Furthermore, the nuclear inversion of adult mouse photoreceptor cells is observed in the RI distribution. The applications shown confirm predictions of previous studies and illustrate the potential of RAFTOR to improve our understanding of cells and tissues.
Keyphrases
- induced apoptosis
- single cell
- cell cycle arrest
- fluorescence imaging
- high resolution
- cell death
- stem cells
- endoplasmic reticulum stress
- oxidative stress
- signaling pathway
- computed tomography
- single molecule
- deep learning
- risk assessment
- bone marrow
- dna methylation
- cell therapy
- big data
- pi k akt
- convolutional neural network
- crystal structure
- high speed
- childhood cancer