Three‐dimensional correlative single‐cell imaging utilizing fluorescence and refractive index tomography. Issue 3 (25th September 2017)

• Record Type: Journal Article
• Authors: Schürmann, Mirjam; Cojoc, Gheorghe; Girardo, Salvatore; Ulbricht, Elke; Guck, Jochen; Müller, Paul
• Is Part Of: Journal of biophotonics. Volume 11:Issue 3(2018)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2017-09-25
• Subjects: cell nucleus -- flow cytometry -- optical diffraction tomography -- optical trapping -- refractive index -- retina cells -- single‐cell analysis -- tomography; Photonics -- Periodicals; Optical materials -- Periodicals; Optics -- Periodicals; Medical instruments and apparatus -- Periodicals; 621.3605
• DOI: 10.1002/jbio.201700145
• Languages: English
• ISSNs: 1864-063X

Abstract

Abstract : 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. Abstract : 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.