3 years ago

Three-dimensional correlative single-cell imaging utilizing fluorescence and refractive index tomography

Three-dimensional correlative single-cell imaging utilizing fluorescence and refractive index tomography
Paul Müller, Jochen Guck, Elke Ulbricht, Gheorghe Cojoc, Salvatore Girardo, Mirjam Schürmann
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. A tomographic imaging method is presented that performs colocalization of quantitative refractive index and fluorescence data to study single, suspended cells in 3D. The paper confirms recent studies by showing that HL60 cell nuclei have a lower refractive index than the cytoplasm in situ. In addition, the method is used to validate the nuclear 3D refractive index inversion in adult mouse retina photoreceptor cells.

Publisher URL: http://onlinelibrary.wiley.com/resolve/doi

DOI: 10.1002/jbio.201700145

You might also like
Never Miss Important Research

Researcher is an app designed by academics, for academics. Create a personalised feed in two minutes.
Choose from over 15,000 academics journals covering ten research areas then let Researcher deliver you papers tailored to your interests each day.

  • Download from Google Play
  • Download from App Store
  • Download from AppInChina

Researcher displays publicly available abstracts and doesn’t host any full article content. If the content is open access, we will direct clicks from the abstracts to the publisher website and display the PDF copy on our platform. Clicks to view the full text will be directed to the publisher website, where only users with subscriptions or access through their institution are able to view the full article.