Abstract
Optical diffraction tomography (ODT) provides label-free three-dimensional (3D) refractive index (RI) measurement of biological samples. However, due to the nature of the RI values of biological specimens, ODT has limited access to molecular specific information. Here, we present an optical setup combining ODT with three-channel 3D fluorescence microscopy, to enhance the molecular specificity of the 3D RI measurement. The 3D RI distribution and 3D deconvoluted fluorescence images of HeLa cells and NIH-3T3 cells are measured, and the cross-correlative analysis between RI and fluorescence of live cells are presented.
Highlights
The three-dimensional (3D) imaging of biological cells and tissues has been an invaluable tool for investigating subcellular organelles and various biological mechanisms [1]
Since optical diffraction tomography (ODT) and 3D deconvolution fluorescence microscopy employ the same optics setup, the 3D refractive index (RI) distribution and fluorescence images of cells are well-matched, which enables the identification of each subcellular component from the labelfree 3D RI distribution of the samples
The blue-channel fluorescence images clearly exhibit interior nucleus cells, which can be identified by a visual inspection of the 3D RI distribution of cells
Summary
The three-dimensional (3D) imaging of biological cells and tissues has been an invaluable tool for investigating subcellular organelles and various biological mechanisms [1]. Since the RI value of most biological specimen is linearly proportional to protein concentration [11, 12], ODT provides quantitative information including protein concentration and dry mass For these reasons, there are growing numbers of biological and medical studies employing ODT to investigate the physiology of various samples [3] including neuron cells [13, 14], red blood cells [15,16,17], parasites in host cells [18, 19], immune cells [20], bacteria [13, 21], gold nanoparticles in live cells [22], embryos [23], live cells in a microfluidic channel [24,25,26], and blood cells in vivo [27]
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
