3D correlative single-cell imaging utilizing fluorescence and refractive index tomography

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, quantifying the intracellular RI distribution and colocalizing it with fluorescence in 3D. 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.

15 pages, 5 figures

References in corpus (5)

• Structured illumination multimodal 3D-resolved quantitative phase and fluorescence sub-diffraction microscopy
• Refractive index tomography with structured illumination
• Diffraction microtomography with sample rotation: influence of a missing apple core in the recorded frequency space
• The Theory of Diffraction Tomography
• Refractive index measurements of single, spherical cells using digital holographic microscopy

Cited by in corpus (11)

• Quantitative phase imaging through an ultra-thin lensless fiber endoscope
• Accurate evaluation of size and refractive index for spherical objects in quantitative phase imaging
• Biobeam - Rigorous wave-optical simulations of light-sheet microscopy
• Integral refractive index imaging of flowing cell nuclei using quantitative phase microscopy combined with fluorescence microscopy
• Simultaneous off-axis multiplexed holography and regular fluorescence microscopy of biological cells
• Reconstructions of refractive index tomograms via discrete algebraic reconstruction technique
• Monitoring the refractive index of tissue models using light scattering spectroscopy
• Response to Comment on "Cell nuclei have lower refractive index and mass density than cytoplasm"
• Tailored 3D microphantoms: an essential tool for quantitative phase tomography analysis of organoids
• Imaging cloaked objects: diffraction tomography of realistic invisibility devices
• Wolf phase tomography (WPT) of transparent structures using partially coherent illumination