Correlative three-dimensional fluorescence and refractive index tomography: bridging the gap between molecular specificity and quantitative bioimaging

Kyoohyun Kim; Wei Sun Park; Sangchan Na; Sangbum Kim; Taehong Kim; Won Do Heo; YongKeun Park

doi:10.1364/BOE.8.005688

Correlative three-dimensional fluorescence and refractive index tomography: bridging the gap between molecular specificity and quantitative bioimaging

Biomed Opt Express. 2017 Nov 17;8(12):5688-5697. doi: 10.1364/BOE.8.005688. eCollection 2017 Dec 1.

Authors

Kyoohyun Kim^{1

2

3}, Wei Sun Park⁴, Sangchan Na⁵, Sangbum Kim⁵, Taehong Kim⁵, Won Do Heo^{4

6}, YongKeun Park^{1

2

5}

Affiliations

¹ Department of Physics, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 34141, South Korea.
² KI for Health Science and Technology (KIHST), KAIST, Daejeon 34141, South Korea.
³ Current address: Biotechnology Center, Technische Universität Dresden, 01307 Dresden, Germany.
⁴ Department of Biological Sciences, KAIST, Daejeon 34141, South Korea.
⁵ TomoCube Inc., Daejeon 34051, South Korea.
⁶ Center for Cognition and Sociality, Institute for Basic Science (IBS), Daejeon 34141, South Korea.

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.

Keywords: (180.2520) Fluorescence microscopy; (180.3170) Interference microscopy; (180.6900) Three-dimensional microscopy.